Principle of Genetics PDF

Principle of Genetics
This eCourse Developed By

TNAU (ICAR)
Index
SN Lecture Page No
1. History of Genetic 1-9
2. Mendel’s laws of inheritance and exceptions to the laws. 10-28
3. Types of gene action 29-37
4. Multiple alleles 38-43
5. Multiple factor hypothesis. 44-47
6. Quantitative traits - Qualitative traits and differences between them 48-54
7. Cytoplasmic inheritance 55-60
8. Methods of inducing mutations and C l B technique. 61-66
9. Mutation - it’s characteristic features – 67-75
10. Regulation Gene expression 76-82
11. Lac operon and Fine structure of Gene. 83-98
12. Ultra structure of cell and cell organelles and their functions. 99-118
13. Study of chromosome structure, morphology, number and types - 119-135

Karyotype and Ideogram.
14. Mitosis and meiosis - their significance and differences between 136-151
them.
15. DNA and its structure, function, types, modes of replication and 152-166
repair.
16. RNA and its structure function and types. 167-178
17. Transcription – Translation - Genetic code and outline of protein 179-202

synthesis.
18. Crossing over – Factors affecting it 203-215
19. Linkage 216-224
20. Numerical chromosomal aberrations (Polyploidy) 225-246
21. Structural chromosomal aberrations. 247-251

Principles of Genetics
Lecture 1
01. HISTORY OF GENETICS
The history of genetics started with the work of the Augustinian friar Gregor Johann
Mendel. His work on pea plants, published in 1866, described what came to be known
as Mendelian Inheritance. In the centuries before—and for several decades after—Mendel's
work, a wide variety of theories of heredity proliferated.
1900 marked the "rediscovery of Mendel" by Hugo de Vries, Carl Correns and Erich von
Tschermak, and by 1915 the basic principles of Mendelian genetics had been applied to a
wide variety of organisms—most notably the fruit fly Drosophila melanogaster. Led
by Thomas Hunt Morgan and his fellow "drosophilists", geneticists developed
the Mendelian model, which was widely accepted by 1925. Alongside experimental work,
mathematicians developed the statistical framework of population genetics, bringing genetic
explanations into the study of evolution.
With the basic patterns of genetic inheritance established, many biologists turned to
investigations of the physical nature of the gene. In the 1940s and early 1950s, experiments
pointed to DNA as the portion of chromosomes (and perhaps other nucleoproteins) that held
genes. A focus on new model organisms such as viruses and bacteria, along with the
discovery of the double helical structure of DNA in 1953, marked the transition to the era
of molecular genetics.
In the following years, chemists developed techniques for sequencing both nucleic acids
and proteins, while others worked out the relationship between the two forms of biological
molecules: the genetic code. The regulation of gene expression became a central issue in the
1960s; by the 1970s gene expression could be controlled and manipulated through genetic
engineering. In the last decades of the 20th century, many biologists focused on large-scale
genetics projects, sequencing entire genomes.
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HISTORICAL DEVELOPMENTS IN GENETICS
Year Scientist Contribution
1485 L. da Vinci Recommended the use of lenses for viewing small

objects
1590 Z. Janssen and Produced the first operational microscope.

H. Janssen
1665 R. Hooke Introduced the term “ cell ” and described cork cells.
1668 F. Redi Disproved the theory of spontaneous generation of

maggots.
1672 Malphigi Classified the tissues.
1674 A.van Leeuwenhoek Improved lens system of microscope by grinding.
1682 N. Crew Described bladders and pores in wood and pith.
1694 J.R. Camerarius Conducted early experiments on pollination and

reported the existence of sex in plants.
1700 R. Linnaeus Classified the biological organisms.
1761 J.C. Kolreuter Hybridized various species of tobacco and

concluded that each parent contributed equally to
the characteristics of the progeny.
1779 C.F Wolff Founder of embryology.
1809 J.B. Lamarck Coined the word “ biology ” and stressed the
importance of cell in living organisms. He put
forth the theory of inheritance of acquired
characters.
1824 Dutrochet Showed that all plants and animals are composed
of cells.
1825 F.V. Raspail Developed the frozen-section technique and used

iodine for detection of starch.
1835 H. von Mohli Emphasized the importance of protoplasm and

described cell division.
1837 R. Brown Discovered the nucleus in cells of flowering plants.
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1838 M.J. Schleiden and Formulated the cell theory in plants and animals.
T. Schwann
1840 J.E. Purkinj Gave the term “ protoplasm ”.
1845 A. Donne Used photomicroscopy for the first time.
1846 K. Nageli Showed that plant cells arise from the division of pre-
existing cells.
1846 G.B. Amici Showed that egg in the ovary is stimulated to develop
into an embryo by the entrance of pollen tube.
1858 R. Virchow Showed that animal cells arise from the division of pre -
existing cells.
1859 C. Darwin Put forth the theory of natural selection.
1862 Kolliker Used the term “ cytoplasm ”for the living

material surrounding the nucleus.
1865 G. Mendel Developed the fundamental principles of heredity.
1870 W. His Invented the microtome.
1871 F. Meischer Isolated nucleic acids from pus cells.
1873 H. Fol Described spindle and astral rays.
1875 O. Hertwig Studied reproduction in sea urchins and concluded that

fertilization involves the union of sperm and egg
nucleus.
1875 E. Strasburger Discovered cell division in plants and gave the terms “
cytoplasm ” and “ nucleoplasm ”.
1879 W. Flemming Introduced the term “ chromatin ”.
1879 H. Fol Showed that only one sperm enters the egg during
fertilization.
1881 E.G. Balbiani Discovered giant chromosomes in salivary glands of

Drosophila.
1882 W. Flemming Coined the term “ mitosis ” .
1883 W. Rouse Proposed that chromosomes contain genes which are

the units of heredity.
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1885 A.F.W. Schimper Introduced the term “ plastids ”.
1888 Th. Boveri C oined the term “ centrosomes ”.
1888 W. Waldeyer Coined the term “ chromosomes ”.
1892 O. Hertwig Proposed the protoplasm theory of inheritance.
1892 J. Ruckert Described lamp brush chromosomes in oocytes of shark.
1892 W. Weisman Stated that chromosomes are the most important part of
the nucleus.
1892 Th. Boveri Described meiosis in Ascaris.
1898 C. Golgi Described the golgi apparatus in nerve cells.
1898 C. Benda Discovered mitochondria in spermatozoa and other cells.
1899 S. Altman Introduced the term “ nucleic acid ”.
1900 C.E. Correns, R e-discovered Mendel’s laws of inheritance.

H. de Vries and
E. Tschermak
1901 E. Strasburger Introduced the term “ plasmodesmata ”.
1902 C.E. McClung Identified sex chromosomes in bugs.

1902 H. de Vries Coined the term “ mutation ”.
1902 W.S. Sutton Proposed the chromosome theory of heredity and

Th. Boveri identified chromosomes as carriers of genetic material.
1903 W. Waldeyer Proved centromeres are the chromosomal regions with

which the spindle fibres become associated during
mitosis
1905 L.Cuenot Discovered lethal genes affecting coat colour in mice.
1905 J.B. Farmer and Coined the term “ meiosis ”.

J.E. Moore
1906 W. Bateson Coined the term “ Genetics ”and proposed the concept
of allele.
1906 W. Bateson and Discovered genetic linkage in sweet pea.

R.C. Punnet
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1906 W.L. Johannsen Coined the terms “gene”, “genotype” and “phenotype”.
1909 W. Bateson Coined the term “ epitasis ”.
1909 C. Correns Reported cytoplasmic inheritance in Mirabilis jalapa.
1909 F.A. Janssens Indicated that chiasmata are produced by exchanges

between non -sister chromatids of homologous
chromosomes.
1910 T.H. Morgan Studied crossing over and recombination in Drosophila

and coined the term “ crossing over ”.
1910 H. Nilsson-Ehle Proposed the multiple factor hypothesis.
1911 A.H. Sturtevant Constructed the first linkage map in Drosophila.
1912 Vejdovsky Coined the term “ chromonema ”.
1915 T.H. Morgan Correlated genetic studies with cytological studies. He

put forth the theory of linkage and studied sex linked
inheritance in Drosophila melanogaster.
1917 C.E. Allen Discovered sex determination in plants.
1921 F.G. Banting Isolated insulin.

C.H. Best
1922 C.B. Bridges Put forth the genic balance theory of sex determination.
1923 C.B. Bridges Discovered duplications, deletions and translocations in

chromosomes.
1923 Crew Reported complete reversal of sex in hens.
1924 A.F. Blakeslee and Studied trisomics in Jimson weed (Datura stromonium).
J. Belling
1924 R. Feulgen Described a test to confirm the presence of DNA.
1926 A.H. Sturtevant Discovered inversions in chromosomes.
1927 G.K. Karpechenko Synthesized Raphano brassica.
1927 H.J. Muller Induced mutations in Drosophila melanogaster by X-rays
1928 L.J. Stadler Induced mutations in maize and barley by X-rays.
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1928 F. Griffith Conducted experiments on transformations in Diplococcus

pneumonia.
1931 C. Stern Gave cytological proof for crossing over in Drosophila.

1931 H. Creighton and Gave cytological proof for crossing over in maize.
B. McClintock
1932 M. Knoll and Developed the electron microscope.

E. Ruska
1933 M. Rhodes Reported cytoplasmic male sterility in corn.
1935 F. Zernicke Developed the phase contrast microscope.
1935 R.B. Goldschmidt Coined the term “ phenocopy ”.
1939 R.A. Steinberg Induced mutations in Aspergillus sp. with chemicals.
1944 O.T. Avery, Explained the significance of DNA and proved that it is the
C.M. MacLeod genetic material.
and M. McCarty
1946 C. Auerbach and Induced mutations in Drosophila melanogaster using chemicals.

J.M. Robson
1946 E.S. McFadden, Synthesized Triticum spelta in the laboratory.
E.R. Sears and
H. Kihara
1948 K.R. Porter Described the endoplasmic reticulum.
1950 B. McClintock Discovered jumping genes in maize.
1951 A. Muntzing Synthesized Triticale.
1952 A.D. Hershey and Provided experimental proof of DNA as genetic material.
M.J. Chase
1953 Robinson and Observed ribosomes in plant cells.

Brown
1953 J.D. Watson, Proposed the double helix model for DNA molecule.
F.H.C. Crick and
M.H.F. Wilkins
1954 E.R. Sears Produced monosomic series of “Chinese Spring ” variety of

wheat.
1955 S. Benzer Described the fine structure of gene –Cistron, Recon and Muton.
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1955 C. DeDuve Coined the term “ lysosomes ”.
1955 G.E. Palade Observed ribosomes in animal cells.
1955 L. Pauling Studied the relationship between the structure of the DNA
molecule and protein synthesis.
1958 G.W. Beadle, Put forth the one gene – one enzyme hypothesis.
E.L. Tatum and
J. Lederberg
1958 F.H.C. Crick Explained the central dogma of molecular biology.
1958 M.S. Meselson Proved experimentally that DNA replicates by semi-

and F.W. Stahl conservative mechanism.
1959 A. Kornberg and Synthesized the DNA molecule in vitro.

S. Ochoa
1961 A.E. Jacob and Explained the genetic regulatory mechanism in protein
J. Monod synthesis – Operon concept.
1968 N.W. Nirenberg , Deciphered the genetic code and polynucleotide synthesis.
H.G. Khorana and
H. Holley
1968 Woodcock and Isolated DNA from chloroplasts.

Fernandez
1974 Clande, Re-discovered a number of cell organelles by electron

G.E. Palade and microscope.
C. DeDuve
1975 R. Dulbecco, Discovered the mechanism of reverse transcription – Teminism.

H. Temin and
D. Baltimore
1975 N. Borlaug Responsible for development of dwarf wheat and green

revolution.
1978 D. Nathans , Isolated restriction enzymes.

H.O. Smith and
W. Arber
1985 Potrykus Used electroporation technique for direct gene transfer in

plants.
1986 Helentzaris Developed the RFLP map in maize and tomato.
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1986 Ow Transferred and studied the expression of gene for enzyme

lucifersase (causes fire flies to glow) in tobacco cells.
1987 Fischoff Developed insect resistant transgenic tomato plants with Bt

gene.
1987 K.B. Mullis Developed polymerase chain reaction technique.
1988 Ouozzo Developed transgenic tobacco with CMV coat protein.
1991 Oeller Developed transgenic tomato with an antisense gene.
1992 Vasil Developed herbicide resistant transgenic wheat.
1993 Sharp Proposed the split gene concept.

Roberts
1993 Smith Studied site directed mutagenesis.
1994 Gilman and Rodbell Studied G proteins and their role in turning external signals
into action within cells.
1995 Lewis, Volard and Studied the role of genes in organ differentiation.
Wieschaus
1997 I. Wilmut Cloned sheep – Dolly.
1997 Prusiner Studied prions – Mad cow disease.
1998 Delta & Pine Co. Developed the terminator gene technology.
1998 Monsanto Co. Developed bollguard variety of cotton.
1998 T. Wakayama and Created the first cloned mice.

R. Yanagimachi
2000 Roslin Institute Created the first cloned pigs.
2001 Advanced Cell Birth of first cloned Asian ox called “Gaur”.

Technology
2002 Natl. Institute of Created the first cloned rabbit

Agronomic
Research, France.
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Lecture 2
MENDEL’S LAWS OF INHERITANCE AND EXCEPTIONS TO THE LAWS
History
The assertion that life can instantaneously arise from non living matter is called
spontaneous generation. Here are the critical experiments that busted the myth. Although today
we understand that living things arise from other living things, the idea of spontaneous
generation was entrenched in the minds of man throughout most of history. Spontaneous
generation is the belief that, on a daily basis, living things arise from non living material. This
debunked belief is not the same as abiogenesis, the study of how life on earth could have arisen
from inanimate matter billions of years ago.
Aristotle and Spontaneous Generation (383-322)
Aristotle was one of the first to record his conclusions on the possible routes to life. He
saw beings as arising in one of three ways, from sexual reproduction, asexual reproduction or
nonliving matter. According to Aristotle, it was readily observable that aphids arise from the dew
on plants, fleas from putrid matter, and mice from dirty hay; and this belief remained
unchallenged for more than two thousand years.
Francesco Redi’s Experiments (late 1600s)
Redi was and Italian physician and one of the first to formally challenge the doctrine of
spontaneous generation. Redi's question was simple, “Where do maggots come from?”
According to spontaneous generation, one would conclude that maggots came from rotting
food. Redi hypothesized that maggots came from flies and designed an experiment, elegant in
its simplicity, to challenge spontaneous generation.
Redi put meat into three separate jars:
Jar #1 he left open. He observed flies laying eggs on the meat and the eventual development
of maggots.
Jar #2 he covered with netting. Flies laid their eggs on the netting and maggots soon
appeared.
Jar #3 he sealed. Flies were not attracted to this jar and no maggots developed on the meat.
This seems to be a clear demonstration of life giving rise to life. Yet it took another two hundred
years for people to accept spontaneous generation as a fallacy.
Anthony van Leeuwenhoek’s “Animalcules” (1600-1700s)
Leeuwenhoek was a Dutch cloth merchant, and due to his trade, he frequently used
lenses to examine cloth. Rather than employing lenses made by others, he ground his own, and
the expertise that he gained through lens crafting combined with a curious mind eventually led
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to an interest in microscopy. During his life, Leeuwenhoek assembled more than 250
microscopes, some of which magnified objects 270 times. Through magnification, he discovered
presence of “micro” organisms - organisms so tiny that they were invisible to the naked eye. He
called these tiny living things “animalcules,” and was the first to describe many microbes and
microscopic structures, including bacteria, protozoans and human cells.
John Needham & Lazzaro Spallanzani (1700s)
The debate over spontaneous generation was reignited with Leeuwenhoek’s discovery
of animalcules and the observation that these tiny organisms would appear in collected
rainwater within a matter of days. John Needham and Lazzaro Spallazani both set out to
examine Leeuwenhoek's animalcules.
Needham’s Experiment
John Needham was a proponent of spontaneous generation, and his beliefs were
confirmed when, after boiling beef broth to kill all microbes, within the span of a few days,
cloudiness of the broth indicated the respawning of microscopic life.
Spallazani’s Experiment
Lazzaro Spallazani noted a flaw in Needham’s experiment. The containers holding
Needham’s beef broths had not been sealed upon boiling. So Spallazani modified Needham’s
experiment, boiling infusions, but immediately upon boiling he melted the necks of his glass
containers so that they were not open to the atmosphere. The microbes were killed and did not
reappear unless he broke the seal and again exposed the infusion to air.
Louis Pasteur (1800s)
Pasteur, a French scientist who made great contributions to our understanding of
microbiology and for whom the process of “pasteurization” is named, repeated experiments
similar to those of Spallazani’s and brought to light strong evidence that microbes arise from
other microbes, not spontaneously.
Pasteur’s Swan-Necked Flasks
Pasteur created unique glass flasks with unusual long, thin necks that pointed
downward. These “swan-necked” flasks allowed air into the container but did not allow particles
from the air to drift down into the body of the flask.
The End of Spontaneous Generation
After boiling his nutrient broths, Pasteur found that these swan-necked containers would
remain free of microbes until he either broke the necks of the flasks, allowing particles from the
air to drift in, or until he tilted the flask so that the liquid came in contact with dust that had
accumulated at the opening of the flask. It was these carefully controlled experiments of Pasteur
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that finally put to rest the debate over spontaneous generation.

Preformation theory (Swammerdam and Bonnet. 1720 1793)
Preformation theory proposes that the only male and female is responsible for heredity.
The male gamete consists of a miniature figure of man’s body called as homunculus which is
responsible for heredity. Epigenesis (C.f.wolf (1733-1794) and K.E. Von Baer (1792-1876) said
that the different organs and tissues of adult plant and animals developed from the uniform
embryonic tissue and not from mere growth expansion of the miniature homunculi present in
eggs / sperms. Von Baer proposed that they developed through a sequential modification of the
embryonic tissue. This concept is universally accepted.
Swammerdam (1637-1680), for example, thought that a tiny preformed frog occurred in
the animal hemisphere of the frog egg and that became simply larger by feeding on the food
stored in the vegetal hemisphere of the egg. Another biologist, Hartsoeker (1695) published a
figure showing a miniature man known as mankin or homunculus in the head of the human
spermatazoa. Such preformation theories had been supported by Leeuwenhoek (1632-1723),
Malpighi (1673), Reaumur, Bonnet (1720- 1793), Spallanzani (1729-1799) and other workers of
17th and early 18th centuries. With the development of improved microscopy and other
cytological techniques in 17th and 18th centuries, it became clear to biologists that neither the
egg nor the sperm contained a preformed individual but that each was a relatively uniform,
homogeneous mass of protoplasm.
Particulate Theory
A French biologist Maupertius in 1698-1759 discards the preformation theory and
forwarded the concept of biparental through many tiny particles. According to him both the
parents produce the semen, which composed of many tiny particles. The semen of both the
parents unite and the embryo formed each organ of the embryo was supposed to be formed by
two particles. Each of which came from each parent. In the year 1732-1806 J.C. Koelreuter was
the first person to get fertile hybrids by artificial crossing two species of tobacco and concluded
that the gametes were the physical basis of heredity.
Pangenesis
Charles Darwin proposed this theory. According to pangenesis that each organ of an
individual produces very small almost invisible identical copies of itself called gemmules or
pangenes. These gemmules from various parts collected into the blood stream of animals. The
blood transports the gemmules into the reproductive organ, which produce gametes. Male and
female gametes unite to form zygotes. When these gives rise to a new organism, the gemmules
of different parts of the body give rise to the same kind of organs, tissues and cells, which
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produced them in the parents.

Lamarckism
A French biologist Lamark (1774-1829) considered the inheritance of acquired characters
to be the most important, if not the sole, mechanism of evolutionary changes. According to
urgent need, use and disuse of organs, the modification thus acquired will be transmitted to their
off spring.
Germplasm theory August Weismanís (1834-1914)
Germplasm theory explains that body of individual consists of two distinct types of tissues,
(1) somatoplasm (2) germplasm. Somatoplasm consists of all body tissues, which do not
contribute to the sexual reproduction. The germplasm on the other hand produces gametes that
are the basis of heredity. It is only applied to animals and plants in which distinction between
soma and germ can be made. Weismannís famous experiment of cutting off the tail of mice for
22 generations and observing that the progeny still had tail of normal length, proved that the
somatoplasm is not responsible for transmission of characters.
Cell Theory(1838)
Schleiden and Schwann proposed cell theory 1838. They concluded that all plant and
animal tissues were made of cells. It was also postulated that cell is the functional unit of living
organism. In 1846 Negeli said that all cells originated from preexisting cells. Virchow 1853
elaborated this and referred it as cell linkage theory.
Mendelian concept of hereditary
The laws of inheritance were derived by Gregor Mendel, a 19th century monk
conducting hybridization experiments in garden peas (Pisum sativum). Between 1856 and 1863,
he cultivated and tested some 29,000 pea plants. From these experiments he deduced two
generalizations which later became known as Mendel's Laws of Heredity or Mendelian
inheritance. He described these laws in a two part paper, "Experiments on Plant Hybridization"
that he read to the Natural History Society of Bruno on February 8 and March 8, 1865, and
which was published in 1866.
Mendel's findings allowed other scientists to predict the expression of traits on the basis of
mathematical probabilities. A large contribution to Mendel's success can be traced to his
decision to start his crosses only with plants he demonstrated were true-breeding. He also
measured only absolute (binary) characteristics, such as color, shape, and position of the
offspring, rather than quantitative characteristics. He expressed his results numerically and
subjected them to statistical analysis. His method of data analysis and his large sample size
gave credibility to his data. He also had the foresight to follow several successive generations
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(f2, f3) of his pea plants and record their variations. Finally, he performed "test crosses" (back-
crossing descendants of the initial hybridization to the initial true-breeding lines) to reveal the
presence and proportion of recessive characters. Without his careful attention to procedure and
detail, Mendel's work could not have had the impact it made on the world of genetics.
Mendel's Laws
Mendel discovered that by crossing white flower and purple flower plants, the result was
not a hybrid offspring. Rather than being a mix of the two, the offspring was purple flowered. He
then conceived the idea of heredity units, which he called "factors", one which is a recessive
characteristic and the other dominant. Mendel said that factors, later called genes, normally
occur in pairs in ordinary body cells, yet segregate during the formation of sex cells. Each
member of the pair becomes part of the separate sex cell. The dominant gene, such as the
purple flower in Mendel's plants, will hide the recessive gene, the white flower. After Mendel
self-fertilized the F1 generation and obtained the 3:1 ratio, he correctly theorized that genes can
be paired in three different ways for each trait; AA, aa, and Aa. The capital A represents the
dominant factor and lowercase a represents the recessive.
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Mendel stated that each individual has two factors for each trait, one from each parent.
The two factors may or may not contain the same information. If the two factors are identical,
the individual is called homozygous for the trait. If the two factors have different information,
the individual is called heterozygous. The alternative forms of a factor are called alleles. The
genotype of an individual is made up of the many alleles it possesses. An individual's physical
appearance, or phenotype, is determined by its alleles as well as by its environment. An
individual possesses two alleles for each trait; one allele is given by the female parent and the
other by the male parent. They are passed on when an individual matures and produces
gametes: egg and sperm. When gametes form, the paired alleles separate randomly so that
each gamete receives a copy of one of the two alleles. The presence of an allele doesn't
promise that the trait will be expressed in the individual that possesses it. In heterozygous
individuals the only allele that is expressed is the dominant. The recessive allele is present but
its expression is hidden. Mendel summarized his findings in two laws; the Law of Segregation
and the Law of Independent Assortment.
Law of Segregation (The "First Law")
The Law of Segregation states that when any individual produces gametes, the copies of
a gene separate, so that each gamete receives only one copy. A gamete will receive one allele
or the other. The direct proof of this was later found when the process of meiosis came to be
known. In meiosis the paternal and maternal chromosomes get separated and the alleles with
the characters are segregated into two different gametes.
Law of Independent Assortment (The "Second Law")
The Law of Independent Assortment, also known as "Inheritance Law", states that
alleles of different genes assort independently of one another during gamete formation. While
Mendel's experiments with mixing one trait always resulted in a 3:1 ratio between dominant and
recessive phenotypes, his experiments with mixing two traits (dihybrid cross) showed 9:3:3:1
ratios. But the 9:3:3:1 table shows that each of the two genes are independently inherited with a
3:1 ratio. Mendel concluded that different traits are inherited independently of each other, so
that there is no relation, for example, between a cat's color and tail length. This is actually only
true for genes that are not linked to each other.
Independent assortment occurs during meiosis I in eukaryotic organisms, specifically
metaphase I of meiosis, to produce a gamete with a mixture of the organism's maternal and
paternal chromosomes. Along with chromosomal crossover, this process aids in increasing
genetic diversity by producing novel genetic combinations.
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In independent assortment the chromosomes that end up in a newly-formed gamete are

randomly sorted from all possible combinations of maternal and paternal chromosomes.
Because gametes end up with a random mix instead of a pre-defined "set" from either parent,
gametes are therefore considered assorted independently. As such, the gamete can end up
with any combination of paternal or maternal chromosomes. Any of the possible combinations of
gametes formed from maternal and paternal chromosomes will occur with equal frequency. For
human gametes, with 23 pairs of chromosomes, the number of possibilities is 2^23 or 8,388,608
possible combinations. The gametes will normally end up with 23 chromosomes, but the origin
of any particular one will be randomly selected from paternal or maternal chromosomes. This
contributes to the genetic variability of progeny.
Rediscovery of Mendelís work
Mendel's conclusions were largely ignored. Although they were not completely unknown
to biologists of the time, they were not seen as generally applicable, even by Mendel himself,
who thought they only applied to certain categories of species or traits. A major block to
understanding their significance was the importance attached by 19th century biologists to the
apparent blending of inherited traits in the overall appearance of the progeny, now known to be
due to multigene interactions, in contrast to the organ-specific binary characters studied by
Mendel. In 1900, however, his work was "re-discovered" by three European scientists, Hugo de
Vries, Carl Correns, and Erich von Tschermak. The exact nature of the "re-discovery" has been
somewhat debated: De Vries published first on the subject, mentioning Mendel in a footnote,
while Correns pointed out Mendel's priority after having read De Vries's paper and realizing that
he himself did not have priority. De Vries may not have acknowledged truthfully how much of his
knowledge of the laws came from his own work, or came only after reading Mendel's paper.
Later scholars have accused Von Tschermak of not truly understanding the results at all.
Regardless, the "re-discovery" made Mendelism an important but controversial theory. Its most
vigorous promoter in Europe was William Bateson, who coined the term "genetics", "gene", and
"allele" to describe many of its tenets.
The model of heredity was highly contested by other biologists because it implied that
heredity was discontinuous, in opposition to the apparently continuous variation observable for
many traits. Many biologists also dismissed the theory because they were not sure it would
apply to all species, and there seemed to be very few true Mendelian characters in nature.
However, later work by biologists and statisticians such as R.A. Fisher showed that if multiple
Mendelian factors were involved in the expression of an individual trait, they could produce the
diverse results observed. Thomas Hunt Morgan and his assistants later integrated the
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theoretical model of Mendel with the chromosome theory of inheritance, in which the
chromosomes of cells were thought to hold the actual hereditary material, and create what is
now known as classical genetics, which was extremely successful and cemented Mendel's
place in history.
Mendel's Laws of Inheritance
Mendel postulated three laws, which are now called after his name as Mendel’s laws of
heredity. These are:
1. Law of dominance and recessive
2. Law of segregation
3. Law of independent assortment
1. Law of Dominance
Definition: When two homozygous individuals with one or more sets of contrasting characters
are crossed, the characters that appear in the F1 hybrids are dominant characters and those do not
appear in F1 are recessive characters.
Law of dominance- If there are two alleles coding for the same trait and one is dominant it will
show up in the organism while the other won't
Explanation : The dominance and recessive of genes can be explained on the basis of
enzymatic functions of genes. The dominant genes - are capable of synthesizing active
polypeptides or proteins that form functional enzymes, whereas the recessive genes (mutant
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genes) code for incomplete or non-functional polypeptides. Therefore, the dominant genes
produce a specific phenotype while the recessive genes fail to do so. In the heterozygous
condition also the dominant gene is able to express itself, so that the heterozygous and
homozygous individuals have similar phenotype.
Critical appreciation of Law of Dominance
Scientists conducted cross-breeding experiments to find out the applicability of law of
dominance. The experiments were conducted by Correns on peas and maize, Tschermak on peas,
by De Vries on maize etc., by Bateson and his collaborators on a variety of organisms, by
Davenport on poultry, by Furst on rabbits, by Toyama on silk moth and by many others. These
scientists observed that a large number of characters in various organisms are related as dominant
and recessive.
Importance of law of dominance
The phenomenon of dominance is of practical importance as the harmful recessive
characters are masked by the normal dominant characters in the hybrids. In Human beings a
form of idiocy, diabetes, haemophilia etc. are recessive characters. A person hybrid for all these
characteristics appears perfectly normal. Thus harmful recessive genes can exist for several
generations without expressing themselves.
Exceptions to Law of Dominance is the Incomplete Dominance. After Mendel several cases
were recorded by scientists, where F1 hybrids exhibited a blending of characters of two parents. These
hybrids were found to be midway between the two parents. This is known as incomplete dominance or
blending inheritance. It means that two genes of the allelomorphic pair are not related as dominant
and recessive, but each of them expresses itself partially. As for example, in four-o'clock plant,
Mirabilis jalapa, when plants with red flowers (RR) are crossed with plants having white flowers (rr), the
hybrid F 1 plants (Rr) bear pink flowers. When these F 1 plants with pink flowers are self-pollinated
they develop red (RR), pink (Rr) and white (IT) flowered plants in the ratio of 1 : 2 : 1 (F 2
generation).
2. Law of Segregation (Purity of Gametes)
Explanation - The law of segregation states that when a pair of contrasting factors or genes or
allelomorphs are brought together in a heterozygote (hybrid) the two members of the allelic pair remain
together without being contaminated and when gametes are formed from the hybrid, the two separate out
from each other and only one enters each gamete.
Example - Pure tall plants are homozygous and, therefore/possess genes (factors) TT; similarly
dwarf possess genes tt. The tallness and dwarfness are two independent but contrasting factors or
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determiners. Pure tall plants produce gametes all of which possess gene T and dwarf plants t type
of gametes.
During cross fertilization gametes with T and t unite to produce hybrids of F1 generation. These
hybrids possess genotype Tt. It means F1 plants, though tall phenotypically, possess one gene for
tallness and one gene for dwarfness. Apparently, the tall and dwarf characters appear to have
become contaminated developing only tall character. But at the time of gamete formation, the genes T
(for tallness) and t (for dwarfness) separate and are passed on to separate gametes. As a result, two
types of gametes are produced from the heterozygote in equal numerosity. 50% of the gametes
possess gene T and other 50% possess gene t. Therefore, these gametes are either pure for tallness
or for dwarfness. (This is why the law of segregation is also described as Law of purity of gametes).
F1 Plants Tt X Tt
T t T t
Gametes unite at random and when gametes are numerous all possible combinations can
occur, with the result that tall and dwarf appear in the ratio of 3 :1. The results are often represented
by Punnett square as follows:
Critical appreciation of law of segregation

It has been confirmed by cytological studies that dominance or no dominance, the law of
segregation holds good to all cases. Its far reaching applicability has made it rare biological generalization.
RR have only gene for round
Rr, rR have gene for round and wrinkle
Rr have only wrinkeld gene
RR Rr
R r
Rr
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R r
R RR Rr
Rr
r Rr
Round, Wrinkled - 3:1 ratio
3. Law of Independent Assortment

Definition: The inheritance of more than one pair of characters (two pairs or more) is studied
simultaneously, the factors or genes for each pair of characters assort out independently of the other pairs.
Mendel formulated this law from the results of a dihybrid cross.
Explanation: The cross was made between plants having yellow and round cotyledons and plants
having green and wrinkled cotyledons.
The F 1 hybrids all had yellow and round seeds. When these F 1 plants were self fertilized they
produced four types of plants in the following proportion:
(i) Yellow and round 9
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(ii) Yellow and wrinkled 3

(iii) Green and round 3
(iv) Green and wrinkled 1
The above results indicate that yellow and green seeds appear in the ratio of 9 + 3 : 3 + 1 = 3 : 1.
Similarly, the round and wrinkled seeds appear in the ratio of 9 + 3 : 3 +1 = 12:4 or 3 :1. This indicates that
each of the two pairs of alternative characters viz. yellow-green cotyledon colour is inherited independent
of the round-wrinkled character of the cotyledons. It means at the time of gamete formation the factor for
yellow colour enters the gametes independent of R or r, i.e, gene Y can be passed on to the gametes either
with gene R or r.
Cytological explanation of the results: In the above experiment yellow and round characters are
dominant over green and wrinkled characters which can be represented as follows:
(i) gene for yellow colour of cotyledons Y
(ii) gene for green colour of cotyledons y
(iii) gene for round character of cotyledons R
(iv) gene for wrinkled character of colyledons r
Therefore, plants with yellow and round cotyledons will have their genotype YYRR and those with
green and wrinkled cotyledons will have a genotype yyrr. These plants will produce gametes with gene YR
and yr respectively. When these plants are cross pollinated, the union of these gametes will produce F1
hybrids with YyRr genes. When these produce gametes all the four genes have full freedom to assort
independently and, therefore, there are possibilities of four combinations in both male and female
gametes.
(i)RY (ii) Ry (iii) rY (iv) ry
This shows an excellent example of independent assortment. These gametes can unite at
random producing in all 16 different combinations of genes, but presenting four phenotypes in the ratio of
9: 3: 3: 1.
Dihybrid ratio : RR yy - Round, yellow seeded ; Rr yy - Wrinkled and greed seeded
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Test cross
F1 Rr Yy x rr yy (recessive)
1:1:1:1
Critical appreciation of law of Independent Assortment-
The law of independent assortment fails to have a universal applicability. Cytological studies have
revealed that only those allelomorphs assort independently during meiosis, which are located in different
homologous pairs of chromosomes. But, if the allelomorphs for different characters are present in the same
homologous pair of chromosomes, these are passed on to the same gamete. Law of independent
assortment does not apply to such cases.
BIOLOGICAL SIGNIFICANCE OF MENDEL'S LAWS
Mendel's work remained burried for about three decades, but after its rediscovery, the laws are
being used for the various branches of breeding. These are use for improving the varieties of fowls and their
eggs; in obtaining rust-resistant and disease-resistant varieties of grains. Various new breeds of horses and
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dogs are obtained by cross breeding experiments. The science of Eugenics is the outcome of Mendelism,
which deals with the betterment of human race.
Mendelian deviation
Mendelian deviations or exceptions or anomalies includes
1) Incomplete dominance
2) Codominance
3) Lethal genes etc.
1. Incomplete dominance
Mendel always observed complete dominance of one allele over the other for all the seven
characters, which he studied, in garden pea. Later on cases of incomplete dominance were
reported. For example, in four ëoí clock plant (Mirabilis jalapa) there are two types of flower viz.,
red and white. A cross between red and white flowered plants produced plants with intermediate
flower colour i.e. pink colour in F1 and a modified ratio of 1 red: 2 pink: 1 White in F2.
Parents Red flower x White flower
RR x rr
F1 Rr pink flower
F2 1 Red (Rr) : 2 Pink (RR) : 1 White (rr)
Incomplete dominance in flowers of Mirabilis jalapa
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2. Codominance
In case of codominance both alleles express their phenotypes in heterozygote greater
than an intermediate one. The example is AB blood group in human. The people who have
blood type AB are heterozygous exhibiting phenotypes for both the IA and IB alleles. In other
words, heterozygotes for codominant alleles are phenotypically similar to both parental types.
The main difference between codominance and incomplete dominance lies in the way in which
genes act. In case of codominance both alleles are active while in case of incomplete
dominance both alleles blend to make an intermediate one.
Codominance - both genes fully expressed

3. Lethal genes
Gene, which causes the death of its carrier when in homozygous condition is called lethal
gene. Mendel’s findings were based on equal survival of all genotypes. In normal segregation
ratio of 3:1 is modified into 2:1 ratio. Lethal genes have been reported in both animals as well as
plants. In mice allele for yellow coat colour is dominant over grey. When a cross is made
between yellow and grey a ratio of 1:1 for yellow and gray mice was observed. This indicated
that yellow mice are always heterozygous. Because yellow homozygotes are never born
because of homozygous
lethality. Such genes were not observed by Mendel. He always got 3:1 ratio in F2 for single
gene characters.
Lethal genes can be recessive, as in the aforementioned mouse experiments. Lethal
genes can also be dominant, conditional, semilethal, or synthetic, depending on the gene or
genes involved.
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MONOHYBRID CROSS
A cross is made between two true-breeding parents differing for a single trait, producing
an F1 generation. These plants are intercrossed to produce an F2 generation.
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Dihybrid Crosses
The following legends were described for peas by Mendel:
T- Tall
tt - dwarf
G - green (pod)
gg- yellow
Pure breeding parents can be crossed to produce a dihybrid meaning that 2 genes
affecting different traits are heterozygous (segregating) in all the f1 progeny.
Examples: TT, GG X tt, gg Tt, Gg
TT, gg X tt, GG Tt, Gg
When the F1 is self fertilized (plants) or crossed with another Tt, Gg individual, the
progeny will show the expected 3 dominant : 1 recessive phenotypic ratio for each trait. If the
two traits are independent, the two 3 : 1 ratios will interact to give a ratio based on 16ths.
# Genotypes Phenotypes
9 T_, G_ Tall, Green
3 T_, gg Tall, yellow
3 tt, G_ Dwarf, Green
1 tt, gg Dwarf, Yellow
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Backcross
Backcrossing is a crossing of a hybrid with one of its parents or an individual
genetically similar to its parent, in order to achieve offspring with a genetic identity which is
closer to that of the parent.
The Testcross
Because some alleles are dominant over others, the phenotype of an organism does not
always reflect its genotype. A recessive phenotype (yellow) is only expressed with the organism
is homozygous recessive (gg). A pea plant with green pods may be either homozygous
dominant (GG) or heterozygous (Gg). To determine whether an organism with a dominant
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phenotype (e.g. green pod color) is homozygous dominant or heterozygous, you use a
testcross.
The breeding of an organism of unknown genotype with a homozygous recessive. If all
the progeny of the testcross have green pods, then the green pod parent was probably
homozygous dominant since a GG x gg cross produces Gg progeny. If the progeny of the
testcross contains both green and yellow phenotypes, then the green pod parent was
heterozygous since a Gg x gg cross produces Gg and gg progeny in a 1:1 ratio. The testcross
was devised by Mendel and is still an important tool in genetic studies.
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Lecture 3
TYPES OF GENE ACTION

The interaction with in alleles of gene controlling a single character may be dominant,
incomplete dominance and co-dominance and are called intra allele interaction. When there is a
interaction occurs between different pairs of alleles influencing a character of an individual is
said to be interallelic interaction or epistatic. The gene that has masking effect is called epistatic
gene, and the gene whose effect is masked is known as hypostatic gene. Epistasis leads to
modification of normal dihybrid or trihybrid segregation ratio in F2 generation.
The term epistasis was coined by Bateson in 1909. Various types of epistatic gene
interaction are
1) Recessive epitasis (9:3:4)
2) Dominant epistasis (12:3:1)
3) Dominant and recessive (inhibitory) epistasis (13:3)
4) Duplicate recessive epistasis (9:7)
5) Duplicate dominant epistasis (15:1) and
6) Polymeric gene interaction (9:6:1).
Dihybrid ratio (9:3:3:1)
A classical case of two genes affecting the one and the same character and producing in
the F2 four different phenotypes in the ratio of 9:3:3:1 was discovered in fowls by Bateson and
Punnett. Each breed of poultry possesses characteristic type of comb. The Wyandotte breed
has a comb known as the rose comb, the Brahma has a pea comb, and the leghorn has a
single comb and the Malaya walnut comb. Each of these breeds true. Cross between rose
comb and single combed types show that rose in dominant to single comb and that there is a
segregation of 3 rose: 1 single comb in the F2. In mating between pea combed with single
combed and 3:1 ratio appears in F2. In mating between pea combed with single combed bird,
pea combed is found to be dominant over single comb and 3;1 ratio appears in F2. When a rose
combed fowl is crossed with a pea combed one, all the F1 birds show a new comb know as
walnut comb. When the walnut combs are inbred there appears in F2 walnut 3 rose pea single
comb. As well in the ratio of 9:3:3:1. The rose comb is due to the presence of R gene and Pea
due to P gene. Walnut comb is due to the presence of the dominant genes. R and P and single
comb are due to the presence of recessive of r and p. The ratio expected in F2 is 9:3:3:1.
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Parent RR PP x rrpp
Rose x rp
Rr pp(Walnut)
♂ RP Rp rP rp
♀
RP RRPP RRPp RrPP RrPp
(W) (W) (W) (W)
Rp RRPp RRpp RrPp Rrpp

(W) (R) (W) (R)
Rp RrPP RrPp rrPP rrpp

(W) (W) (P) (P)
Rp RrPp Rrpp rrPp rrPP

(W) (R) (P) (S)
9 Walnut: 3 Rose: 3 Pea: 1 Single
Duplicate recessive epistasis (Complimentary gene action) 9:7

When recessive alleles at either of the two loci can mask the expression of dominant
alleles at the two loci, it is called duplicate recessive epistasis. This is also known as
complementary epistasis. The best example of duplicate recessive epistasis if found for flower
colour in sweet pea. The purple colour of flower in sweet pea is governed by two dominant gene
say A and B when these genes are in separate individuals (Aabb or aaBB) and white (aabb)
they produce white flower. A cross between purple flower (AABB) and white flower (aabb)
strains produced purple colour in F1 intermating of F1 plants produced purple and white flower
plants in 9:7 ratio in F2 generation. Here the recessive allele .a. is epistatic to B/b alleles and
mask the expression of these alleles, another recessive allele b is epistatic to A/a alleles and
mask their expression.
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Parents purple x White

AABB x aabb
AB ab
AaBb
Purple
♂ AB Ab aB ab
♀
AB AABB AABb AaBB AaBb
(P) (P) (P) (P)
Ab AABb AAbb AaBb Aabb
(P) (W) (P) (W)
aB AaBB AaBb aaBB aaBb
(P) (P) (W) (W)
Ab AaBb Aabb aaBb aabb
(P) (W) (W) (W)
Ratio = 9 Purple : 7 white

Duplicate gene action (15:1) (Duplicate dominant epistasis)
When a dominant allele at either of two loci can mask the expression of recessive alleles
at the two loci, it is known as duplicate dominant epistasis. In rice awn character is controlled by
two dominant duplicate genes (A and B). Presence of any of these two alleles can produce awn.
The awnless condition develops only when both these genes are in homozygous recessive
state (aabb). A cross between awned and awnless strains produced awned plants in F1.
Intermating of F1 plants produced awned and awnless plants in 15:1 ratio in F2 generation. The
allele A is epistatic to a/b alleles and all plants having allele A will develop awn. Another
dominant allele B is epistatic to alleles a/b. An individual with these allele also develop awn
character.
Parents awned rice x awnless rice

AAbb x aaBB
AaBb
Awned rice
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♂ AB Ab aB ab
♀
(A) (A) (A) (A)
(A) (A) (A) (A)
(A) (A) (A) (A)
ab AABb AAbb AaBb Aabb
(A) (A) (A) (a)
Ratio = 15 awned : 1 awnless

Inhibitory gene action (13:3)
In this type of epistasis, a dominat allele at one locus can mask the expression of both
(dominant and recessive) alleles at second locus. This is also known as inhibitory gene
interaction. An example of this type of gene interaction is found for anthocyanin pigmentation in
rice. The green colour of plants is governed by the gene I which is dominant over purple colour.
The purple colour is controlled by a dominant gene P. when a cross was made between green
(IIpp) and (iiPP) colour plants, the F1 was green. Intimating of F1 plants produced green and
purple plants in 13:3 ratio in F2.
Parents awned rice x awnless rice
AAbb x aaBB
AaBb
Awned rice
♂ IP Ip iP ip
♀
IP IIPP (G) IIPp (G) IiPP (G) IiPP (G)
Ip IIPp (G) IIpp (G) IiPp (G) Iipp (G)
iP IiPP (G) IiPp (G) IiPP (P) iiPp (P)
ip IiPp (G) Iipp (G) iiPp (P) Iipp (G)
Ratio = 13 Green : 3 Purple
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Supplementary gene action. (Recessive epistasis) 9:3:4

Here one dominant gene has its own phenotypic effect and other dominant gene has no
effect of its own but its presence with the first gene modified the phenotypic expression. Thus in
supplementary gene action the dominant allele of one gene is necessary for the development of
the concerned phenotype, while the other gene modifies the expression of the first gene.
Parents RR PP x rr pp
Purple Red
RP rp
Rr Pp
Purple
♂ RP Rp RP Rp
♀
RP RRPP RRPp RrPP RrPP
(P) (P) (P) (P)
Rp RRPp RRpp RrPp Rrpp
(P) (W) (P) (W)
rP RrPp RrPp RrPP RrPp
(P) (P) (R) ®
rp RrPp Rrpp rrPp rrpp
(G) (W) (RP) (W)
Ratio = 9 Purple : 3 Red : 4 White
Additive factors (9:6:1) (Polymeric gene action)
In these two genes controlling a character produces identical phenotype when they are
alone i.e. with the homozygous recessive condition of the other gene. But when both the genes
are present together, their phenotype effect is enhanced as if the effect of the two genes were
cumulative or additives. It should be noted that in this case both the genes show complete
dominance. If the two genes showing polymeric gene action, what will be the consequence. In
barley two completely dominant genes A and B affect the length of awns, the thin needle like
extension of lemma genes A and B alone (e.g. Aabb and aaBB give gives rise to awn of
medium length, the effect of A is the same as that of B. But when both the genes A and B are
present together they produce long awn indicating the effect of A and B genes of awn length are
added together. Individual homozygous recessive for both these genes are awn less.
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Parents AA BB x aa bb
Long awned x awnless
Aa Bb
Long awned
♂ AB Ab aB ab
♀
AB AABB AABb AaBB AaBB
(L) (L) (L) (L)
(L) (A) (L) (A)
(L) (L) (A) (A)
ab AaBb Aabb aaBb aabb
(L) (A) (A) (a)
Ratio = 9 Long awned: 6 Awned: 1 awnless
11. Dominant Epistasis (12:3:1)

An example of dominant epistasis is found for fruit colour viz white,
yellow and green. White colour is controlled by dominant gene W and yellow colour by dominant
genes G. White is codominant over both yellow and green. The green fruits are produced in
recessive condition (wwgg). A cross between plants having white and yellow fruits produced F1
with white fruits. Intermating of F1 plants produced plants with white, yellow and green coloured
fruits in F2 was 12:3:1 ratio. Here W is dominant to w and epistatic to alleles G and g. Hence it
will mask the expression of G.g alleles. Hence in F2 plants with W-G- (9:16) and W-gg (3:16)
genotypes will produce white fruits; plants with wwG-3/16 will produce yellow fruits and those
with wwgg 1/16 genotype will produce green fruits. Thus the normal dihybrid ration 9:3:3:1 is
modified to 12:3:1 ratio in 1:2 generation. Similar type of gene interaction has been reported for
skin
colour in mice and seed coat colour in barley.
Parents White fruit x Yellow fruit

WWgg x wwGG
WwGg White fruit
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♂ WG Wg wG wg
♀
Wg WWGG WWGg WwGG WwGG
(W) (W) (W) (W)
Wg WWGg WWgg WwGg Wwgg

(W) (W) (W) (W)
WG WwGG WwGg wwGG WwGg

(W) (W) (Y) (Y)
Wg WwGg Wwgg wwGg wwgg

(W) (W) (Y) (G)
Ratio = 12 White: 3 Yellow: 1 Green
Modifying genes
These are group of genes, which enhances or reduce the phenotypic effect of a major
gene. Such genes have small and cumulative effect on the expression of the major genes. As a
result continuous variation is generated in the phenotype governed by a single major gene,
which converts qualitative character into a quantitative one. In rats, guinea pigs and rabbits,
piebald spotting is produced by recessive genes when present in a homozygous state (ss). The
degree of spotting depends upon the modifying factors, designed as S1, S2, S3 etc. which
enhances or reduces the expression of this spotting gene with cumulative on spotting. Most
quantitative characters of crop plants may be determined in a similar fashion. Some modifying
genes affect more than one character.
Major and minor genes
In the pie bald spotting the modifying factors produce some spotting even in the absence
of the spotting genes but their effect is much more pronounced in the presence of s, Obviously
the spotting gene s is a major gene controlling spotting, while the modifying genes are minor
genes affecting this trait.
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Inheritance of quantitative characters

Concept of polygenes
Colour, sex etc which shows distinct categories are known as qualitative characters.
They are usually governed by one or major genes or oligogenes. Characters like length of ear in
corn, yield of grain, yield of milk, stature etc do not fall into clear cut classes and shown more or
less continuous variation and are governed by a large number of minor genes called multiple
genes or polygenes. The characteristic feature of quantitative characters is 1) continuous
variation and 2) a marked influence of the environment on their expression.
Multiple factor hypothesis (Nilsson - Ehle 1908)
He effected crosses between different true breeding strains of wheat with red kernels
and with white kernel and the result of F1 and F2 were obtained. The F1 was (medium) red In
F2 15:1 ratio was obtained. Careful examination revealed that the red colour of F1 was not as
intene as the red colour of the parent and that in the F2 some red grains were as dark as those
of the parent and other only as dark as those of the F1. F2 plants revealed marked difference in
the intensity of colour depending upon the ratio of dominant and recessive genes present in
them. Thus he obtained 1:4:6:4:1 ratio for dark red, medium dark red medium red, light red and
white. It is evident that red colour is due to two pairs of genes. Each gene is capable of
producing red colour. Each is incompletely dominant over white and is cumulative in its effect.
The intensity of the red color depends upon the number of colour producing genes present.
Dark red is due the presence of four genes for red medium dark red to three genes, medium
red, to two genes and light red to one gene.
Nilson Ehle from his studies proposed the multiple gene hypothesis for the inheritance of
quantitative characters. This assumes that there is a series of independent genes for a given
quantitative traits. Dominance is usually incomplete and there is a strength of expression of the
character, whereas its alleles do not posses any effect. The F1 essentially uniform but
intermediate between the two parents. The F2 shows considerable variability, but is
intermediate between the two parents. The F2 mean value being approximately equal to the
parental mean and also the F1 mean. Studies on Nicotiana (East and Emerson 1916) worked
on quantitative characters to explain the inheritance of corolla length in nicotiana longiflora. He
crossed two inbred with contrasting corolla length of 40 and 93 mm. The F1 was intermediate
with mean corolla length of 63 mm. In F2 a much larger variation for corolla length than the
parents and F1 was observed. This variation was continuous as well and the F2 mean was
close to that of F1 and intermediate between those of the parent. This is precisely what is
expected in case of polygenic inheritance.
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Transgressive segregation
The appearance in F2 individuals with higher or lower intensity of characters than the
parents is called as transgressive segregation. It is produced when the parents have positive
alleles of different genes affecting a quantitative traits and segregation of these genes produce
two extreme homozygotes in F2, which transgress the parental limit for the character. The
reappearance of ancestor is called
atavism, throw back or reversion.
Expressivity
The degree of phenotypic expression of a penetrant gene is called expressivity. In other
words, the ability of a gene to produce identical phenotypes in all the individuals carrying it in
the appropriate genotype is known as incomplete expressivity. Many genes have incomplete
expressivity, while the wild type (normal) alleles are buffered against such variations.
Penetrance
The frequency with which a gene produces a phenotypic or visible effect in the
individuals, which carry it, is known as penetrance. In other words penetrance refers to the
proportion of individuals which exhibit phenotypic effect of a specific gene carried by them. In
general genes express themselves in all the individuals in which they are present in the
appropriate genotype is known as penetrance. It indicates the number of individuals that give
the expected phenotype to any degree.
Polydactyly
Polydactyly is a condition with extra fingers and toe or toes in man is due to the
presence of dominant gene P. The normal condition is produced by the genotype PP. The
genotype and pp produce polydactyly. Some heterozygous individual are not polydactly. (Pp).
Therefore the gene has penetrance of less than 100 per cent and said to be incompletely
penetrant. A gene though penetrant, may be quite variable in its expression. The degree of
expression produced by a penetrant genotype is termed expressivity. The polydactylous
condition may be penetrant in the left hand and not in the right hand or may be penetrant in the
feet and not in hands.
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Lecture 4
MULTIPLE ALLELES
Allele is a shorter term than allelomorph (another form) is the alternate form of gene.
Many genes have two alternate forms but several other have more than two alternate forms.
More than two alleles at the same locus give rise to a multiple allelic series. Multiple alleles can
be defined as a series of forms of a gene situated at the same locus of homologous
chromosomes. According to Mendel, each gene had two alternate forms or allele morphs are
being dominant and the other being recessive. Dominant being the wild type from which
recessive mutant was evolved through mutation. Likewise, a wild type can mutate in many ways
and produce many mutant forms and a mutant can again undergo another mutation and give
rise to a new mutant. Hence, a gene can exist in more than two allelomorphs. Usually wild type
allele is dominant over its recessive allele. wild allele is represented as + .
Multiple alleles can be defined as a
 series of forms of a gene
 situated at the same locus of homologous chromosomes
 affecting same character.
Multiple alleles are
 different forms of the same gene
 that is the sequence of the bases is slightly different in the genes located on the same
place of the chromosome.
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Multiple alleles are alternative states at the same locus. Remember: each individual will
only have two alleles for a trait but there are several alleles to choose from.) The classical
example for multiple alleles is human blood group self incompatibility in tobacco, coat colour in
rabbit, self incompatability genes in brassica.
The number of possible genotypes in a series of multiple alleles is ½ n (n+1)
n = no of alleles
 Di-allelic genes can generate 3 genotypes.
 Genes with 3 alleles can generate 6 genotypes.
 Genes with 4 alleles can generate 10 genotypes.
 Genes with 8 alleles can generate 36 genotypes
Important features of multiple alleles
1) Multiple alleles always belong to the same locus and one allele is present at a locus at a time
in a chromosome
2) Multiple alleles always control the same character of an individual
3) Wild type allele is dominant over other alleles
4) There is no crossing over in the multiple alleles
5) In a series of mutiple alleles wild type is always dominant
6) When two mutant types are crossed wild form cannot be recovered
7) The cross between two mutant alleles will always produce mutant phenotype. Examples of
multiple alleles are 1) fur colour in a rabbit, 2) ABO blood group in man 3) Wing type in
drosophila 4) Eye colour in drosophila etc. Fur colour in Rabbit. In rabbit, three alternate forms
of genes, which controls coat colour. C causes wild type and its alleles.
Skin colour in rabbit

In rabbits, four kinds of skin colour are known.
CC, Ccch, Cch, Cca - Agouti (wild type)

ch ch ch h ch
c ,c ,c c,c c - Chinchilla (salivary grey hair)
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ch ch, ch c - Himalayan (white except black feet nose ear tail)

cc - Albino (complete white).
Agouti
This has full colour and is also known as wild type. This colour is dominant over all the
remaining colour and produces agouti colour in F1 and 3:1 ratio in F2 when crossed with any of
the other three colours in rabbits. C represents this colour.
Chinchilla
This is lighter than agouti. This colour is dominant over Himalayan and albino and
produces chinchilla in F1 and 3:1 ratio in F2 when crossed either Himalayan or albino. This is
represented by cch.
Himalayan
The main body is white while the tips of ear, feet, tail and snout are coloured. This colour
is dominant over albino and produces 3:1 ratio in F2 when crossed with albino. This is
represented by ch .
Albino
This has pure white fur colour and is recessive to all other types. This is represented by
c. Thus the order of dominance for fur colour in rabbits can be represented as follows.
Agouti Chinchilla Himalayan Albino
(C) (cch) (ch) (c)
ABO Blood group in man.
Antibody
Antibody is a type of protein, which is commonly referred to as immunoglobin. It is
usually found in the serum or plasma. The presence of antibody can be demonstrated by its
specific reaction with an antigen.
Antigen
An antigen refers to an substance or agent, which when introduced into the system of
vertebrate animal like cow, goat, man etc induces the production of specific antibody, which
binds specifically to this (Antigen) substance Antigen are located in the red blood corpuscles
(RBC). If a person has a particular antigen in his RBCs, his serum has usually antibodies
against the other antigen. In human RBC two types of antigens viz A and B are present.
Depending upon the presence or absence of antigen A and B the blood group in man is of four
types viz A, B, AB and O. A person with blood group A has antigen A on the surface of RBCs:
protein with blood group B will have antigen B those with blood group AB have antigens A and
B; and those with blood group O have no antigen on the surface of their RBCs.
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Blood Genotype Antigen Antibody Compatible

Group found present blood group
A IAIA, IAIA A B A and O
B IBIB, IbIb B A B and O
AB IAIB AB None A,B, AB,O
O ii None AB O
Recent studies shows that antigen is galactosamine and B is galactose Antibodies A, B,

AB and None and are naturally present in the serum of individuals having A,B,AB, and O blood
group respectively. The agglutination or coagulation of RBCs leads to clotting of blood due to
interaction between antigen antibody. The blood group B cannot be transferred to an individual
having blood group A because the recipient has antibody against antigen B which is present on
the RBCs of blood group B. Similarly the reverse transfusion is not possible. The blood group
AB does not have antibody A and B. Hence individuals with AB blood group can accept all types
of blood, viz., A, B, AB and O. Such individuals are known as universal acceptors or
recipients. The O blood group does not have any antigen and has antibody against antigen A
and B, It cannot accept blood group other than O. Individuals with blood group O are known as
universal donors, because transfusion of blood group O is possible with all the four blood
types. The consideration of Rh (rhesus) type is important in blood transfusion. Each blood group
has generally two types of Rh group, viz positive and negative. The same type of Rh is
compatible for blood transfusion Opposite type lead to reaction resulting in death of the
recipient. These are few examples of multiple alleles Now it is believed that multiple alleles are
present almost for all genes.
Multiple alleles in plants

The classical example of multiple alleles in plants is ‘self incompatability alleles’ which
prevents self fertilization.
Multiple alleles in Maize

Multiple allelic series affecting seed color is seen in Maize.
A Mutation a Mutation a’
(purple) (white) light purple
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Multiple allele in cotton

L mutation l mutation lB ll mutant lI
narrow leaf Broad leaf mutant broad lacinated mutant intemediat
For information
 About 30% of the genes in humans are di-allelic, that is they exist in two forms.
 About 70% are mono-allelic, they only exist in one form and they show no variation.
 A very few are poly-allelic having more than two forms.
Pleiotropism
In general one gene affects a single character. But many genes are known to affect
more than one character such genes are known as pleiotropic genes and the condition is
termed as pleiotrophy. An example of a pleiotropic gene in human beingsis the recessive gene s
which produces sickle cell anemia in the ss homozygotes. These gene causes changes in two
or more parts of characters, which are not related, then the gene is said to be pleiotropic gene.
E.g. In cotton the Punjab hairy lintless gene lic produces seeds without lint. This gene also
causes incomplete lancinations of the leaf, reduction in boll size and fertility. In a plant a gene
may produce red pigment in several organs, such as flowers stem, leaves but still it is not
correct to say that the gene is pleiotropic because the gene has only one general effect, the
production of pigment. A gene for wing may be vestigial gene can be called as bristle gene or a
fecundity gene. A number of other recessive genes produce marked and often detrimental effect
in human beings. They are referred as syndromes.
Penetrance
Most genes produce identical phenotypes in all the individuals in which they are present
in the appropriate genotype. For example, all the seeds having the w gene governing the seed
shape in pea, in the homozygous state (ww) have uniformly wrinkled shape. Similarly, those
seeds that have either WW or Ww genotype are uniformly round. The ability of a gene to
produce identical phenotypes in all the individuals carrying it in the appropriate genotype is
known as complete expressivity. As opposed to this, many genes have incomplete expressivity
in that they produce variable phenotypes in the individuals that have this gene in the appropriate
genotype.
Expressivity
In general, genes express themselves in all the individuals in which theya re present in
the appropriate genotype, this is known as complete penetrance. But many genes do not
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produce the concerned phenotype in all the individuals which carry them in the appropriate
genotype. Such a situation is known as incomplete prenetrance. When a gene is present in the
appropriate genotype, the per cent of individuals in which it is able to express itself is a measure
of its penetrance. Thus the chlorophyll deficiency gene in lima beans has a penetrance of 10 %.
Almost all the genes showing incomplete penetrance exhibit incomplete expressivity as well.
Thus incomplete penetrance is in fact an expression of incomplete expressivity in that some
individuals show such a small expression of the gene that the trait is not detectable.
Isoalleles
These alleles, which are similar but on testing it proves to be a different one. Blood
group A person have three slightly different types such as IA1, IA2, IA3 which are similar but
found to be different after testing.
Pseudoalleles
The genes that are so closely linked can be separatable only by rare crossing over.
Such genes are called pseudoalleles.
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Lecture 5
MULTIPLE FACTOR HYPOTHESIS

Multiple factor
It is quite natural that small differences exist among individuals of similar genotype due
to the effect of environment on genotype. On the other hand, there are some heritable
differences also exist with continuous variation. Most of the economical traits show continuous
variation and they are measurable or quantifiable.
Quantitative characters
Quantitative characters are traits which show continuous variation and governed by a
large number of genes called multiple genes or multiple factors or polymeric genes or
polygenes. Their inheritance follows same mendelian principles.
Quantitative characters
Qualitative characters show
 discontinuous variation and
 are governed by one or two major genes or oligognes.
Multiple factor Hypothesis (Nilson – Ehle)

Nilson-Ehle studied Kernel colour in wheat concluded that is a quantitative character He
crossed true breeding red kernel whet (RR) with true breeding white (rr) and the F1 was red (Rr)
and the F2 segregated for red and white in 3:1 ratio indicating the dominance of red over white.
However, careful examination indicated the variation in red color among the red color progenies.
F1 red was not as intense as one of the parents. In F2 he could observe two grades of red ie.
one was red as that of one of its parent, two were higher red as that of F1 individuals. In some
crosses, a ratio of 15 red : 1 white was found in F2 indicating that there are two pairs of genes
for red colour that either or both of these can produce red kernels. Finally he observed different
shades of red in F2 for red kernel types. The F2 showed red shades and white as follows;
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Dark red : 1
Medium dark red : 4
Medium red : 6 15
Light red : 4
White : 1
Total : 16
 It was concluded two duplicate dominant alleles R1 and R2 cumulatively decide the
intensity of red colour
 and both R1 and R2 are in completely dominant over white.
 The high intensity of red colour depends on the number.
The F2 ratio in wheat
Genotype Genotypic ratio Phenotype
R1R1 R2R2 1 Dark red
R1R1 R2r2 2 Medium dark red
R1r1 R2R2 2 Medium dark red
R1r1 R2r2 4 Medium red
R1R1 r2r2 1 Medium red
r1r1 R2R2 1 Medium red
R1r1 r2r2 2 light red
r1r1 R2r2 2 light red
r1r2 r2r2 1 white
Hence, if two parents differ for the two genes the segregation was 1:4:6:4:1 provided
both R1 and R2 contribute equally to the colour. If three genes are involved in F2 segregation
showed 1:6:15:20:15:6:1 for red shades and 1 for white.
Thus, Nilson-Ehle’s multiple factor states that
i) For a given quantitative trait there could be several genes, which were independent in their
segregation, but had cumulative effect on phenotype
ii) Dominance is usually incomplete
iii) Each gene contributes something to the strength of expression of character whereas its
recessive allele does not of genes present dominance gene.
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Causes of Genetic Variation: The Multiple-factor Hypothesis

We are used to thinking about the genetics of discrete plant phenotypes, for example tall
versus short peas in Mendel’s experiments. In this example, variation (different alleles) at a
single gene contributes to a major phenotypic difference. However, for many traits of interest
(such as yield), it is often the case that a large number of genes, each of modest effect,
collectively contribute to the genetic variation. This is the multiple-factor hypothesis.
Early support for this multiple-factor hypothesis came from H. Nilsson-Ehle (1909), a
Swedish geneticist working on various cereal crops. Many of the characters that he examined
yielded 3:1 ratios in the F2 generation following the cross of two parental strains, consistent with
expectations for a single segregating locus with one allele completely dominant over the other.
However, there were some striking exceptions. For example, when red-seeded and white-
seeded wheat strains were crossed, the F1 progeny were identical in color (light red), but in
some of the F2 crosses, a ratio of 63 red:1 white seeds was observed. Nilsson-Ehle interpreted
this to be the result of the segregation of three independent factors, the initial parents being
AABBCC and aabbcc, all members of the F1 being AaBbCc and hence uniform in color, and the
F2 consisting of all possible genotypes, only one of which (aabbcc) gives rise to white seed. The
probability of obtaining an aabbcc offspring from an AaBbCc £ AaBbCc cross is (1=4)3 = 1=64:
From these results, Nilsson-Ehle arrived at two general conclusions. First, sexual reproduction
can produce a huge diversity of genotypes. For example, since a locus with two alleles A and a
can produce three genotypes (AA, Aa, and aa), ten diallelic loci can produce 310 ' 60; 000
genotypes. Second, given this huge potential diversity of genotypes, apparently new types
appearing within a population may be the result of rare segregants rather than new mutations.
Subsequent studies quickly confirmed these ideas. East (1911, 1916) and Emerson (1910;
Emerson and East 1913) examined quantitative variation in a large number of plants. Typically,
strains differing widely in some character were crossed and the variance of the resulting F1 and
F2 generations recorded. In most of these crosses, especially when the parental populations
were formed by repeated self-fertilizations, an outbreak of variation was seen in the F2 (Figure
1.2). Such outbreaks of variation, resulting from the segregation of multiple genotypes from the
F1 heterozygotes, are consistent with the multiple-factor hypothesis. For example, if the two
parents being crossed are inbred lines with genotypes AABBCC and aabbcc, then the resulting
F1 also has a single genotype AaBbCc. However, in the F2, all of the F1 heterozygotes can
segregate, so that an Aa parent can have AA, Aa, or aa offspring. Thus, the F2 consists of a
large collection of different genotypes, and hence is more variable (having both genetic and
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environmental contributions to variation) than either of the parental or F1 lines (which have only
environmental contributions to the variance).
Figure 1.2 The distribution of ear size in the F1 and F2 generations formed by crossing two
inbred lines of corn differing in ear length. The observed number of ears is given below each
size class. The variation seen in the P1, P2 and F1 populations is due entirely to environmental
factors, as all individuals in each population have the same genotype. These three populations
show roughly similar amounts of variation. In contrast, the F2 generation shows considerably
more variation, reflecting the diversity of genotypes in this population generated by segregation
of genes in the F1 parents. (Data from East 1911.).
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Lecture 6
QUANTITATIVE TRAITS - QUALITATIVE TRAITS AND DIFFERENCES BETWEEN THEM
Quantitative genetics (Inheritance of Multiple Genes)

The phenotypic traits of the different organisms may be of two kinds, viz., qualitative and
quantitative. The qualitative traits are the classical Mendelian traits of kinds such as form (e.g.,
round or wrinkle seeds of pea); structure (e.g., horned or hornless condition in cattles); pigments
(e.g., black or white coat of guinea pigs); and antigens and antibodies (e.g., blood group types
of man) and so on. We have already discussed in previous chapters that each qualitative trait
may be under genetic control of two or many alleles of a single gene with little or no
environmental modifications to obscure the gene effects. The organisms possessing qualitative
traits have distinct (separate) phenotypic classes and are said to exhibit discontinuous
variations.
The quantitative traits, however, are economically important measurable phenotypic
traits of degree such as height, weight, skin pigmentation, susceptibility to pathological diseases
or intelligence in man; amount of flowers, fruits, seeds, milk, meat or egg produced by plants or
animals, etc. The quantitative traits are also called metric traits. They do not show clear cut
differences between individuals and forms a spectrum of phenotypes which blend
imperceptively from one type to another to cause continuous variations. In contrast to qualitative
traits, the quantitative traits may be modified variously by the environmental conditions and are
usually governed by many factors or genes (perhaps 10 or I00 or more), each contributing such
a small amount of phenotype that their individual effects cannot be detected by Mendelian
methods but by only statistical methods.
Such genes which are non-allelic and effect the phenotype of a single quantitative trait,
are called polygenes or cumulative genes. The inheritance of poly genes or quantitative traits is
called quantitative inheritance, multiple factor inheritance, multiple gene inheritance or polygenic
inheritance. The genetical studies of qualitative traits are called qualitative genetics.
Certain Characteristics of Quantitative Inheritance
The quantitative inheritance have following characteristics:
1. The segregation phenomenon occurs at an indefinitely large number of gene loci.
2. If a substitution of a allele occurs in a gene locus then such allelic substitutions have
trivial effects.
3. The genes for a multiple trait have different biochemical functions but similar phenotypic
effects, therefore, the phenotypic effects of gene substitutions are interchangeable.
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4. Blocks of genes are bound together by inversions and transmitted as units from
inversion heterozygotes to their progeny, but such blocks are broken up by crossing over
in insersion homozygotes.
5. The polygenes have pleiotropic effects; that is, one gene may modify or suppress more
than one phenotypic trait. A single allele may do only one thing chemically but may
ultimately affect many characters.
6. The environmental conditions nave considerable effect the phenotypic expression of
poly genes for the quantitative traits. For example, height in many plants (e.g., corn,
tomato, pea, marigold) is genetically controlled quantitative trait, but some environmental
factors as soil, fertility, texture, and water, the temperature, the duration and wavelength
of incident light, the occurrence of parasites, etc., also affect the height. Similarly,
identical twins with identical genotypes, if grow up in different kinds of environments,
show different intelligence quotients.
Examples of Quantitative Inheritance
1. Kernel Colour in Wheat
Nilsson-Eble (1909) and East (1910, 1916) gave first significant clue of quantitative inheritance
by their individual works on wheat. They crossed a strain of red kernel wheat plant with another
strain of white kernel. Grain from the F 1 was uniformly red, but of a shade intermediate between
the red and white of the parental generation. This might suggest incomplete dominance, but
when F 1 offsprings were crossed among themselves, the F 2 zygotes showed five different
phenotypic classes in a. ratio of 1 : 4 : 6 : 4:1
Noting that 1/16 of the F 2 was an extreme in colour as either of the parental
plants (red or white), they theorized that two pairs of genes controlling production of red pigment
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while operating in this cross. Each gene was supposed to contain two alleles. One allele
produces a given quantity of the red pigment, while its counterpart did not produced any
pigment. All alleles were equally potent in the production or lack of production of pigment.
If we symbolize the genes for red with the capital letters A and B and their, alleles resulting in
lack of pigment production by a and b: We can illustrate the results of this cross as follows:or
1/16 Red: 4/16 Dark: 6/16 Medium: 4/16 Light: 1/16 White.
P: Red White
AABB x aabb
Gametes: (AB)(AB) (ab)(ab)
F1: Medium Medium
AaBb x AaBb
2.. Noting that 1/16 of the F 2 was an extreme in colour as either of the parental plants (red or
white), they theorized that two pairs of genes controlling production of red pigment while
operating in this cross. Each gene was supposed to contain two alleles. One allele produces a
given quantity of the red pigment, while its counterpart did not produced any pigment. All alleles
were equally potent in the production or lack of production of pigment.
If we symbolize the genes for red with the capital letters A and B and their, alleles resulting in
lack of pigment production by a and b: We can illustrate the results of this cross as follows:or
1/16 Red: 4/16 Dark: 6/16 Medium: 4/16 Light: 1/16 White.
P: Red White
AABB x aabb
Gametes: (AB)(AB) (ab)(ab)
F1: Medium Medium
AaBb x AaBb
♂ AB Ab aB ab
♀
Red Dark Dark Medium
Dark Medium Medium Light
Dark Medium Medium Light
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Ab AaBb Aabb aaBb Aabb

Medium Light Light White
3. Skin Colour in Man

Another classical example of polygenic inheritance was given by Davenport (1913) in
Jamaica. He found that two pairs of genes, A-a and B-b cause the difference in skin
pigmentation between Negro and Caucasian people. These genes were found to affect the
character in additive fashion. Thus, a true Negro has four dominant genes, AABB, and a white
has four recessive genes aabb. The F 1 offspring of mating of aabb with AABB, are all AaBb and
have an intermediate skin colour termed mulatto. A mating of two such mulattoes produces a
wide variety of skin colour in the offspring, ranging from skins as dark as the original Negro
parent to as white as the original white parent. The results of this cross are as follows :
Parents: Negro White
AABB x aabb
Gametes: ↓
F1: Mulatto Mulatto
AaBb x AaBb
↓
F 2 results:
Phenotypes Genotypes Genotypic Phenotypic Ratio
Frequency
Black (Negro) AABB 1 1
Dark AaBB, 2 4
AA Bb 2
Intermediate AaBb 4 6
aaBB 1
AA bb 1
Light Aabb 2 4
Aa Bb 2
White aa bb 1 1
These results are clearly showing that A and B genes produce about the same amount of
darkening of the skin ; and therefore, the increase or decrease of A and B genes cause variable
phenotypes in F 2 in the ratio of 1 Negro: 4 dark : 6 intermediate: 4 light : 1 white.
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4. Height in Man
Skin colour in man is a rather simple example of polygenic inheritance because only two
pairs of genes are involved. The inheritance of height in man is a more complex phenomenon
involving perhaps ten or more pairs of genes. The character of tallness is recessive to
shortness, thus, an individual having the genotype of more dominant genes will have the
phenotype of shortness. Because, this quantitative trait is controlled by multiple pairs of genes
and is variously influenced by a variety of environmental conditions. The heights of adults range
from 140 cm to 203 cm.
If one measured the height of a thousand adult men and the height of each is plotted against
height in centimeters and the points connected, a bell-shaped curve is produced which is called
curve of normal distribution and is characteristic of quantitative inheritance.
6. Other Examples
Likewise, if one measures the length of thousand sea shells the same species, or counts
the number of kernels per ear in a thousand ears of corn, or the number of pigs per litter in a
thousand litters, or weighs one thousand hen's egg, one will find a normal curve of distribution in
each case.
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Qualitative characters
The easiest characters, or traits, to deal with are those involving discontinuous, or
qualitative, differences that are governed by one or a few major genes. Many such inherited
differences exist, and they frequently have profound effects on plant value and utilization.
Examples are starchy versus sugary kernels (characteristic of field and sweet corn,
respectively) and determinant versus indeterminant habit of growth in green beans (determinant
varieties are adapted to mechanical harvesting). Such differences can be seen easily and
evaluated quickly, and the expression of the traits remains the same regardless of the
environment in which the plant grows. Traits of this type are termed highly heritable.
A qualitative trait is expressed qualitatively, which means that the phenotype falls into
different categories. These categories do not necessarily have a certain order. The pattern of
inheritance for a qualitative trait is typically monogenetic, which means that the trait is only
influenced by a single gene. Inherited diseases caused by single mutations are good examples
of qualitative traits. Another is blood type. The environment has very little influence on the
phenotype of these traits.
The major differences between the two are following:
Qualitative genetics Quantitative genetics
It deals with the inheritance of traits of kind, It deals with the inheritance of traits of degree,
viz., form, structure, colour, etc. viz., heights of length, weight, number, etc.
Discrete phenotypic classes occur which A spectrum of phenotypic classes occur which
display discontinuous variations contain continuous variations.
Each qualitative trait is governed by two or Each quantitative trait is governed by many
many alleles of a single gene. non-allelic genes or polygenes.
The phenotypic expression of a gene is not Environmental conditions effect the
influenced by environment. phenotypic expression of polygenes variously.
It concerns with individual matings and their It concerns with a population of organisms
progeny. consisting of all possible kinds of matings.
In it analysis is made by counts and ratios. In it analysis is made by statistical method
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Lecture 7
CYTOPLASMIC INHERITANCE
Inheritance of most of the characters in eukaryotic organisms shows the following

characteristic features.
1. The contributions by both male and female parents are equal so that the results from
reciprocal crosses are identical.
2. Segregation produces the characteristic 3:1 ratio in the F 2 generation of a
monohybrid cross and a typical 9:3:3:1in dihybrid crosses.
These features of inheritance were first demonstrated by Mendel: consequently,
such an inheritance pattern is referred to as Mendelian inheritance. It is universally
accepted that genes showing Mendelian inheritance are located in the chromosomes
of eukaryotic nuclei. Therefore Mendelian inheritance pattern is regarded as a
sufficient evidence for a gene to be located in the chromosomes, such genes are
termed as nuclear genes or more commonly simply as genes.
Non Mendelian Inheritance
But some characters in several organisms do not show Mendelian inheritance or they
show a non Mendelian inheritance pattern. In such cases, the following characteristic features
are observed.
1. There is consistent difference between the results from reciprocal crosses; generally
only the trait from female parent is transmitted.
2. In most cases, there is no segregation in the F2 and subsequent generations.
Characters showing non Mendelian inheritance may be grouped under three broad
categories:
(1) those related to cellular structures and patterns,
(2) those produced by intracellular parasites, symbionts and viruses
(3) those associated with DNA containing cell organelles viz., mitochondria and
chloroplasts.
In addition to these cases of non Mendelian inheritance, some characters in several organisms
exhibit a Mendelian inheritance pattern but the development of these characters in an individual
is markedly affected by the genotype of the maternal parent of the concerned individual; such
cases are classified as maternal effects.
The evidence for cytoplasmic inheritance was first presented by Correns in Mirabilis
jalapa and by Baur in Pelargonium zonale in 1908. In case of cytoplasmic inheritance generally
the character of only one of the two parents ( usually the female parent) is transmitted to the
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progeny. As a result, reciprocal crosses exhibit consistent differences for such characaters and
there is a lack of segregation in the F2 and the subsequent generations. Such inheritance is
also referred as extra nuclear inheritance, extrachromosomal inheritance and maternal
inheritance.
Genes governing the traits showing cytoplasmic inheritance are located outside the nucleus and
in the cytoplasm; hence they are referred to as plasma genes, cytoplasmic genes, cytogenes,
extranuclear genes or extra chromosomal genes.
The sum total of all the genes present in the cytoplasm of a cell is known as Plasmon,
while all the genes present in a plastid constitute a plastron.
Characteristics of cytoplasmic inheritance:
1. Reciprocal differences: Reciprocal crosses show marked differences for the characters
governed by plasmagenes. In most cases, plasmagenes from only one parent, generally the
female parent are transmitted, this phenomenon is known as uniparental inheritance.
2. Lack of segregation: In general, F2 F3 and the subsequent generations do not show
segregation for a cytoplasmically inherited trait. This is because the f1 individuals generally
receive plasma genes from one parent only.
3. Irregular segregation in biparental inheritance: In some cases, plasma genes from both
the parents are transmitted to the progeny, this is known as biparental inheritance.
4. Somatic segregation: Plasma genes generally show somatic segregation during mitosis, a
feature of rare occurrence in the case of nuclear genes.
5. Association with organelle DNA: Several plasma genes have been shown to be associated
with cp-DNA or mt-DNA.
6. Nuclear transplantation: If nuclear transplantation revealas a trait to be governed by the
genotype of cytoplasm and not by that of nucleus, cytoplasmic inheritance of the trait is strongly
indicated. In nuclear transplantation, nucleus of a cell is removed and replaced by a nucleus of
another genotype from a different cell. Generally nuclei of somatic cells are transplanted into
zygotes before the first mitotic division is initiated.
7. Transfer of nuclear genome through back crosses: The nucleus of a variety or species
may be transferred into the cytoplasm of another species or variety through repeated back
crossing with the former, which is used as the recurrent male parent. Lines produced in this way
are known as alloplasmic lines since they have nuclei and cytoplasms from two different
species. A comparison of the various characters of alloplasmic lines with those of the
corresponding euplasmic line (lines having nuclei and cytoplasms from the same species)
demonstrates cytopalsmic effects, if any on these traits. This technique is time consuming, but
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extremely powerful; it has been extensively used to study the cytopalsmic differentiation during
evolution.
8. Mutagenesis: Some mutagens eg: Ethidium bromide are highly specific mutagens for
plasma genes while nuclear genes are not affected by them.Induction of mutation by such
agenets in a gene indicates it to be a plasma gene.
9. Lack of chromosomal location: In many organism, extensive linkage maps of nuclear
genes are available. If a gene is shown to be located in one of these linkage groups, it cannot
be a plasma gene. Failure to demonstrate the location of a gene in one of the linkage groups of
an organism is indicative of its cytoplasmic location, but this is highly tentative.
10. Lack of association with a parasite, symbiont or virus: In many cases, a cytoplasmically
inherited character is associated with a parasite, symbiont or virus present in the cytoplasm of
the organism. Such cases cannot be regarded as cases of cytoplamic inheritance. Only those
cytoplasmically inherited characters which are not associated with parasites, symbionts or
viruses can be regarded as governed by plasma genes.
The known cases of true cytoplasmic inheritance are concerned with either choloroplast
or mitochondrial traits and are usually associated with their DNA. Such cases are therefore
often referred to as organellar inheritance, plastid inheritance and mitochondrial inheritance.
Plastid inheritance:
The inheritance pattern of plastid characters due to plasma genes located in plastid is
known as plastid inheritance. Plastid inheritance was first case of cytoplasmic inheritance to be
discovered independently by Correns and Baur in 1908. Variegation refers to the presence of
white or yellow spots of variable size on the green back ground of leaves. Variegation may be
produced by some environmental factors, some nuclear genes and in some cases, plasma
genes.
Inheritance of plastids in Mirabilis jalapa:
The inheritance of plastids in Four ‘O’ clock plant Meiabilis jalapa was first described by
Correns (1908). In M. Jalapa, some of the branches may have normal green leaves, while in the
same plant, some other branches may have only pale green or white leaves and still others may
have variegated leaves. Flowers on branches with normal green leaves produce seeds that
grow into plants with normal green leaves irrespective of whether they are pollinated by pollen
from branches with normal green variegated or pale green leaves.
Progeny of a variegated four ‘O’ clock plant
Type of branch from which Type of branch from which Type of leaf in the progeny
flowers are chosen for pollen was obtained grown from seed
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pollination
Green Green, Only green
variegated, “
pale green “
Variegated Green Green Green, variegated, or pale
Variegated “
Pale green “
Pale green Green Green, pale green
Variegated “
Pale green “
It is clear that variegation is determined by agencies transmitted through the female and
that it is not influenced by the type of pollen used. These agencies are the chloroplast. They are
capable of self-duplication and are transmitted from generation to generation through the
cytoplasm of the egg. Seeds borne on a green branch have three gene only green plastids,
seeds borne on a pale green branch have three gene only pale green plastids and seeds borne
on a variegated branch have green or pale green or a mixture of the two types of plastids.
Variegation is thus a heredity character determined by stable, self-duplicating, extra
nuclear particles called plastids. Neither the nucleus of the female gamete nor the male gamete
is involved in the control of this type of heredity character.
Maternal inheritance by ‘iojap’ gene in maize
The egg regularly contributes much more cytoplasm to the next generation than does
the sperm. It should therefore be expected that in cases of cytoplasmic inheritance, differences
between reciprocal crosses would result.
Rhoades (1946) identified the ‘iojap’ gene (ijij) in maize located in chromosome VII
controlling plastid inheritance in the plant. The gene ‘Ij’ is responsible for the normal green
colour of the plant.
When normal green plants with IjIj are used as female and pollinated by pollen from
stripped with ijij, F1 plants are wholly green.
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1. Green x stripped

Ij Ij ij ij
F1 Ijij Green
F2 3 green : 1 Iojap.
When striped with ijij are pollinated by pollen from the normal green plants with
IjIj the F1 plants, all of which have the same genotype. Ijij are of 3 different phenotypes.
Stripped ijij X Green Ij Ij

 (Ioja p) 
F1 Ijij Green, stripped or white (Iojap)
When plants with same genotype Ijij have different phenotype viz., normal green,
stripped or white, the differences can be attributed only the differences in plastids.
Cytoplasmic male sterility in Maize
In case of male sterility in maize, pollen grains of such male sterile are aborted. This
male sterility is transmitted only through the female and never by the pollen. When all of the
chromosomes of the male sterile line were replaced with chromosomes of normal plants, the
line still remained male sterile, showing thereby that male sterility in controlled by some agency
in the cytoplasm. It was later recognized that cytoplasmic male sterility in maize results from
alterations in the heredity units in the mitochondria (mitochondrial DNA).
Inheritance of Kappa particles in Paramecium
In Paramecium aurelia, two strains of individuals have been reported. One is called as ‘
Killer’ which secretes a toxic substance ‘ paramecin’ and the other strain in known as ‘ sensitive’
and is killed if comes in contact with the ‘paramecin’. In the cytoplasm of the killer strain the
kappa particles (cytoplasmic – DNA) are present kappa particles are absent in sensitive strains.
The transmission of kappa particles is through cytoplasm but maintenance of kappa particles
and production of paramecin is controlled by ‘k’ we assume that the killer strains carry dominant
allele ‘kk; and that sensitive ‘kk’. Conjugation Rare conjugation (cytoplasmic exchange)
On conjugation, congugents exchange their nuclear material so that ex-conjugants ‘kk’
resulted from conjugants ‘kk’ and ‘kk’ when conjugation is for normal time, then only nuclear
material is exchanged and therefore killer will produce killer daughters and sensitive will
produce sensitive daughters. But if the conjugation is in longer period, there will be exchange of
cytoplasm resulting in the inheritance of kappa particles by both the ex-conjugants so that all the
daughter paramecia produced are killers because all in herit the kappa particles through the
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mixing of cytoplasm. Therefore this trait is transmitted through cytoplasmic heredity. The trait is
only stable is killer strains.
Inheritance through mitochondria
Mitochondria can self-replicate and represent another genetic system in the cell. Of
course, the amount of mitochondrial DNA is so small, representing less than 1% of the nuclear
DNA is mammalian cells and it can code for a part of the protein in the mitochondria. The
synthesis of the cytochrome found in mitochondria for example, is known to be present in
minute amount in cytoplasm under the control of nuclear genes. Therefore, it is suggested that
both mitochondria and chloroplast seem to have a semiautonomous existence and their DNA
forms the basis for genetic systems separate from that in the nucleus.
Episome in Bacteria
Some hereditary particles have been found to exist in two states, either in an
autonomous state in the cytoplasm, where they replicate in dependently, of the chromosomes,
or in an integrated state incorporated into the chromosome. Particles with such properties are
known as episomes and include such things as the sex factor. The episomes are apparently not
essential to the life of the bacteria, because they may or may not be present. If they are absent,
they can be acquired only from an external source. In bacteria, E coli, sex is determined by the
presence or absence of the sex factor (F). Male bacterial cells (donor) have the sex factor and
this factor is responsible for the transfer of DNA from male to female bacterial cells (Recipient).
This sex factor is the cytoplasmic particle.
Significance of Cytoplasmic Inheritance
1. Development of cytoplasmic male sterility several crop plants like maize. Pearl millet,
sorghum, cotton etc.
2. Role of mitochondria in the manifestation of heterosis.
3. Mutation of chloroplast DNA and mitochondrial DNA leads to generation of new
variation
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Lecture 8
Methods of inducing mutations and C l B technique
Mutagens
Mutations can be induced by anumber of agents; the agents capable of
inducing mutations are called mutagens. Mutagen is a natural or human-made
agent (physical or chemical) which can alter the structure or sequence of DNA. The
different mutagenic agents may be classified into the following two broad groups:
1) Physical mutagens
2) Chemical mutagens
Physical mutagens
The different types of radiations having mutagenic properties are known as
physical mutagens. The radiations may be a part of the electromagnetic spectrum
having shorter wavelength and higher energy than visible light (eg: uv rays, X rays,
gamma rays and cosmic rays) or may be particulate radiations produced by the
decay of radio isotopes.
Radiations
Radiation was the first mutagenic agent known; its effects on genes were
first reported in the 1920's. Radiation itself was discovered in 1890's: Roentgen
discovered X-rays in 1895, Becquerel discovered radioactivity in 1896, and Marie
and Pierre Curie discovered radioactive elements in 1898. These three discoveries
and others led to the birth of atomic physics and our understanding of
electromagnetic radiation.
Radiations are grouped into two classes depending on the kind of effects they
have on the atoms in their path:
1. ionizing and
2. non – ionizing radiations
Non – ionizing radiations
Ultraviolet rays are the only non ionizing radiation with mutagenic properties.
The wave length ranges from 100 – 3900 Ao And they are specifically absorbed by
puriines and pyrimidines present in DNA. The maximum absorption of UV rays by
DNA as well as by pyrimidines, particularly thymine occurs at the wavelength of
254 nm, which is also the most mutagenic wavelength of UV.
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The mutagenic action of uv is the consequence of both its direct and indirect
effects on DNA. The direct effect of uv on DNA is of two types: formation of (1)
pyrimidine dimmers and pyrimidine hydrates.
Ionizing radiations
Ionizing radiations are so called because they cause ionization in the atoms
present in their path. There are two types of ionizing radiations: (1) particulate and
(2) non particulate radiations. Particulate radiations consist of high energy atomin
particles generated due to radioactive decay. The non particulate ionizing radiations
are represented by X rays and gamma rays which are high energy radiations
composed of photons.
The genetic effects of radiations may be (1) direct or (2) indirect. The direct
effect of radiations is produced due to ionizations directly in the DNA molecule,
while their indirect effect is produced through ionizations ini molecules other than
DNA and is believed to be mediated by free radical formation.
Sources of radiation
Natural sources of radiation produce so-called background radiation. These
include cosmic rays from the sun and outer space, radioactive elements in soil and
terrestrial products (wood, stone) and in the atmosphere (radon). One's exposure
due to background radiation varies with geographic location.
In addition, humans have created artificial sources of radiation which
contribute to our radiation exposure. Among these are medical testing (diagnostic
X-rays and other procedures), nuclear testing and power plants, and various other
products (TV's, smoke detectors, airport X-rays).
Chemical mutagens
The first report of mutagenic action of a chemical was in 1942 by Charlotte
Auerbach, who showed that nitrogen mustard (component of poisonous mustard
gas used in World Wars I and II) could cause mutations in cells. Since that time,
many other mutagenic chemicals have been identified and there is a huge industry
and government bureaucracy dedicated to finding them in food additives, industrial
wastes, etc. It is possible to distinguish chemical mutagens by their modes of
action; some of these cause mutations by mechanisms similar to those which arise
spontaneously while others are more like radiation in their effects.
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1. Base analogs
These chemicals structurally resemble purines and pyrimidines and may be
incorporated into DNA in place of the normal bases during DNA replication:
• bromouracil (BU)--artificially created compound extensively used in
research. Resembles thymine because it has Br in the 5 position instead of
methyl group and has the same effect on its base pairing behavior as that of
–CH 3 in the same position and therefore 5 BU behaves like thymine and
usually pairs with adenine.
• aminopurine --adenine analog which can pair with T or (less well) with C;
causes A:T to G:C or G:C to A:T transitions. Base analogs cause transitions,
as do spontaneous tautomerization events.
2. Chemicals which alter structure and pairing properties of bases
There are many such mutagens; some well-known examples are:
• nitrous acid--formed by digestion of nitrites (preservatives) in foods. It
causes C to U, meC to T, and A to hypoxanthine deaminations. Hypoxanthine
in DNA pairs with C and causes transitions. Deamination by nitrous acid, like
spontaneous deamination, causes transitions.
• nitrosoguanidine, methyl methanesulfonate, ethyl methanesulfonate-
chemical mutagens that react with bases and add methyl or ethyl groups.
Depending on the affected atom, the alkylated base may then degrade to
yield a baseless site, which is mutagenic and recombinogenic, or mispair to
result in mutations upon DNA replication.
3. Intercalating agents
Acridine orange, proflavin, ethidium bromide (used in labs as dyes and
mutagens). All are flat, multiple ring molecules which interact with bases of DNA
and insert between them. This insertion causes a "stretching" of the DNA duplex
and the DNA polymerase is "fooled" into inserting an extra base opposite an
intercalated molecule. The result is that intercalating agents cause frameshifts.
4. Agents altering DNA structure
This is used as a "catch-all" category which includes a variety of different kinds of
agents. These may be:
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• --large molecules which bind to bases in DNA and cause them to be

noncoding--we refer to these as "bulky" lesions (eg. NAAAF)
• --agents causing intra- and inter-strand crosslinks (eg. psoralens--found in
some vegetables and used in treatments of some skin conditions)
• --chemicals causing DNA strand breaks (eg. peroxides)
What these agents have in common is that they probably cause mutations not
directly but by induction of mutagenic repair processes.
Detection of mutation
The occurrence of mutational event at the gene level is detected by the

altercation it brings about in the phenotypic expression of one or more traits of the
concerned organism. Therefore the efficiency of detection of mutations will depend
largely on the availability of techniques for an easy and rapid scoring of the mutant
phenotypes in very large populations. Scoring of some types of mutations in certain
organisms is relatively easy. For example, mutations for antibiotic resistance in
bacteria are simply detected by plating the bacterial cells on a medium containing a
lethal concentration of the concerned antibiotic (selective medium); the colonies
that develop on such a medium will be produced by cells resistant to the antibiotic.
The medium lacking the antibiotic is called the non selective medium.
Number of colonies on the selective medium
Frequency of mutant cells (%) = --------------------------------------------------X 100
Number of colonies on the non selective medium
Detection of morphological mutations in eukaryotes requires examination of
each individual of the population for the mutant phenotype; this is not only tedious
requiring time, but is also a source of errors in the data. Therefore, elaborate
procedures for mutation detection have been developed in some eukaryotes . eg;
Drosophila, maize etc. These procedures employ specific markers to facilitate the
identification of chromosomes from the treated or irradiated individuals. Clearly
these techniques detect only germinal mutations. In drosophila, several specila
genetic stocks have been constructed for the detection of lethal and visible
mutations in X chromosomes and in autosomes; the two genetic stocks most
commonly used for mutation detection in X chromosome are (1) CIB and (2)
Attached X stocks.
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CIB Technique
This method was invented by Muller and used for the unequivocal
demonstration of mutagenic action of X rays. In this method, females containing
one normal X-chromosome and another X-chromosome (CIB) containing extra 3
genes are used for the analysis. Out of the 3 extra genes, one gene suppresses
crossover (c), the other is a recessive lethal (L) in heterozygous condition, and the
last gene is semidominant marker, Bar (B) gene.
Females containing CIB chromosome are called as CIB stock drosophila. The
normal males are exposed to mutagenic source for a fixed period and then mated
to the CIB stock drosophila. Males containing CIB chromosome will die due to the
effect of lethal genes, whereas norm ill males and females both normal and with
CIB will survive.
Females with CIB chromosomes and identified by barred phenotype are
selected and crossed to normal males. In this next generation 50% of males (which
have received the CIB gene) will die.
If mutation has occurred in normal X chromosome then even the normal
male (without CIB gene) will die. If no mutation has occurred all the other 50% of
males will survive. The frequency of lethal mutations can be accurately scored in
large samples. This technique is simple, rapid and there is little chance of an error
in scoring. However, it is suitable for the scoring of sex linked recessive lethal only.
The attached X chromosome technique
This technique is based on attached – X females (X- XY) and is designed to
study visible sex linked mutations in Drosophila. Mutagen treated males are mated
with attached X females. The X- XX (super female) and YY progeny produced from
such crosses do not survive; only X- XY (female) and XY (male) progeny are
recovered. All male (XY) progeny receive their Y chromosomes from their attached
X female parent, while their X chromosomes is contributed by their mutagen
treated male parent. If a visible mutation was induced in the X chromosomes of any
sperm produced by the mutagen treated male, it will be expressed in the male
progeny. Therefore all the male progeny obtained from the cross are scored for
visible mutations. The frequency of a visible mutation is expressed as the ratio
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between the number of progeny males showing a mutation and the total number of
males in the progeny.
Detection of mutations in plants
Techniques for the detection of mutations are relatively poorly developed for
plant species. The following two approaches are generally adopted for this purpose.
(1) In some species, eg; maize, strains homozygous for several recessive genes as
well as for dominant alleles of these genes are available. In such cases seeds or
plants of a strain homozygous for several dominant genes are treated with a
mutagen. The plants (M1 generation) are crossed with a strain having the recessive
forms of the traits governed by the concerned genes are counted and the frequency
of mutation for a gene is estimated as follows:
Number of plants having the recessive
form of trait governed by the gene
Mutation frequency (%) for a gene =-------------------------------------------------- X 100
Total number of plants in the progeny
The plants showing the recessive form of such a trait will receive one
recessive allele from the tester parent with the recessive traits, while the other
recessive allele would have been produced due to mutation in the mutagen treated
parent.
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Lecture 9
MUTATIONS
Introduction
Natures intention is that the exact genetic information from both
parents will be seen in the offspring's DNA in the critical stages of
fertilization. However, it is possible for this genetic information to mutate,
which in most cases, can result in fatal or negative consequences in the
outcome of the new organism.
Non-Disjunction and Down's Syndrome
One well known example of mutation is non-disjunction. Non-
disjunction is when the spindle fibres fail to separate during meiosis,
resulting in gametes with one extra chromosome and other gametes lacking
a chromosome. If this non-disjunction occurs in chromosome 21 of a human
egg cell, a condition called Down's syndrome occurs. This is because their
cells possess 47 chromosomes as opposed to the normal chromosome
compliment in humans of 46.
The fundamental structure of a chromosome is subject to mutation,
which will most likely occur during crossing over at meiosis. There are a
number of ways in which the chromosome structure can change, which will
detrimentally change the genotype and phenotype of the organism.
However, if the chromosome mutation effects an essential part of DNA, it is
possible that the mutation will abort the offspring before it has the chance of
being born.
Mutation
In most organisms genes are segments of DNA molecules. In the
broad sense, the term 'mutation' refers to all the heritable changes in the
genome, excluding those resulting from incorporation of genetic material
from other organisms. A mutation is an abrupt qualitative or quantitative
change in the genetic material of an organism. Mutations may be intragenic
or intergenic. Intragenic mutations or point mutations include alterations in
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the structure of the DNA molecule within a gene. In a point mutation there is
a change in the normal base sequence of the DNA molecule.
This change results in a modification of the structural characteristics or
enzymatic capacities of the individual. The unit of gene mutation is the
muton. This may consist of one or many nucleotide pairs. Intergenic
mutations, of which chromosomal changes in structure are examples,
involve long regions of DNA, i.e. many genes. These include deletion or
addition of segments of chromosomes, resulting in deficiency and
duplication, respectively. In large deletions a base sequence corresponding
to an entire polypeptide chain is sometimes lost. Such mutations are very
useful in genetic mapping.
Germinal and Somatic Mutations
Eukaryotic organisms have two primary cell types - germ and somatic.
Mutations can occur in either cell type. If a gene is altered in a germ cell, the
mutation is termed a germinal mutation. Because germ cells give rise to
gametes, some gametes will carry the mutation and it will be passed on to
the next generation when the individual successfully mates. Typically
germinal mutations are not expressed in the individual containing the
mutation.
Somatic cells give rise to all non-germline tissues. Mutations in
somatic cells are called somatic mutations. Because they do not occur in
cells that give rise to gametes, the mutation is not passed along to the next
generation by sexual means. To maintain this mutation, the individual
containing the mutation must be cloned. Two example of somatic clones are
navel oranges and red delicious apples.
Spontaneous and Induced Mutations
In general, the appearance of a new mutation is a rare event. Most
mutations that were originally studied occurred spontaneously. This class of
mutation is termed spontaneous mutations. But these mutations clearly
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represent only a small number of all possible mutations. To genetically

dissect a biological system further, new mutations were created by scientists
by treating an organism with a mutagenizing agent. These mutations are
called induced mutations.
The spontaneous mutation rate varies. Large gene provides a large
target and tends to mutate more frequently. A study of the five coat color
loci in mice showed that the rate of mutation ranged from 2 x 10-6 to 40 x
10-6 mutations per gamete per gene. Data from several studies on
eukaryotic organisms shows that in general the spontaneous mutation rate
is 2-12 x 10-6 mutations per gamete per gene.
Mutations can be induced by several methods. The three general
approaches used to generate mutations are radiation, chemical and
transposon insertion. The first induced mutations were created by treating
Drosophila with X-rays. In addition to X-rays, other types of radiation
treatments that have proven useful include gamma rays and fast neutron
bombardment. These treatments can induce point mutations (changes in a
single nucleotide) or deletions (loss of a chromosomal segment).
Other Types of mutations
Morphological mutants affect the outward appearance of an
individual. Plant height mutations could changes a tall plant to a short one,
or from having smooth to round seeds. Biochemical mutations have a
lesion in one specific step of an enzymatic pathway. For bacteria,
biochemical mutants need to be grown on a media supplemented with a
specific nutrient. Such mutants are called auxotrophs. Often though,
morphological mutants are the direct result of a mutation in a biochemical
pathway. In humans, albinism is the result of a mutation in the pathway
which converts the amino acid tyrosine to the skin pigment melanin.
Similarly, cretinism results when the tyrosine to thyroxine pathway is
mutated. For some mutations to be expressed, the individual needs to be
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placed in a specific environment. This is called the restrictive condition.

But if the individual grow in any other environment (permissive
condition), the wild type phenotype is expressed. These are called
conditional mutations. Mutations that only expressed at a specific
temperature (temperature sensitive mutants), usually elevated, can be
considered to be conditional mutations. Lethal mutations are also
possible. As the term implies, the mutations lead to the death of the
individual. Death does not have to occur immediately, it may take several
months or even years. But if the expected longevity of an individual is
significantly reduced, the mutation is considered a lethal mutation. If a
mutation occurs in that allele, the function for which it encodes is also lost.
The general term for these mutations is loss-of-function mutations. The
degree to which the function is lost can vary. If the function is entirely lost,
the mutation is called a null mutation. If is also possible that some function
may remain, but not at the level of the wild type allele. These are called
leaky mutations.
Types of Mutations
I. Chromosome Mutations - gross changes in chromosomes.
Changes in the number of chromosomes.
1. Euploidy - variation in the number of sets of chromosomes.
a. Haploidy (Monoploidy) - one set of chromosomes (n) : ABC
b. Polyploidy-three or more sets of chromosomes.
c. Triploidy-3 sets of chromosomes (3n) : ABC, ABC, ABC.
d. Tetraploidy-4 sets of chromosomes (4n): ABC, ABC, ABC, ABC.
e. Pentaploidy-5 sets of chromosomes (5n) : ABC, ABC, ABC, ABC, ABC.
f. Hexaploidy (6n), Septaploidy (7n), Octoploidy (8n), etc
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2. Aneuploidy - variation in the number of chromosomes of a set.

(Reduction in the normal number of chromosomes. )
a. Monosomics - Loss of one chromosome (2n-1) : ABC, AB.
b. Double monosomics - Ioss of 2 different chromosomes (2n-1-1): ABC,
A.
b - loss of a pair of homologous chromosomes (2n-2) : AB, AB:
b. Increase in the number of chromosomes (polysomies).
Trisomies - presence of 1 extra chromosome (2n+ 1) : ABC, ABC, A.
Double trisomics - 2 different extra chromosomes (2n + 1 + 1) : ABC,
ABC, AB.
Tetrasomics - an extra pair of homologous chromosomes (2n+2): ABC,
ABC, AA.
pentasomics (2n+3), Hexasomics (2n+4), Sepiasomics (2n+5), etc.
B. Changes in the structure of chomosomes.

a. Loss or addition of segments of chromosomes.
Deletion (deficiency) - Ioss of a segment of a chromosome
Duplication - repetition of a segment of a chromosome.
b. Changes in the normal arrangement of genes in the chromosome.
Translocation – Exchange of segments between two non - homologous
chromosomes, resulting in new chromosomes.
Inversion – Change in the linear order of genes by rotation of a section of a
chromosome through 180 degrees.
C. Gene mutations or point mutations – changes in the nucleotide

sequence of a gene.
a. Deletion
b. Insertion
c. Substitution
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d. Inversion
Rate of Mutation
The frequency of spontaneous mutations is usually low, ranging form
10-7 to 10-12 per organism. The rate of detectable mutations in average
gene is 1 in 106. It should be noted, however, that most methods for
estimating the rate of mutations tend to under estimate their frequency due
to many reasons. Firstly, lethal mutations which leave no progeny may be
missed. Secondly, mutations which leave only a slight change in the
phenotype may remain undetected. Mutations occur much more frequently
in certain regions of the gene than in others. The favoured regions are called
'hot spots'. Mutations involving single nucleotides can revert to normal gene
structure. Most single nucleotide mutations are reversible. In many cases
the rate of reverse mutations is similar to the rate of forward mutations. In
rare locations the rate of forward mutation is much greater than the rate of
backward mutation.
Effects of Mutations on the Phenotype
According to their effects on the phenotype mutations may be
classified as lethals, sub vitals and super vitals. Lethal mutations result in
the death of the cells or organisms in which they occur. Sub vital mutations
reduce the chances of survival of the organism in which they are found.
Super vital mutations on the other hand may result in the improvement of
biological fitness under certain conditions. There may also be mutations
which are neither harmful nor beneficial to the organism in which they occur.
How Does a Mutation Act?
As mentioned in other sections, genes act by controlling the rate of
production of specific proteins (enzymes). The scheme of protein synthesis
in most organisms is as follows:
(1) The DNA (gene) produces a complementary mRNA strand which has
codons consisting of nucleotide triplets.
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(2) tRNA molecules, each forming a complex with a specific amino acid, have
three free nuc1eotides which form the anticodon.
(3) The alignment of tRNA molecules on mRNA depends upon
complementary codon-anticodon pairing
(4) Thus the sequence of amino acid molecules in an enzyme (and hence the
structure and functions of the enzyme (depends upon the nucleotide
sequence of mRNA. This in turn depends upon the nucleotide sequence in
DNA. It will be seen that any change in the sequence of nucleotides of DNA
will result in a corresponding change in the nucleotide sequence of mRNA.
This may result in alignment of different tRNA molecules on mRNA. Thus the
amino acid sequence and hence the structure and properties of the enzyme
formed, will be changed. This may effect the traits controlled by the enzyme.
Molecular Basis of Mutations
Gene mutations at the molecular level involve modification of one base
by another, or addition or deletion of one or more bases. Mutations may be
spontaneous or induced Spontaneous Mutations - Mutations which occur
under natural conditions are called spontaneous mutations. It should be
noted that some spontaneous mutations arise by the action of mutagens
present in the environment. These mutagens include cosmic radiation,
radioactive compounds, heat, and such natural occurring base analogues like
caffeine. These will be considered under 'induced mutations' as they are
external agents bringing about mutations. Truly spontaneous mutations that
will be dealt with here are those arising from tautomerism.
Tautomerism
The ability of a molecule to exist in more than one chemical form is
called tautomerism .All the four common bases of DNA (adenine, guanine,
cytosine and thymine) have unusual tautomeric forms, which are, however,
rare. The normal bases of DNA are usually present in the keto form. As a
result of tautomeric rearrangement they can be momentarily transformed
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into the rare enol form in which the distribution of electrons is slightly
different. Normal base pairing in DNA is A-T and G-C. The tautomeric forms
are, however, capable of unusual ('forbidden') base pairing like T-G, G-T, C-
A and A-C. This unusual base pairing results in misreplication of the DNA
strand, giving rise to mutants in some of the progeny. Thus A*, a rare
tautomer of adenine (a) pairs with cytosine. This leads to G-C pairing in the
next generation. Spontaneous mutations can also arise as a result of
ambiguity of base pairing during replication because of 'wobble'
Induced Mutations A variety of agents increase the frequency of mutation.
Such agents are called mutagens. They include chemical mutagens, and
radiations like X-rays, γ-rays and UV-light.
Chemical Mutagens The first chemical mutagen discovered was mustard
'gas'. In the 1950s chemical mutagens with more or less specific action were
developed. Chemical mutagens can be classified according to the way in
which they bring about mutations:
(1) Base analogues which are incorporated into DNA instead of normal bases
(2) Agents modifying purines and pyridines and agents labilizing bases, and
(3) agents producing distortions in DNA.
The agents in categories (1) and (3) require replication for their action, while
agents in category (2) can modify even non replicating DNA.
Chemical mutagens work mostly by inducing point mutations. Two
major classes of chemical mutagens are routinely used. These are
alkylating agents and base analogs. Each has a specific effect on DNA.
Alkylating agents [such as ethyl methane sulphonate (EMS), ethyl ethane
sulphonate (EES) and musta rd gas] can mutate both replicating and non-
replicating DNA. By contrast, a base analog (5-bromouracil and 2-
aminopurine) only mutate DNA when the analog is incorporated into
replicating DNA. Each class of chemical mutagen has specific effects that can
lead to transitions, transversions or deletions.
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Scientists are now using the power of transposable elements to create

new mutations. Transposable elements are mobile pieces of DNA that can
move from one location in a geneome to another. Often when they move to
a new location, the result is a new mutant. The mutant arises because the
presence of a piece of DNA in a wild type gene disrupts the normal function
of that gene. As more and more is being learned about genes and genomes,
it is becoming apparent that transposable elements are a power source for
creating insertional mutants.
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Lecture 10
REGULATION OF GENE EXPRESSION
Each cell of a living organism contains thousands of genes. But all genes do
not function at a time. Genes function according to requirements of the cell. Genes
control the phenotypic expression of various characters through the production of
specific enzymes. Enzymes are special proteins which catalyse chemical reactions.
The production or synthesis of a particular enzyme is not constant. It varies as per
the requirement of the cell in other words, the synthesis of a particular enzyme is
sometimes high and sometimes low depending upon the requirement of the cell.
Thus, there exists an on-off system which regulates protein synthesis in all living
cells. The precise study of this on-off mechanism is called regulation of gene action
or regulation of gene expression or regulation of protein synthesis.
Synthesis of enzyme depends mainly on two factors. In a degradative

process, the synthesis of enzyme depends on the availability of the molecule to be
degraded. If the molecule is in more quantity, the enzyme synthesis will be more
and vice versa. In a biosynthetic pathway, the synthesis of an enzyme is governed
by the end product. If the end product is more, the enzyme synthesis will be less
and vice versa. There are two types of gene regulation, viz, (1) negative regulation,
and (2) positive regulation.
In negative regulation, this system and inhibitor is present in the cell, which
prevents transcription by inactivating the promoter. This inhibitor is known as
repressor. For initiation of transcription, an inducer in required. Inducer acts as
antagonist of the repressor. In the negative regulation, absence of product
increases the enzyme synthesis and presence of the product decreases the
synthesis.
Positive Control
In positive regulation, this system, an effectors molecule (which may be a
protein or a molecular complex) activates the promoter for transcription. In a
degradative system, either negative or positive mechanism may operate. In a
biosynthetic pathway negative mechanism usually operates.
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Important Terms
It is essential to define various terms which are commonly used in connection
with regulation of gene expression. A brief description of important terms is
presented below:
Brief description of important terms related to regulation of gene

expression
Terms Brief description
Repressor In operon, protein molecules which prevents transcription.

The process of inhibition of transcription is called
repression.
Inducer The substance which allows initiation of transcription (i.e.,

lactose in lac operon). Such process is known as induction.
Corepressor A combination of repressor and metabolite which prevents

protein synthesis. Such process is termed as corepression.
Inducible An enzyme whose production is enhanced by adding the

enzyme substrate in the culture medium. Such system is called
inducible system.
Repressible An enzyme whose production can be inhibited by adding

enzyme an end product. Such system is known as repressible
system.
Constitutive An enzyme whose production is constant irrespective of

enzyme metabolic state of the cell.
Negative control Inhibition of transcription by repressor through

inactivation of promoter e.g in lac operon.
Positive control Enhancement of transcription by an effector molecule

through activation of promoter.
Effector The molecule which acts as an inducer or corepressor in

the operon model of E.coli
The Operon Model
The operon refers to a group of closely linked genes which act together and
code for various enzymes of a particular biochemical pathway. In other words,
operon is a model which explains about the one-off mechanism of protein synthesis
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in a systematic manner. The operon model of gene regulation was proposed by

Jacob and Monod in 1961. They were awarded Nobel prize for this discovery in
1965. The operon model was developed working with lactose region (lac region) of
the human intestine bacteria E.coli. The gene regulation was studied for
degradation of the sugar lactose. The operon model consists of seven main
components, viz, (1) structural genes, (2) operator gene, (3) promotor gene, (4)
regulator gene, (5) repressor, (6) corepressor, and (7) inducer. A brief description
of these components is presented below:
Structural Genes
The lac operon of E.coli consists of three structural genes, viz, z, y and a.
The z gene is located near to the operator gene, y is located between z and a, and
a is located on right end of the operon segment. These structural genes transcribe a
single polycistronic mRNA molecule. This mRNA molecule controls the synthesis of
three different enzymes, viz., β- galacto-sidase, galactosidase permease and
galactosidase transacetylase. The enzyme galactosidase consists of 4 units and
catalyses the breakdown of lactose into glucose and galactose as given below:
The enzyme galactosidase permease is made up of one unit and permits

entry of lactose from the medium into the bacterial cell. Galactosidase acetylase
consists of two units. Its main function is to transfer an acetyl group from acetyl co-
enzyme A to β- galactosidase. The function of all the structural genes is controlled
by operator gene. Thus, the main function function of structural genes is to control
synthesis of protein through messenger RNA. In an operon, number of structural
genes is always equal to the number of polypeptide chains synthesized under
common control. If three types of proteins are synthesized from one operon, there
should be three structural genes. In prokaryotes, all the structural genes form
single polycistronic m RNA molecule, whereas in eukaryotes, each structural gene
forms separate (monocistronic) mRNA molecule.
Operator Gene
In lac operon of E. coli, the operator gene is located just near the structural
gene z. It consists of 35 nucleotide base pairs. It is the binding site for the
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repressor. The main function of operator gene is to control the function of structural
genes. However, its own function depends on the repressor molecule. Binding of
repressor with operator makes it non-functional and thus prevents transcription.
Repressor prevents transcription by inactivating the promoter gene. Mutation of
operator makes it unfit for binding with repressor. In such situations, operator is
free from binding with repressor and transcription can start. When the repressor is
bound to the operator, initiation of transcription of lac mRNA by RNA polymerase is
prevented. When operator is free, the promoter is available for initiation of mRNA
synthesis.
Promoter Gene
In lac operon of E. coli, the promotor gene is located next to operator. This
is located between operator gene and regulator gene. The promotor segment is a
place where mRNA polymerase enzyme binds with DNA. The recent investigations
(Pribnow, 19715) suggested that promotor segment has three sub regions, viz, (1)
a recognition site, (2) a binding site, and (3) an m RNA initiation site. The main
function of promotor gene is to initiate mRNA transcription. The m RNA
transcription moves from promotor region to the structural genes through operator
region. The promotor starts mRNA transcription only when operator is free or when
repressor is not bound to the operator gene. The binding of repressor with operator
inactivates the promotor gene and prevents transcription.
Regulator Gene
The regulator gene is located on one end of operon segment in E. coil. The
function of the regulator gene is to direct the synthesis of a repressor which is a
protein molecule. The repressor may be either active or inactive (a prorepressor).
Active repressor has a tendency to bind with operator gene in the inducible system.
The repressor binds to the operator in the absence of an inducer and prevents
mRNA transcription by inactivating the promoter gene. When an inducer (i.e.,
lactose) is present, the repressor binds to the inducer and forms an inducer –
repressor complex. This complex cannot bind to operator gene and, therefore,
protein synthesis can take place. In the repressible system, the repressor molecule
is inactive and, therefore, cannot bind with the operator gene. In such condition,
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protein synthesis by structural genes can take place. The repressor can become
active on combining with co-repressor. This repressor corepressor complex blocks
the operator gene and prevents protein synthesis.
Repressor
Repressor is a protein molecule. Its synthesis is directed by regulator gene.
It may be either in the active form or inactive form as described above. It has
affinity with operator gene. In the active form, it binds with operator gene and
prevents transcription and protein synthesis by inactivating promoter gene. When it
is in inactive form, the transcription and protein synthesis can take place. This can
be inactivated by an inducer.
Corepressor
Corepressor is perhaps a product of one of the enzymes synthesized by
structural genes. The corepressor makes the inactive repressor active in a
repressible system after combining with the same. The repressor – corepressor
complex can block the operator gene and stop protein synthesis by structural
genes.
Inducer
Inducer is a substrate (i.e. lactose in lac operon) which promotes
transcription. It binds with repressor molecule and makes the same inactive. The
repressor then cannot bind with operator gene. Hence, the transcription and protein
synthesis can take place.
Mechanism of Gene Regulation

The mechanism of gene regulation is of two types, viz, (1) negative
regulation, (2) positive regulation. These are briefly described below:
Negative Control
In the negative regulation, absence of a product enhances the synthesis of
enzyme and presence of the product decreases the synthesis of enzyme. In the lac
operon of E. coli. The synthesis of protein depends whether the operator gene is
blocked or free. When the operator gene is free, protein synthesis by structural
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genes will take place. On the other hand, when the operator gene is blocked, the
protein synthesis is prevented. Thus, the on-off of protein synthesis is governed by
the free and occupied position of the operator gene. In negative control, regulator
protein acts as a inhibitor and prevents protein synthesis. In lac operon of E.coli,
there is negative control of gene regulation. In the negative control, the regulator
protein is the repressor which inhibits protein synthesis. In the inducible system,
the effector molecule is the inducer. The inducer binds with repressor and
inactivates it so that it cannot bind with operator. Thus, inducer permits protein
synthesis by inactivating the repressor. In the repressible system, the effector
molecule is the corepressor. The corepressor on binding with in-active repressor
makes it active and inhibits protein synthesis, because when repressor becomes
active it will bind with operator and stop transcription.
Positive Control
In positive regulation, presence of a product will enhance the synthesis of
enzyme. In other words, in positive control the regulator protein acts as an
activator and enhances the protein synthesis. The arabinose operon of E.Coli is an
example of positive gene regulation.
Mutation of the Operon

The mechanism of gene regulation is affected by the mutation of genes in the
operon segment of E. Coli. The mutation of regulator gene will lead to production of
inactive or non-functional repressor. Such repressor is unable to bind with promotor
and, therefore, cannot inhibit transcription. In other words, defective repressor
cannot control the function of operator gene. In such situations, there is constant
synthesis of enzymes by the structural genes. This type of mutants is known as
constitutive mutants and the enzymes which are produced as a result of
constitutive mutations are referred to as constitutive enzymes.
Similarly, a mutation in the promotor region also alters the mechanism of

gene regulation. It may lead to three different situations depending upon the type
of mutation which takes place in the promotor gene.
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1. Total inactivation of promotor gene, as a result of mutation, will prevent

binding of RNA polymerase to the DNA at the promotor site.
2. An up-promotor mutation will permit rapid binding of RNA polymerase to the
promotor site and lead to enhanced transcription and enzyme sythensis.
3. A down – promotor mutant will decrease the rate of RNA polymerase binding
to promotor site and lead to reduction in enzyme synthesis.
Thus, mutation of genes in the operon especially of regulator and promotor

genes alters the mechanism of gene regulation in a definite manner.
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Lecture 11
LAC OPERON AND FINE STRUCTURE OF GENE
The hereditary units which are transmitted from one generation to

the next generation arc called genes. A gene is the fundamental biologic
unit, like the atom which is the fundamental physical unit. Mendel while
explaining the result of his monohybrid and dihybrid crosses, first of all
conceived of the genes as particulate units and referred them by various
names such as hereditary factors or hereditary elements. But his concept
about the gene was entirely hypothetical and he remained ignorant about
the physical and chemical nature of gene.
Even before the rediscovery of Menders laws in 1900, it was
already established that chromosomes have a definite role in the
inheritance because it was found that chromosomes were the only link
between one generation and the next generation and a diploid
chromosome set consists of two morphologically similar sets, one is
derived from the mother and the other from the father at fertilization.
Later on, a parallel behavior among chromosomes and genes was
discovered.
Earlier workers proposed various hypotheses to explain the nature

of genes. For instance, De Vries postulated one gene one character
hypothesis according to which a particular trait of an individual is
controlled by a particular gene. Bateson and Punnett proposed the
presence or absence theory. According to them, in a cross the character
which dominates the other has a determiner, while, the recessive
character has no such determiner. But all the theories were discarded by
Morgan, who produced the particulate gene theory in 1926. He considered
genes as corpuscles, which are arranged in a linear order on the
chromosomes and appear like beads on a string. Each gene was supposed
to be different from ail others. The particulate theory of gene was widely
accepted and supported by cytological observations. But, the discovery of
DNA molecule as a sole carrier of genetic informations base altogether
discarded the Morgan's theory. Therefore, before defining the gene it will
be advisable to consider both the classical as well as modern definitions of
gene.
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Changing Concept of Gene
The concept of gene has been the focal point of study from the
beginning of twentieth century to establish the basis of heredity. The gene
has been examined from two main angles, i.e., (1) genetic view, and (2)
biochemical and molecular view. These aspects are briefly described
below:
(1) A Genetic View
The genetic view or perspective of gene is based mainly on the

Mendelian inheritance, chromosomal theory of inheritance and linkage
studies. Mendel used the term factors for genes and reported that factors
were responsible for transmission of characters from parents to their
offspring. Sutton and Boveri (1903) based on the study of mitosis and
meiosis in higher plants established parallel behaviour of chromosomes
and genes. They reported that both chromosomes and genes segregate
and exhibit random assortment, which clearly demonstrated that genes
are located on chromosomes. The Sutton- Boveri hypothesis is known as
chromosome theory of inheritance.
Morgan based on linkage studies in Drosophila reported that genes

are located on the chromosome in a linear fashion. Some genes do not
assort independently because of linkage between them. He suggested that
recombinants are the result of crossing over. The crossing over increases
if the distance between two genes is more. The number of linkage group
is the same as the number of chromosomes. The chromosome theory and
linkage studies reveal that genes are located on the chromosomes. This
view is sometimes called as bead theory. The important points about the
bead theory are given below:
1. The gene is viewed as a fundamental unit of structure, indivisible

by crossing over. Crossing over occurs between genes but not
within a gene.
2. The gene is considered as a basic unit of change or mutation. It
changes from one allelic form to another, but there are no smaller
components within a gene that can change.
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3. The gene is viewed as a basic unit of function. Parts of a gene, if

they exist, cannot function.
The chromosome has been viewed merely as a vector or transporter of
genes and exists simply to permit their orderly segregation and to shuffle
them in recombination. The bead theory is no more valid for any of the
above three points. Now evidences are available which indicate that: (1) a
gene is divisible (2) part of a gene can mutate, and (3) part of a gene can
function.
The Gene is Divisible

Earlier it was believed that gene is a basic unit of structure which is
indivisible by crossing over. In other words, crossing over occurs between
genes but not within a gene. Now intragenic recombination has been
observed in many organisms which indicates that a gene is divisible. The
intragenic recombination has following two main features.
1. It occurs with rare frequency so that a very large test cross

progeny is required for its detection. Benzer expected to detect
a recombination frequency as low as 10-6, the lowest he actually
found was 10-4 (0.01 x 2 = 0.02%).
2. The alleles in which intragenic recombination occurs are
separated by small distances within a gene and are functionally
related.
Examples of intragenic recombination include bar eye, star
asteroid eye and lozenge eye in Drosophila. The bar locus is briefly
described below. Lozenge eye and star asteroid have been discussed
under pseudoalleles.
Bar Eye in Drosophila
The first case of intragenic recombination was recorded in

Drosophila for bar locus which controls size of eye. The bar locus contains
more than one unit of function. The dominant bar gene in Drosophila
produces slit like eye instead of normal oval eye. Bar phenotype is caused
by tandem duplication of 16A region in X chromosome, which results due
to unequal crossing over. The flies with different dose of 16A region have
different types of eye as follows:
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1. Single 16A region → Wild type oval eye

2. Double 16A region → Bar eye small in size
3. Triple 16A region → Double bar or ultrabar eye very small in size
The homozygous bar eye (B/B) produced both wild and ultra bar types
though at a low frequency which indicated intragenic recombination in
the bar locus but the frequency was much higher than that expected
due to spontaneous mutations.
Part of a Gene Can Function

It was considered earlier that gene is the basic unit of function and
parts of gene, if exist, cannot function. But this concept has been
outdated now. Based on studies on rll locus of T4 phage, Banzer (1955)
concluded that there are three sub divisions of a gene, viz., recon, muton
and cistron. These are briefly described below:
Recon
Recons are the regions (units) within a gene between which
recombinations can occur, but the recombination cannot occur within a
recon. There is a minimum recombination distance within a gene which
separates recons. The map of a gene is completely linear sequence of
recons.
Muton
It is the smallest element within a gene, which can give rise to a

mutant phenotype or mutation. This indicates that part of a gene can
mutate or change. This disproved the bead theory according to which the
entire gene was a mutate or change.
Cistron
It is the largest element within a gene which is the unit of function.

This also nocked down the bead theory according to which entire gene
was the unit of function. The name cistron has been derived from the test
which is performed to know whether two mutants are within the same
cistron on in different cistrons. It is called cis-trans test which is
described below.
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Cis – Trans Test

When two mutations in trans position produce mutant phenotype,
they are in the same cistron. Complementation in trans position
(appearance of wild type) indicates that the mutant sites are in different
cistrons. There is no complementation between mutations within a ciston.
It is now known that some genes consist of only one cistron ; some
consist of two or even more. For example, the mutant miniature (m) and
dusky (dy) both decrease wing size in Drosophila and map in the same
part of X chromosome. But when brought together in dy +/+m
heterozygote, the phenotype is normal which indicates that the locus
concerned with wing size is composed of at least two cistrons.
(2) A Biochemical View
It is now generally believed that a gene is a sequence of

nucleotides in DNA which controls a single polypeptide chain. The different
mutations of a gene may be due to change in single nucleotide at more
than one location in the gene. Crossing over can take place between the
altered nucleotides within a gene. Since the mutant nucleotides are placed
so close together, crossing over is expected within very low frequency.
When several different genes which affect the same trait are present so
close that crossing over is rare between them, the term complex locus is
applied to them. Within the nucleotide sequence of DNA, which
represents a gene, multiple alleles are due to mutations at different points
within the gene.
Fine Structure of Gene
Benzer, in 1955, divided the gene into recon, muton and cistron
which are the units of recombination, mutation and function within a
gene. Several units of this type exist in a gene. In order words, each gene
consists of several units of function, mutation and recombination. The fine
structure of gene deals with mapping of individual gene locus. This is
parallel to the mapping of chromosomes. In chromosome mapping,
various genes are assigned on a chromosome, whereas in case of a gene
several alleles are assigned to the same locus. The individual gene maps
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are prepared with the help of intragenic recombination. Since the

frequency of intragenic recombination is extremely low, very large
population has to be grown to obtain such rare combination. Prokaryotes
are suitable materials for growing large population. In Drosophila, 14
alleles of lozenge gene map at four mutational sites which belong to the
same locus (Green, 1961). Similarly, for rosy eye in Drosophila, different
alleles map at 10 mutational sites of the same locus.
Genes can be classified in various ways. The classification of genes is generally

done on the basis of (1) dominance, (2) interaction, (3) character controlled, (4) effect
on survival, (5) location, (6) movement, (7) nucleotide sequence, (8) sex linkage, (9)
operon model, and (10) role in mutation. A brief classification of genes on the basis
of above criteria is presented below
Classification and brief description of genes

Classification of A brief description
genes
Based on
Dominance
Dominant genes Genes that express in the F 1
Recessive genes Genes whose effect is suppressed in F 1

Based on
Interaction
Epistatic gene A gene that has masking effect on the other gene
controlling the same trait.
Hypostatic gene A gene whose expression is masked by another

gene governing the same trait
Based on Character
Controlled
Major gene A gene that governs qualitative trait. Such genes

have distinct phenotypic effects.
Minor gene A gene which is involved in the expression of

quantitative trait. Effect of such genes cannot be
easily detected.
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Based on Effect on
Survival
Lethal gene A gene which leads to death of its carrier when in

homozygous condition. It may be dominant or
recessive.
Semi lethal gene
A gene that causes mortality of more than 50% of
its carriers.
Sub-vital gene
A gene that causes mortality of less than 50% of
its carriers.
Vital gene
A gene that does not have lethal effect on its
carriers.
Based on Location
Nuclear genes Genes that are found in nuclear genome in the

chromosomes.
Plasma genes Genes that are found in the cytoplasm in

mitochondria and chloroplasts. Also called
cytoplasmic or extranuclear genes.
Based on Position
Normal genes Genes that have a fixed position on the

chromosomes. Most of the genes belong to this
category
Jumping genes
Genes which keep on changing their position on
the chromosome of a genome. Such genes have
been reported in maize.
Based on
Nucleotide
sequence
Normal genes Genes having continuous sequence of nucleotides

which code for a single polypeptide chain.
Split gene A gene having discontinuous sequence of

nucleotides. Such genes have been reported in
some eukaryotes. The intervening sequences do
not code for amino acids.
Pseudo genes
Genes having defective nucleotides which are non-
functional. These genes are defective copies of
some normal genes.
Based on Sex
Linkage
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Sex linked genes Genes which are located on sex or X-

chromosomes.
Sex limited genes Genes which express in one sex only
Sex influenced genes Genes whose expression depends on the sex of

individual e.g., gene for baldness in humans.
Based on Operon
Model
Regulator gene A gene found in lac operon of E.Coli which directs

synthesis of a repressor
Operator gene In lac operon, a gene which control the function of

structural genes.
Promotor gene A gene in lac operon of E.Coli which initiates

mRNA synthesis
Structural genes The genes in lac operon of E.Coli which control the
synthesis of protein through mRNA.
Based on role in
Mutation
Mutable genes Genes which exhibit higher mutation rate than

others e.g., which eye gene is Drosophila.
Mutator genes Genes which enhance the natural mutation rate of

other genes in the same genome e.g., dotted gene
in maize.
Antimutator genes
Genes which decrease the frequency of natural
mutation of other genes in the same genome.
Such genes are found in bacteria and
bacteriophages.
More about Genes
There are some genes which are different from normal genes either
in terms of their nucleotide sequences or functions. Some examples of
such genes are split gene, jumping gene, overlapping gene and pseudo
gene. A brief description of each of these genes is presented below:
Split Genes
Usually a gene has a continuous sequence of nucleotides. In other
words, there is no interruption in the nucleotide sequence of a gene. Such
nucleotide sequence codes for a particular single polypeptide chain.
However, it was observed that the sequence of nucleotides was not
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continuous in case of some genes; the sequences of nucleotides were

interrupted by intervening sequences. Such gene with interrupted
sequence of nucleotides is referred to as split genes or interrupted genes.
Thus, split genes have two types of sequences, viz., normal sequences
and interrupted sequences.
1. Normal Sequence. This represented the sequence of nucleotides

which are included in the mRNA which is translated from DNA of split
gene These sequences code for a particular polypeptide chain and are
known as exons.
2. Interrupted Sequence: The intervening or interrupted sequence of
split gene are known as introns. These sequences do not code for any
peptide chain. Moreover, interrupted sequences are not included into
mRNA which is transcribed from DNA of split genes. The interrupted
sequences are removed from the mRNA during processing of the
same. In other words, the intervening sequences are discarded in
mRNA as they are non-coding sequences. The coding sequences or
exons are joined by ligase enzyme.
The first case of split gene was reported for ovalbumin gene of
chickens. The ovalbumin gene has been reported to consist of seven
intervening sequences.. Later on interrupted sequences (split genes)
were reported for beta globin gene of mice and rabbits, tRNA genes of
yeast and ribosomal genes of Drosophila.
The intervening sequences are determined with the help of R loop

technique. This technique consists of hybridization between mRNA and
DNA of the same gene under ideal conditions, i.e., at high temperature
and high concentration of formamide. The mRNA pairs with single strand
of DNA. The non-coding sequences or intervening sequences of DNA make
loop in such pairing. The number of loops indicates the number of
interrupted sequences and the size of loop indicates length of the
intervening sequence. These loops can be viewed under electron
microscope. The ovalbumin gene has seven interrupted sequences
(introns) and eight coding sequences (exons). The beta globin gene has
been reported to have two intervening sequences, one 550 nucleotides
long and the other 125 nucleotides long.
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The intervening sequences are excised during processing to form

mature mRNA molecule. Thus, about half of the ovalbumin gene is
discarded during processing. Earlier it was believed that there is
colinearity (correspondence) between the nucleotide sequence and the
sequence of amino acids which it specifies. The discovery of split genes
has disproved the concept of colinearity of genes. Now colinearity
between genes and their products is considered as a chance rather than a
rule. Split genes have been reported mostly in eukaryotes.
Jumping Genes
Generally, a gene occupies a specific position on the chromosome
called locus. However in some cases a gene keeps on chaining its position
within the chromosome and also between the chromosomes of the same
genome. Such genes are known as jumping genes or transposons or
transposable elements. The first case of jumping gene was reported by
Barbara Mc-Clintock in maize as early as in 1950. However, her work did
not get recognition for a long time like that of Mendel. Because she was
much ahead of time and this was an unusual finding, people did not
appreciate it for a long time. This concept was recognized in early
seventies and McClintock was awarded Nobel Prize for this work in 1983.
Later on transposable elements were reported in the chromosome

of E. coli and other prokaryotes. In E.coli, some DNA segments were
found moving from one location to other location. Such DNA segments are
detected by their presence at such a position in the nucleotide sequence,
where they were not present earlier. The transposable elements are of
two types, viz, insertion sequence and transposons.
1. Insertion Sequence. There are different types of insertion

sequences each with specific properties. Such sequences do not
specify for protein and are of very short length. Such sequence has
been reported in some bacteria bacteriophages and plasmids.
2. Transposons. These are coding sequences which code for one or
more proteins. They are usually very long sequences of nucleotides
including several thousand base pairs.
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Transposable elements are considered to be associated with

chromosomal changes such as inversion and deletion. They are hot
spots for such changes and are useful tools for the study of
mutagenesis. In eukaryotes, moving DNA segments have been
reported in maize, yeast and Drosophila.
Overlapping Genes
Earlier it was believed that a nucleotide sequence codes only for
one protein. Recent investigations with prokaryotes especially viruses
have proved beyond doubt that some nucleotide sequences (genes) can
code for two or even more proteins. The genes which code for more than
one protein are known as overlapping genes. In case of overlapping
genes, the complete nucleotide sequence codes for one protein and a part
of such nucleotide sequence can code for another protein. Overlapping
genes are found in tumor producing viruses such as φ X 174, SV 40 and
G4, in virus φ X 174 gene A overlaps gene B. In virus SV 40, the same
nucleotide sequence codes for the protein VP 3 and also for the coboxyl –
terminal end of the protein VP2. In virus G4, the gene A overlaps gene B
and gene E overlaps gene D. The gene of this virus also contains some
portions of nucleotide sequences which are common for gene A and gene
C.
Pseudogenes
There are some DNA sequences, especially in eukaryotes, which are
non-functional and defective copies of normal genes. These sequences do
not have any function. Such DNA sequences or genes are known as
pseudogenes. Pseudogenes have been reported in humans, mouse and
Drosophila. The main features of pseudogenes are given below :
1. Pseudogenes are non functional or defective copies of some normal

genes. These genes are found in large numbers.
2. These genes being defective cannot be translated.
3. These genes do not code for protein synthesis, means they do not
have any significance.
4. The well known examples of pseudogenes are alpha and beta globin
pseudogenes of mouse.
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LAC OPERON CONCEPT
History
The term ‘‘operon’’ was first proposed in a short paper in the
Proceedings of the French Academy of Sciences in 1960. From this paper,
the so-called general theory of the operon was developed. This theory
suggested that all genes are controlled by means of operons through a
single feedback regulatory mechanism: repression. Later, it was
discovered that the regulation of genes is a much more complicated
process. Indeed, it is not possible to talk of a general regulatory
mechanism, as there are many, and they vary from operon to operon.
Despite modifications, the development of the operon concept is
considered one of the landmark events in the history of molecular biology.
Components of operon
The structural genes
The structural genes form a single long polycistronic m RNA

molecule and the number of structural genes corresponds to the number
of proteins. Each structural gene is controlled independently and
transcribe mRNA molecule separately, this, depends on substrate to be
utilized. Example: In lac operon three structural genes (Z, Y, A) are
associated with lactose utilization. Beta-galactosidase is the product of lac
Z that cleaves beta (1-4) linkage of lactose & releases the free
monosaccharides. The enzyme permease (a product of lacy) facilitates the
lactose the entry inside the bacterium. The enzyme transacylase is a
product of lac A where no definite role has been assigned. The lac operon
consists of a promoter (p) operator (o) together with structural genes.
The lac operon cannot function in the presence of sugars other than
lactose.
The operator gene
The operator gene is present adjacent to lac Z gene. The operator

gene overlaps the promoter region. The lac repressor protein binds to the
operator invitro & protect part of the promoter region from the digestion
of DNase. The repressor protein binds to the operator & forms an operator
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–repressor complex which in turn physically blocks the transcription of Z,

Y & A genes by preventing the release of RNA polymerase to begin
transcription.
The promoter gene
The promoter gene is long nucleotide &continuous with the operator

gene. The promoter gene lies between the operator &regulator gene, like
operators the promoter region consists of palindromic sequences of
nucleotides (i.e show 2 fold geometry from a point). These palindromic
sequence are recognized by such proteins that have symmetrically
arranged subunits. This section of two fold symmetry is present on the
CRP site(c-AMP receptor protein site that binds to a protein called
CRP).the CRP is encoded by CRP gene, it has been shown experimentally
that CRP gene binds to cAMP (c AMP found in e.coli & other organisms)
molecule & form a cAMP CRP complex. This complex is required for
transcription because it binds to promoter& enhances the attachment of
RNA polymerase to the promoter therefore it increases the transcription
&translation process.
The repressor (regulator) gene
Regulator gene determines the transcription of structural gene. It is

of two types-active & inactive repressor. It codes for amino acids of a
defined repressor protein. After synthesis, the repressor molecules are
diffused from the ribosome & bind to the operator in the absence of an
induces. Finally the path of RNA polymerase is blocked & m RNA is not
transcribed consequently; no protein synthesis occurs .this type of
mechanism occurs in inducible system of active repressor. Moreover when
an inducer is present it binds to repressor proteins 7forms an inducer –
repressor complex. Due to formation of complex the repressor undergoes
changes in the confirmation of shape 7 becomes inactive consequently the
structural genes can synthesize the polycistronic m RNA and later
synthesize enzyme.
In contrast in the reversible system the regulator gene

synthesis repressor protein that is inactive & therefore fails to binds to
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operator, consequently ,proteins are synthesized by the structural genes

.however the repressor protein can be activated in the presence of an co-
repressor. the co-repressor together with repressor proteins forms the
repressor-co repressor complex. This complex binds to operator gene &
blocks the protein synthesis
Types of operon
1. Lactose (Lac) operon
The regulatory mechanism of operon is responsible for the

utilization of lactose as a carbon source that is why it is called as lac
operon. the lactose utilizing system consists of 2 types of components i.e
the structural genes (lacZ, lacy, lacA) the products of which are required
for transport and metabolism of lactose &regulatory genes (lacI, lacP,
lacO).these two components together comprises of lac operon .one of the
most key features is that operon provides a mechanism for the co-
ordinated expression of structural genes controlled by regulatory genes.
Operon shows polarity i.e. the genes Z, Y, A synthesize equally qualities of
3 enzymes beta-galactosidase by lac Z, permease by lac Y & acetylase by
lac A. These are synthesized in an order i.e. beta-galactosidase at first
and acetylase in the last.
Regulation of lac operon
Regulation of the lac operon by repressor is called negative control.

The lac operon is also under positive control by CRP (or cAMP Receptor
Protein; also known as CAP or catabolite activator protein). CRP or CAP is
now thought to be bound to its lac binding site at all times (even during
repression). During induction, the inducer (either the natural inducer,
allolactose, or the synthetic inducer, IPTG, binds to the lac repressor.
Inducer-bound repressor does not bind to operator sites. This allows RNA
polymerase to bind to the promoter and start transcribing the lac operon.
Negative (lac repressor) -------------Bound to DNA-----------Not

bound to DNA
(Type of Control) (Operon
off) (Operon on)
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Positive(CRP protein) --------------- Bound to DNA-----------Not

bound to DNA
(Type of control) (Operon
on) (Operon off)
2. Tryptophan (Trp) operon
The tryptophan operon of E.coli is responsible for the synthesis of

the amino acids tryptophan regulation of this operon occurs in such a way
that when tryptophan is present in the growth medium, Trp operon is not
active but, when adequate trp is present, the transcription of the operon
is inhibited, however when its supply is insufficient transcription occurs,
the Trp is quite different from the lac operon in that trp acts directly in the
repression system rather than as an inducer. Moreover since the trp
operon encodes a set of bio-synthetic caranabolic rather than a catabolic
enzyme neither glu nor c AMP –CAP has a role in the operon activity.
Regulation of Trp Operon
Trp is synthesized in 5 steps each required a particular enzyme.in

E.coli chromosome the genes encoding these enzymes are located
adjacent to one another in the same order as they are used in the bio-
synthetic pathway they are translated from a single polycistronic m RNA
molecule. These genes are called TrpE, TrpP, TrpC, TrpB, TrpA, The TrpE
gene is the first one translated. Adjacent to the Trp E gene are the
promoter, the operator &2 region, called the leader and the attenuated
which are designated as TrpL & TrpA respectively .the repressor gene
TrpR is located quite for from the gene cluster. The regulatory protein of
the repressor system o the TrpR operon is the product of the TrpR gene.
mutations either in this gene or in the operator cause constitute initiation
of transcription of Trp-m RNA on the lac operon. This regulatory protein is
called Trp apo repressor &it does not bind to the operator, unless Trp is
present. The apo repressor &the tryptophan molecule joins together to
form an active trp repressor which binds to the operator. The reaction
scheme is as follows:
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Apo repressor (no trp) ---------------active repressor (transcription

occurs)
Apo repressor+trp------------------active repressor +operator-------

--------- inactive operator (transcription does not occur)
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Lecture 12
ULTRA STRUCTURE OF CELL AND CELL ORGANELLES AND THEIR FUNCTIONS
In 1665, an Englishman by the name of Robert Hooke examined thin slices of cork and
observed that it was composed of numerous little boxes, fitted together like honey comb. Since
these boxes resembled the compartment of monastery he named them as cells. The cork cells
studied by Hooke were really empty boxes; they had lost their living matter, the protoplasm.
After his discovery, the protoplasm in living cells were largely over looked due to its
transparency. Today, with the help of special techniques, we are able to see not only the
protoplasm but also many bodies inside it.
A general outline of a plant cell is as follows:
I. Cell Wall
II. Protoplast
A. Protoplasmic reticulum
1. Cytoplasm
2. Endoplasmic reticulum
3. Ribosomes
4. Mitochondria
5. Golgi apparatus
6. Plastids
7. Nucleus
8. Plant cell vacuoles
9. Peroxisomes
10. Lysosomes
11. spherosomes
B. Non-protoplasmic components
1. Starch grains
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Cell Wall
Plant cells are surrounded by a non living and rigid coat called cell wall. Though the cell
wall is not a living part of the cell, it is an extra cytoplasmic product. Cell walls are significantly
thicker than plasma membranes. It is responsible for the shape of plants and controls the
growth rate of plant cells. It is a structural barrier to some molecules and invading insects. Cell
walls are also a source of energy, fibre and food.
Walls are a layered structure, having three basic portions: intercellular substance or
middle lamella, primary wall and secondary wall. The middle lamella cements together the
primary walls of two contiguous cells and the secondary wall is laid over the primary. The middle
lamella is mainly composed of a pectic compound which mostly appears to be calcium pectate.
The primary wall is largely composed of cellulose and the secondary wall may be of cellulose or
cellulose impregnated with other substances.
Primary cell wall
The main chemical components of the primary plant cell wall include cellulose in the
form of organized microfibrils, a complex carbohydrate made up of several thousand glucose
molecules linked end to end. Cellulose constitutes the bulk of material of cell walls are made. It
is soft, elastic, transparent and readily permeable to water. It is a carbohydrate, being
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composed of three elements – carbon, hydrogen and oxygen. Cotton and linen are nearly pure
cellulose. Cellophane, celluloid, paper rayon, synthetic lacquers and varnish are manufactured
from cellulose. In addition, the cell wall contains two groups of branched polysaccharides, the
pectins and cross-linking glycans. Pectic compounds are exceedingly gelatinous and
viscous. They swell in water. They are found in three forms: insoluble pectose, soluble pectin
and insoluble pectic acid. Organized into a network with the cellulose microfibrils, the cross-
linking glycans increase the tensile strength of the cellulose, whereas the coextensive network
of pectins provides the cell wall with the ability to resist compression. In addition to these
networks, a small amount of protein can be found in all plant primary cell walls. Some of this
protein is thought to increase mechanical strength and part of it consists of enzymes, which
initiate reactions that form, remodel, or breakdown the structural networks of the wall. Such
changes in the cell wall directed by enzymes are particularly important for fruit to ripen and
leaves to fall in autumn.
Gums and mucilages are complex carbohydrates of the cell wall. They can absorb and
retain water, helping in the dispersal of seeds.
One of the most important distinguishing features of plant cells is the presence of a cell
wall. The relative rigidity of the cell wall renders plants sedentary, unlike animals, whose lack of
this type of structure allows their cells more flexibility, which is necessary for locomotion. The
plant cell wall serves a variety of functions. Along with protecting the intracellular contents, the
structure bestows rigidity to the plant, provides a porous medium for the circulation and
distribution of water, minerals, and other nutrients, and houses specialized molecules that
regulate growth and protect the plant from disease.
Secondary cell wall
The secondary plant cell wall, which is often deposited inside the primary cell wall as a
cell matures, sometimes has a composition nearly identical to that of the earlier-developed wall.
More commonly, however, additional substances, especially lignin, are found in the secondary
wall. Lignin is the general name for a group of polymers of aromatic alcohols that are hard and
impart considerable strength to the structure of the secondary wall. Lignin is what provides the
favorable characteristics of wood to the fiber cells of woody tissues and is also common in the
secondary walls of xylem vessels, which are central in providing structural support to plants.
Lignin occurs generally in hard mature tissues, such as those straw and wood. It gives sufficient
rigidity and strength to the plant body. It is permeable to water and solutes. Lignin also makes
plant cell walls less vulnerable to attack by fungi or bacteria, as do cutin, suberin, and other
waxy materials that are sometimes found in plant cell walls.
Cutin is a fatty substance. It forms an external coating, the cuticle on the other cellulose
wall of the epidermal cells of leaves and stems. It is not permeable to water. It is effective in
protecting the foliage from the leaching effects of rain, in reducing water loss from plant surface
and in preventing easy access of the partial parasites. The process of cutin deposition on the
cellulose wall is called cutinisation.
Suberin is a fatty substance which resembles cutin in many qualities. It is permeable to
water and checks the loss of water from the surface of plants. It is an important constituent of
walls of cork cells, but also occur in the internal cells of exodermis and endodermis. The
process of suberin deposition on the cellulose wall is termed suberisation.
Middle lamella
A specialized region associated with the cell walls of plants, and sometimes considered
an additional component of them, is the middle lamella. Rich in pectins, the middle lamella is
shared by neighboring cells and cements them firmly together. Positioned in such a manner,
cells are able to communicate with one another and share their contents through special
conduits. Termed plasmodesmata, these small passages penetrate the middle lamella as well
as the primary and secondary cell walls, providing pathways for transporting cytoplasmic
molecules from one cell to another.
Plasma Membrane
All living cells, prokaryotic and eukaryotic, have a plasma membrane that encloses their
contents and serves as a semi-porous barrier to the outside environment. The membrane acts
as a boundary, holding the cell constituents together and keeping other substances from
entering. The plasma membrane is permeable to specific molecules, however, and allows
nutrients and other essential elements to enter the cell and waste materials to leave the cell.
Small molecules, such as oxygen, carbon dioxide, and water, are able to pass freely across the
membrane, but the passage of larger molecules, such as amino acids and sugars, is carefully
regulated.
According to the accepted current theory, known as the fluid mosaic model, the plasma
membrane is composed of a double layer (bilayer) of lipids, oily substances found in all cells.
Most of the lipids in the bilayer can be more precisely described as phospholipids, that is,
lipids that feature a phosphate group at one end of each molecule. Phospholipids are
characteristically hydrophilic ("water-loving") at their phosphate ends and hydrophobic
("water-fearing") along their lipid tail regions. In each layer of a plasma membrane, the
hydrophobic lipid tails are oriented inwards and the hydrophilic phosphate groups are aligned so
they face outwards, either toward the aqueous cytosol of the cell or the outside environment.
Phospholipids tend to spontaneously aggregate by this mechanism whenever they are exposed
to water.
Within the phospholipid bilayer of the plasma membrane, many diverse proteins are
embedded, while other proteins simply adhere to the surfaces of the bilayer. Some of these
proteins, primarily those that are at least partially exposed on the external side of the
membrane, have carbohydrates attached to their outer surfaces and are, therefore, referred to
as glycoproteins. The positioning of proteins along the plasma membrane is related in part to
the organization of the filaments that comprise the cytoskeleton, which help anchor them in
place. The arrangement of proteins also involves the hydrophobic and hydrophilic regions found
on the surfaces of the proteins: hydrophobic regions associate with the hydrophobic interior of
the plasma membrane and hydrophilic regions extend past the surface of the membrane into
either the inside of the cell or the outer environment.
Plasma membrane proteins function in several different ways. Many of the proteins play
a role in the selective transport of certain substances across the phospholipid bilayer, either
acting as channels or active transport molecules. Others function as receptors, which bind
information-providing molecules, such as hormones, and transmit corresponding signals based
on the obtained information to the interior of the cell. Membrane proteins may also exhibit
enzymatic activity, catalyzing various reactions related to the plasma membrane.
Plasmodesmata
Plasmodesmata (singular, plasmodesma) are small channels that directly connect the
cytoplasm of neighboring plant cells to each other, establishing living bridges between cells. The
plasmodesmata, which penetrate both the primary and secondary cell walls allow certain
molecules to pass directly from one cell to another and are important in cellular communication.
Due to the presence of plasmodesmata, plant cells can be considered to form a

synctium, or multinucleate mass with cytoplasmic continuity. Somewhat cylindrical in shape,
plasmodesmata are lined with the plasma membrane so all connected cells are united through
essentially one continuous cell membrane. A majority of plasmodesmata also contain a narrow
tube-like structure called the desmotubule, which is derived from the smooth endoplasmic
reticulum of the connected cells. The desmotubule does not completely fill the plasmodesma
and, consequently, a ring of shared cytoplasm is located between it and the inner surface of the
membrane-lined channel. Plasmodesmata typically form during cell division when parts of the
endoplasmic reticulum of the parent cell get trapped in the new cell wall that is produced to
create daughter cells. Thousands of plasmodesmata may be formed that connect the daughter
cells to one another.
It is widely thought that by constricting and dilating the openings at the ends of the
plasmodesmata, plants cells regulate the passage of small molecules, such as sugars, salts,
and amino acids, though this mechanism is not yet well understood. Yet, it is known that in
some cases the size restrictions on molecule passage between cells can be overcome. By
binding to parts of the plasmodesmata, special proteins and some viruses are able to increase
the diameter of the channels enough for unusually large molecules to pass through.
Cytoplasm
Part of plant cells outside the nucleus (and outside the large vacuole of plant cells) is
called cytoplasm. Strictly speaking, this includes all the organelles (mitochondria, chloroplasts,
and so on) and is the area in which most cell activities take place. However, cytoplasm is often
used to refer to the jellylike matter in which the organelles are embedded (correctly termed the
cytosol). Most of the activities in the cytoplasm are chemical reactions (metabolism), for
example, protein synthesis.
In many cells, the cytoplasm is made up of two parts: the ectoplasm (or plasmagel), a dense
gelatinous outer layer concerned with cell movement, and the endoplasm (or plasmasol), a
more fluid inner part where most of the organelles are found. The semifluid medium between
the nucleus and the plasma membrane is called cytosol.
The Endoplasmic Reticulum
The endoplasmic reticulum (ER) is a network of flattened sacs and branching tubules
that extends throughout the cytoplasm in plant and animal cells. These sacs and tubules are all
interconnected by a single continuous membrane so that the organelle has only one large,
highly convoluted and complexly arranged lumen (internal space). Usually referred to as the
endoplasmic reticulum cisternal space, the lumen of the organelle often takes up more than 10
percent of the total volume of a cell. The endoplasmic reticulum membrane allows molecules to
be selectively transferred between the lumen and the cytoplasm, and since it is connected to the
double-layered nuclear envelope, it further provides a pipeline between the nucleus and the
cytoplasm.
The endoplasmic reticulum manufactures, processes, and transports a wide variety of

biochemical compounds for use inside and outside of the cell. Consequently, many of the
proteins found in the cisternal space of the endoplasmic reticulum lumen are there only
transiently as they pass on their way to other locations. Other proteins, however, are targeted to
constantly remain in the lumen and are known as endoplasmic reticulum resident proteins.
These special proteins, which are necessary for the endoplasmic reticulum to carry out its
normal functions, contain a specialized retention signal consisting of a specific sequence of
amino acids that enables them to be retained by the organelle. An example of an important
endoplasmic reticulum resident protein is the chaperone protein known as BiP (formally: the
chaperone immunoglobulin-binding protein), which identifies other proteins that have been
improperly built or processed and keeps them from being sent to their final destinations.
There are two basic kinds of endoplasmic reticulum morphologies: rough and smooth.
The surface of rough endoplasmic reticulum is covered with ribosomes, giving it a bumpy
appearance when viewed through the microscope. This type of endoplasmic reticulum is
involved mainly with the production and processing of proteins that will be exported, or secreted,
from the cell.
The smooth endoplasmic reticulum in most cells is much less extensive than the rough
endoplasmic reticulum and is sometimes alternatively termed transitional. Smooth
endoplasmic reticulum is chiefly involved, however, with the production of lipids (fats), building
blocks for carbohydrate metabolism, and the detoxification of drugs and poisons. Therefore, in
some specialized cells, such as those that are occupied chiefly in lipid and carbohydrate
metabolism (brain and muscle) or detoxification (liver), the smooth endoplasmic reticulum is
much more extensive and is crucial to cellular function. Smooth endoplasmic reticulum also
plays a role in various cellular activities through its storage of calcium and involvement in
calcium metabolism.
Ribosomes
All living cells contain ribosomes, tiny organelles composed of approximately 60 percent
ribosomal RNA (rRNA) and 40 percent protein. However, though they are generally described
as organelles, it is important to note that ribosomes are not bound by a membrane and are
much smaller than other organelles.
Ribosomes are mainly found bound to the endoplasmic reticulum and the nuclear
envelope, as well as freely scattered throughout the cytoplasm, depending upon whether the
cell is plant, animal, or bacteria.
In 2000, the complete three-dimensional structure of the large and small subunits of a
ribosome was established. Evidence based on this structure suggests, as had long been
assumed, that it is the rRNA that provides the ribosome with its basic formation and
functionality, not proteins. Apparently the proteins in a ribosome help fill in structural gaps and
enhance protein synthesis, although the process can take place in their absence, albeit at a
much slower rate.
The units of a ribosome are often described by their Svedberg (s) values, which are
based upon their rate of sedimentation in a centrifuge. The ribosomes in a eukaryotic cell
generally have a Svedberg value of 80S and are comprised of 40s and 60s subunits.
Prokaryotic cells, on the other hand, contain 70S ribosomes, each of which consists of a 30s
and a 50s subunit. As demonstrated by these values, Svedberg units are not additive, so the
values of the two subunits of a ribosome do not add up to the Svedberg value of the entire
organelle. This is because the rate of sedimentation of a molecule depends upon its size and
shape, rather than simply its molecular weight.
Protein synthesis requires the assistance of two other kinds of RNA molecules in
addition to rRNA. Messenger RNA (mRNA) provides the template of instructions from the
cellular DNA for building a specific protein. Transfer RNA (tRNA) brings the protein building
blocks, amino acids, to the ribosome. There are three adjacent tRNA binding sites on a
ribosome: the aminoacyl binding site for a tRNA molecule attached to the next amino acid in
the protein, the peptidyl binding site for the central tRNA molecule containing the growing
peptide chain, and an exit binding site to discharge used tRNA molecules from the ribosome.
Once the protein backbone amino acids are polymerized, the ribosome releases the protein and
it is transported to the cytoplasm in prokaryotes or to the Golgi apparatus in eukaryotes. There,
the proteins are completed and released inside or outside the cell. Ribosomes are very efficient
organelles. A single ribosome in a eukaryotic cell can add 2 amino acids to a protein chain
every second. In prokaryotes, ribosomes can work even faster, adding about 20 amino acids to
a polypeptide every second.
In addition to the most familiar cellular locations of ribosomes, the organelles can also be found
inside mitochondria and the chloroplasts of plants. The similarity of mitochondrial and
chloroplast ribosomes to prokaryotic ribosomes is generally considered strong supportive
evidence that mitochondria and chloroplasts evolved from ancestral prokaryotes.
Mitochondria
Mitochondria are rod-shaped organelles that can be considered the power generators of
the cell, converting oxygen and nutrients into adenosine triphosphate (ATP). ATP is the
chemical energy "currency" of the cell that powers the cell's metabolic activities.
The number of mitochondria present in a cell depends upon the metabolic requirements
of that cell, and may range from a single large mitochondrion to thousands of the organelles.
Mitochondria, which are found in nearly all eukaryotes, including plants, animals, fungi, and
protists, are large enough to be observed with a light microscope and were first discovered in
the 1800s. The name of the organelles was coined to reflect the way they looked to the first
scientists to observe them, stemming from the Greek words for "thread" and "granule." For
many years after their discovery, mitochondria were commonly believed to transmit hereditary
information. It was not until the mid-1950s when a method for isolating the organelles intact was
developed that the modern understanding of mitochondrial function was worked out.
The elaborate structure of a mitochondrion is very important to the functioning of the
organelle. Two specialized membranes encircle each mitochondrion present in a cell, dividing
the organelle into a narrow intermembrane space and a much larger internal matrix, each of
which contains highly specialized proteins. The outer membrane of a mitochondrion contains
many channels formed by the protein porin and acts like a sieve, filtering out molecules that are
too big. Similarly, the inner membrane, which is highly convoluted so that a large number of
infoldings called cristae are formed, also allows only certain molecules to pass through it and is
much more selective than the outer membrane. To make certain that only those materials
essential to the matrix are allowed into it, the inner membrane utilizes a group of transport
proteins that will only transport the correct molecules. Together, the various compartments of a
mitochondrion are able to work in harmony to generate ATP in a complex multi-step process.
Mitochondria are generally oblong organelles, which range in size between 1 and 10
micrometers in length, and occur in numbers that directly correlate with the cell's level of
metabolic activity. The organelles are quite flexible, however, studies have demonstrated that
mitochondria change shape rapidly and move about in the cell almost constantly. Movements of
the organelles appear to be linked in some way to the microtubules present in the cell, and are
probably transported along the network with motor proteins. Consequently, mitochondria may
be organized into lengthy traveling chains, packed tightly into relatively stable groups, or appear
in many other formations based upon the particular needs of the cell and the characteristics of
its microtubular network.
Golgi apparatus
The Golgi apparatus (GA), also called Golgi body or Golgi complex and found
universally in both plant and animal cells, is typically comprised of a series of five to eight cup-
shaped, membrane-covered sacs called cisternae that look something like a stack of deflated
balloons. In some unicellular flagellates, however, as many as 60 cisternae may combine to
make up the Golgi apparatus. Similarly, the number of Golgi bodies in a cell varies according to
its function. Animal cells generally contain between ten and twenty Golgi stacks per cell, which
are linked into a single complex by tubular connections between cisternae. This complex is
usually located close to the cell nucleus.
Due to its relatively large size, the Golgi apparatus was one of the first organelles ever
observed. In 1897, an Italian physician named Camillo Golgi, who was investigating the nervous
system by using a new staining technique he developed (and which is still sometimes used
today; known as Golgi staining or Golgi impregnation), observed in a sample under his light
microscope a cellular structure that he termed the internal reticular apparatus. Soon after he
publicly announced his discovery in 1898, the structure was named after him, becoming
universally known as the Golgi apparatus. Yet, many scientists did not believe that what Golgi
observed was a real organelle present in the cell and instead argued that the apparent body
was a visual distortion caused by staining. The invention of the electron microscope in the
twentieth century finally confirmed that the Golgi apparatus is a cellular organelle.
The Golgi apparatus is often considered the distribution and shipping department for the
cell's chemical products. It modifies proteins and lipids (fats) that have been built in the
endoplasmic reticulum and prepares them for export outside of the cell or for transport to other
locations in the cell. Proteins and lipids built in the smooth and rough endoplasmic reticulum bud
off in tiny bubble-like vesicles that move through the cytoplasm until they reach the Golgi
complex. The vesicles fuse with the Golgi membranes and release their internally stored
molecules into the organelle. Once inside, the compounds are further processed by the Golgi
apparatus, which adds molecules or chops tiny pieces off the ends. When completed, the
product is extruded from the GA in a vesicle and directed to its final destination inside or outside
the cell. The exported products are secretions of proteins or glycoproteins that are part of the
cell's function in the organism. Other products are returned to the endoplasmic reticulum or may
undergo maturation to become lysosomes.
The modifications to molecules that take place in the Golgi apparatus occur in an orderly
fashion. Each Golgi stack has two distinct ends, or faces. The cis face of a Golgi stack is the
end of the organelle where substances enter from the endoplasmic reticulum for processing,
while the trans face is where they exit in the form of smaller detached vesicles. Consequently,
the cis face is found near the endoplasmic reticulum, from whence most of the material it
receives comes, and the trans face is positioned near the plasma membrane of the cell, to
where many of the substances it modifies are shipped. The chemical make-up of each face is
different and the enzymes contained in the lumens (inner open spaces) of the cisternae
between the faces are distinctive.
Proteins, carbohydrates, phospholipids, and other molecules formed in the endoplasmic
reticulum are transported to the Golgi apparatus to be biochemically modified during their
transition from the cis to the trans poles of the complex. Enzymes present in the Golgi lumen
modify the carbohydrate (or sugar) portion of glycoproteins by adding or subtracting individual
sugar monomers. In addition, the Golgi apparatus manufactures a variety of macromolecules on
its own, including a variety of polysaccharides. The Golgi complex in plant cells produces
pectins and other polysaccharides specifically needed by for plant structure and metabolism.
The products exported by the Golgi apparatus through the trans face eventually fuse with the
plasma membrane of the cell. Among the most important duties of the Golgi apparatus is to sort
the wide variety of macromolecules produced by the cell and target them for distribution to their
proper location. Specialized molecular identification labels or tags, such as phosphate groups,
are added by the Golgi enzymes to aid in this sorting effort.
Plastids
In most plant cells structures called plastids are found. They are found in the cytoplasmic
matrix of plant cells only. These structures are generally spherical or ovoid in shape and they
are clearly visible in living cells. 3 types of plastids found in plant cells:
Chromoplasts
Chromoplasts are red, yellow or orange in colour and are found in petals of flowers and
in fruit. Their colour is due to two pigments, carotene and xanthophyll.
Functions
the primary function in the cells of flowers is to attract agents of pollination,
and in fruit to attract agents of dispersal.
Leucoplasts
Leucoplasts are colourless plastids and occur in plant cells not exposed to light, such as roots
and seeds. They are colourless due the absent of pigments.
Functions
leucoplasts are the centers of starch grain formation;
they are also involved in the synthesis of oils and proteins.
Chloroplasts
Chloroplasts are probably the most important among the plastids since they are directly
involved in photosynthesis. They are usually situated near the surface of the cell and occur in
those parts that receive sufficient light, e.g. the palisade cells of leaves. The green colour of
chloroplasts is caused by the green pigment chlorophyll.
One of the most widely recognized and important characteristics of plants is their ability
to conduct photosynthesis, in effect, to make their own food by converting light energy into
chemical energy. This process occurs in almost all plant species and is carried out in
specialized organelles known as chloroplasts. All of the green structures in plants, including
stems and unripened fruit, contain chloroplasts, but the majority of photosynthesis activity in
most plants occurs in the leaves. On the average, the chloroplast density on the surface of a
leaf is about one-half million per square millimeter.
All plastids develop from tiny organelles found in the immature cells of plant meristems
(undifferentiated plant tissue) termed proplastids, and those of a particular plant species
contain copies of the same circular genome. The disparities between the various types of
plastids are based upon the needs of the differentiated cells they are contained in, which may
be influenced by environmental conditions, such as whether light or darkness surrounds a leaf
as it grows.
The ellipsoid-shaped chloroplast is enclosed in a double membrane and the area
between the two layers that make up the membrane is called the intermembrane space. The
outer layer of the double membrane is much more permeable than the inner layer, which
features a number of embedded membrane transport proteins. Enclosed by the chloroplast
membrane is the stroma, a semi-fluid material that contains dissolved enzymes and comprises
most of the chloroplast's volume. Since, like mitochondria, chloroplasts possess their own
genomes (DNA), the stroma contains chloroplast DNA and special ribosomes and RNAs as
well. In higher plants, lamellae, internal membranes with stacks (each termed a granum) of
closed hollow disks called thylakoids, are also usually dispersed throughout the stroma. The
numerous thylakoids in each stack are thought to be connected via their lumens (internal
spaces).
Light travels as packets of energy called photons and is absorbed in this form by light-
absorbing chlorophyll molecules embedded in the thylakoid disks. When these chlorophyll
molecules absorb the photons, they emit electrons, which they obtain from water (a process that
results in the release of oxygen as a byproduct). The movement of the electrons causes
hydrogen ions to be propelled across the membrane surrounding the thylakoid stack, which
consequently initiates the formation of an electrochemical gradient that drives the stroma's
production of adenosine triphosphate (ATP). ATP is the chemical energy "currency" of the cell
that powers the cell's metabolic activities. In the stroma, the light-independent reactions of
photosynthesis, which involve carbon fixation, occur, and low-energy carbon dioxide is
transformed into a high-energy compound like glucose.
NUCLEUS
The nucleus is a highly specialized organelle that serves as the information processing
and administrative center of the cell. This organelle has two major functions: it stores the cell's
hereditary material, or DNA, and it coordinates the cell's activities, which include growth,
intermediary metabolism, protein synthesis, and reproduction (cell division).
The spherical nucleus typically occupies about 10 percent of a eukaryotic cell's volume,
making it one of the cell's most prominent features. A double-layered membrane, the nuclear
envelope, separates the contents of the nucleus from the cellular cytoplasm. The envelope is
riddled with holes called nuclear pores that allow specific types and sizes of molecules to pass
back and forth between the nucleus and the cytoplasm. It is also attached to a network of
tubules and sacs, called the endoplasmic reticulum, where protein synthesis occurs, and is
usually studded with ribosome.
The semifluid matrix found inside the nucleus is called nucleoplasm. Within the
nucleoplasm, most of the nuclear material consists of chromatin, the less condensed form of the
cell's DNA that organizes to form chromosomes during mitosis or cell division. The nucleus also
contains one or more nucleoli, organelles that synthesize protein-producing macromolecular
assemblies called ribosomes, and a variety of other smaller components, such as Cajal bodies,
GEMS (Gemini of coiled bodies), and interchromatin granule clusters.
Chromatin and Chromosomes
Packed inside the nucleus of every human cell is nearly 6 feet of DNA, which is divided
into 46 individual molecules, one for each chromosome and each about 1.5 inches long. For
DNA to function, it is combined with proteins and organized into a precise, compact structure, a
dense string-like fiber called chromatin.
The Nucleolus
The nucleolus is a membrane-less organelle within the nucleus that manufactures
ribosomes, the cell's protein-producing structures. The nucleolus looks like a large dark spot
within the nucleus. A nucleus may contain up to four nucleoli, but within each species the
number of nucleoli is fixed. After a cell divides, a nucleolus is formed when chromosomes are
brought together into nucleolar organizing regions. During cell division, the nucleolus
disappears.
The Nuclear Envelope
The nuclear envelope is a double-layered membrane that encloses the contents of the
nucleus during most of the cell's lifecycle. The space between the layers is called the
perinuclear space and appears to connect with the rough endoplasmic reticulum. The envelope
is perforated with tiny holes called nuclear pores. These pores regulate the passage of
molecules between the nucleus and cytoplasm, permitting some to pass through the membrane,
but not others. The inner surface has a protein lining called the nuclear lamina, which binds to
chromatin and other nuclear components. During mitosis, or cell division, the nuclear envelope
disintegrates, but reforms as the two cells complete their formation and the chromatin begins to
unravel and disperse.
Nuclear Pores
The nuclear envelope is perforated with holes called nuclear pores. These pores
regulate the passage of molecules between the nucleus and cytoplasm, permitting some to
pass through the membrane, but not others. Building blocks for building DNA and RNA are
allowed into the nucleus as well as molecules that provide the energy for constructing genetic
material.
Peroxisomes
Microbodies are a diverse group of organelles that are found in the cytoplasm of almost
all cells, roughly spherical, and bound by a single membrane. There are several types of
microbodies, including lysosomes, but peroxisomes are the most common. All eukaryotes are
comprised of one or more cells that contain peroxisomes. The organelles were first discovered
by the Belgian scientist Christian de Duve, who also discovered lysosomes.
Peroxisomes contain a variety of enzymes, which primarily function together to rid the
cell of toxic substances, and in particular, hydrogen peroxide (a common byproduct of cellular
metabolism). These organelles contain enzymes that convert the hydrogen peroxide to water,
rendering the potentially toxic substance safe for release back into the cell. Some types of
peroxisomes, such as those in liver cells, detoxify alcohol and other harmful compounds by
transferring hydrogen from the poisons to molecules of oxygen (a process termed oxidation).
Others are more important for their ability to initiate the production of phospholipids, which are
typically used in the formation of membranes.
Plant Cell Vacuoles

Vacuoles are membrane-bound sacs within the cytoplasm of a cell that function in
several different ways. In mature plant cells, vacuoles tend to be very large and are extremely
important in providing structural support, as well as serving functions such as storage, waste
disposal, protection, and growth. Many plant cells have a large, single central vacuole that
typically takes up most of the room in the cell (80 percent or more). Vacuoles in animal cells,
however, tend to be much smaller, and are more commonly used to temporarily store materials
or to transport substances.
The central vacuole in plant cells is enclosed by a membrane termed the tonoplast, an
important and highly integrated component of the plant internal membrane network
(endomembrane) system. This large vacuole slowly develops as the cell matures by fusion of
smaller vacuoles derived from the endoplasmic reticulum and Golgi apparatus. Because the
central vacuole is highly selective in transporting materials through its membrane, the chemical
palette of the vacuole solution (termed the cell sap) differs markedly from that of the
surrounding cytoplasm. For instance, some vacuoles contain pigments that give certain flowers
their characteristic colors. The central vacuole also contains plant wastes that taste bitter to
insects and animals, while developing seed cells use the central vacuole as a repository for
protein storage.
Among its roles in plant cell function, the central vacuole stores salts, minerals, nutrients,
proteins, pigments, helps in plant growth, and plays an important structural role for the plant.
Under optimal conditions, the vacuoles are filled with water to the point that they exert a
significant pressure against the cell wall. This helps maintain the structural integrity of the plant,
along with the support from the cell wall, and enables the plant cell to grow much larger without
having to synthesize new cytoplasm. In most cases, the plant cytoplasm is confined to a thin
layer positioned between the plasma membrane and the tonoplast, yielding a large ratio of
membrane surface to cytoplasm.
The structural importance of the plant vacuole is related to its ability to control turgor
pressure. Turgor pressure dictates the rigidity of the cell and is associated with the difference
between the osmotic pressure inside and outside of the cell. Osmotic pressure is the pressure
required to prevent fluid diffusing through a semipermeable membrane separating two solutions
containing different concentrations of solute molecules. The response of plant cells to water is a
prime example of the significance of turgor pressure. When a plant receives adequate amounts
of water, the central vacuoles of its cells swell as the liquid collects within them, creating a high
level of turgor pressure, which helps maintain the structural integrity of the plant, along with the
support from the cell wall. In the absence of enough water, however, central vacuoles shrink
and turgor pressure is reduced, compromising the plant's rigidity so that wilting takes place.
Plant vacuoles are also important for their role in molecular degradation and storage.
Sometimes these functions are carried out by different vacuoles in the same cell, one serving as
a compartment for breaking down materials (similar to the lysosomes found in animal cells), and
another storing nutrients, waste products, or other substances. Several of the materials
commonly stored in plant vacuoles have been found to be useful for humans, such as opium,
rubber, and garlic flavoring, and are frequently harvested. Vacuoles also often store the
pigments that give certain flowers their colors, which aid them in the attraction of bees and other
pollinators, but also can release molecules that are poisonous, odoriferous, or unpalatable to
various insects and animals, thus discouraging them from consuming the plant.
Amyloplast (Starch Grain)
A membrane-bound organelle containing concentric layers of starch (amylopectin). This
organelle is commonly found in subterranean storage organs, such as tubers (potatoes), corms
(taro & dasheen), and storage roots (sweet potatoes). Amyloplasts are also found in bananas
and other fruits.
Lecture 13
Study of chromosome structure, morphology, number and types -

Karyotype and Idiogram
A chromosome is a structure that occurs within cells and that contains the cell's genetic
material. That genetic material, which determines how an organism develops, is a molecule of
deoxyribonucleic acid (DNA). A molecule of DNA is a very long, coiled structure that contains
many identifiable subunits known as genes. In prokaryotes, or cells without a nucleus, the
chromosome is merely a circle of DNA. In eukaryotes, or cells with a distinct nucleus,
chromosomes are much more complex in structure.
Historical background
The terms chromosome and gene were used long before biologists really understood
what these structures were. When the Austrian monk and biologist Gregor Mendel (1822–1884)
developed the basic ideas of heredity, he assumed that genetic traits were somehow
transmitted from parents to offspring in some kind of tiny "package." That package was later
given the name "gene." When the term was first suggested, no one had any idea as to what a
gene might look like. The term was used simply to convey the idea that traits are transmitted
from one generation to the next in certain discrete units.
The term "chromosome" was first suggested in 1888 by the German anatomist Heinrich
Wilhelm Gottfried von Waldeyer-Hartz (1836–1921). Waldeyer-Hartz used the term to describe
certain structures that form during the process of cell division (reproduction).
One of the greatest breakthroughs in the history of biology occurred in 1953 when
American biologist James Watson and English chemist Francis Crick discovered the chemical
structure of a class of compounds known as deoxyribonucleic acids (DNA). The Watson and
Crick discovery made it possible to express biological concepts (such as the gene) and
structures (such as the chromosome) in concrete chemical terms.
According to the classical cytological studies, each chromosome structurally consists of a
limiting membrane called pellicle, an amorphous matrix and two very thin, highly coiled filaments
called chromonema or chromonemata. Each chromonemata is 800A 0 thick and contains 8-
microfibriis, each of which in its turn contains two double helics of DNA. Both chromonematae
remain intimately coiled in spiral manner with each other and have a series of microscopically
visible bead-like swelling along its length called chromomeres. The early geneticists have
attached great significance to the chromomeres and errorneously considered them as
hereditary unit, hereditary or Mendelian factors or genes; but modern cytological investigations
have confirmed that the chromomeres are not genes but the regions of super-imposed coils.
The recent cytological findings have also condemned the view that chromosomes have pellicle,
matrix and chromonemata.
A. Nucleolus organizer B. Chromosome C. Nucleolus
The structure of chromosomes and genes

A chromosome is an organized structure of DNA and protein that is found in cells. A
chromosome is a single piece of coiled DNA containing many genes, regulatory elements and
other nucleotide sequences. Chromosomes also contain DNA-bound proteins, which serve to
package the DNA and control its functions. The word chromosome comes from the Greek
chroma - color and soma - body due to their property of being very strongly stained by particular
dyes. Chromosomes vary widely between different organisms. The DNA molecule may be
circular or linear, and can be composed of 10,000 to 1,000,000,000 nucleotides in a long chain.
Typically eukaryotic cells (cells with nuclei) have large linear chromosomes and prokaryotic cells
(cells without defined nuclei) have smaller circular chromosomes, although there are many
exceptions to this rule.
Today we know that a chromosome contains a single molecule of DNA along with
several kinds of proteins. A molecule of DNA, in turn, consists of thousands and thousands of
subunits, known as nucleotides, joined to each other in very long chains. A single molecule of
DNA within a chromosome may be as long as 8.5 centimeters (3.3 inches). To fit within a
chromosome, the DNA molecule has to be twisted and folded into a very complex shape.
Each chromosome has a constriction point called the centromere, which divides the
chromosome into two sections, or “arms.” The short arm of the chromosome is labeled the “p
arm.” The long arm of the chromosome is labeled the “q arm.” The location of the centromere on
each chromosome gives the chromosome its characteristic shape, and can be used to help
describe the location of specific genes.
The arrangement of packets of genetic information in a chromosome is as follows:
Furthermore, cells may contain more than one type of chromosome; for example, mitochondria
in most eukaryotes and chloroplasts in plants have their own small chromosomes. The following
are the different types of chromosomes
Viral Chromosomes
The chromosomes of viruses are called viral chromosomes. They occur singly in a viral
species and chemically may contain either DNA or RNA. The DNA containing viral
chromosomes may be either of linear shape (e.g., T2, T3, T4, T5, bacteriophages) or circular
shape (e.g., most animal viruses and certain bacteriophages). The RNA containing viral
chromosomes are composed of a linear, single-stranded RNA molecule and occur in some
animal viruses (e.g., poliomyelitis virus, influenza virus, etc.); most plant viruses, (e.g., tobacco
mosaic virus, TMV) and some bacteriophages. Both types of viral chromosomes are either
tightly packed within the capsids of mature virus particles (virons) or occur freely inside the host
cell.
Prokaryotic Chromosomes
The prokaryotes usually consists of a single giant and circular chromosome in each of
their nucloids. Each prokaryotic chromosome consists of a single circular, double-stranded DNA
molecule; but has no protein and RNA around the DNA molecule like eukaryotes. Different
prokaryotic species have different sizes of chromosome.
Eukaryotic Chromosomes
The eukaryotic chromosomes differ from the prokaryotic chromosomes in morphology,
chemical composition and molecular structure. The eukaryotes (plants and animals) usually
contain much more genetic informations than the viruses and prokaryotes, therefore, contain a
great amount of genetic material, DNA molecule which here may not occur as a single unit, but,
as many units called chromosomes. Different species of eukaryotes have different but always
constant and characteristic number of chromosomes. In eukaryotes, nuclear chromosomes are
packaged by proteins into a condensed structure called chromatin. This allows the very long
DNA molecules to fit into the cell nucleus. The shape of the eukaryotic chromosomes is
changeable from phase to phase in the continuous process of the cell growth and cell division.
Chromosomes are the essential unit for cellular division and must be replicated, divided, and
passed successfully to their daughter cells so as to ensure the genetic diversity and survival of
their progeny. They are thin, coiled, elastic, contractile thread-like structures during the
interphase (when no division of cell occurs) and are called chromatin threads which under low
magnification look like a compact stainable mass, often called as chromatin substance or
material. During metaphase stage of mitosis and prophase of meiosis, these chromatin threads
become highly coiled and folded to form compact and individually distinct ribbon-shaped
chromosomes. These chromosomes contain a clear zone called kinetochore or centromere
along their length.
Eukaryotes (cells with nuclei such as plants, yeast, and animals) possess multiple large
linear chromosomes contained in the cell's nucleus. Each chromosome has one centromere,
with one or two arms projecting from the centromere, although, under most circumstances,
these arms are not visible as such. In addition, most eukaryotes have a small circular
mitochondrial genome, and some eukaryotes may have additional small circular or linear
cytoplasmic chromosomes.
The number and position of centromeres is variable, but is definite in a specific
chromosome of all the cells and in all the individuals of the same species. Thus, according to
the number of the centromere the eukaryotic chromosomes may be acentric (without any
centromere), mono centric (with one centromere), dicentric (with two centromeres) or
polycentric (with more than two centromeres). The centromere has small granules or spherules
and divides the chromosomes into two or more equal or unequal chromosomal arms.
According to the position of the centromere, the eukaryotic chromosomes may be rod-
shaped (telocentric and acrocentric), J-shaped (submetacentric) and V-shaped (metacentric)
During the cell divisions the microtubules of the spindle are get attached with the chromosomal
centromeres and move them towards the opposite poles of cell. Beside centromere, the
chromosomes may bear terminal unipolar segments called telomeres. Certain chromosomes
contain an
additional
specialized segment, the nucleolus organizer, which is associated with the nucleolus.
Position of the centromere in (A) metacentric; (B) submetacentric; (C) acrocentric; and
(D) telocentric chromosomes.
A. Centromere
Acrocentric Telocentric Metacentric Sub metacentric
In the nuclear chromosomes of eukaryotes, the uncondensed DNA exists in a semi-
ordered structure, where it is wrapped around histones (structural proteins), forming a
composite material called chromatin. Chromatin is the complex of DNA and protein found in the
eukaryotic nucleus which packages chromosomes. The structure of chromatin varies
significantly between different stages of the cell cycle, according to the requirements of the
DNA.
Interphase chromatin
During interphase (the period of the cell cycle where the cell is not dividing), two types of
chromatin can be distinguished. The density of the chromatin that makes up each chromosome
(that is, how tightly it is packed) varies along the length of the chromosome.
dense regions are called heterochromatin
less dense regions are called euchromatin.
• Euchromatin, which consists of DNA that is active, e.g., being expressed as protein.
• Heterochromatin, which consists of mostly inactive DNA. It seems to serve structural
purposes during the chromosomal stages. Heterochromatin can be further distinguished
into two types:
o Constitutive heterochromatin, which is never expressed. It is located around the
centromere and usually contains repetitive sequences.
o Facultative heterochromatin, which is sometimes expressed.

Individual chromosomes cannot be distinguished at this stage - they appear in the nucleus as a
homogeneous tangled mix of DNA and protein.
Diploids and Haploids
In contrast to prokaryotes, most eukaryote are diploids, i.e., each somatic cell of them
contains one set of chromosomes inherited from the maternal (female) parent and a comparable
set of chromosomes (called homologous chromosomes) from the paternal (male) parent. The
number of chromosomes in a dual set of a diploid somatic cell is called the diploid number (2n).
The sex cells (sperms and ova) of a diploid eukaryote cell contain half the number of
chromosomal sets found in the somatic cells and are known as haploid (n) cells. A haploid set of
chromosome is also called genome. The fertilization process restores the diploid number of a
diploid species.
Chemical Structure of Chromosomes
Chemically, the eukaryotic chromosomes are composed of deoxyribonucleic acid (DNA),
ribonucleic acid (RNA), histone and non-histone proteins and certain metallic ions. The histone
proteins have basic properties and have significant role in controlling or regulating the functions
of chromosomal DNA. The non-histone proteins are mostly acidic and have been considered
more important than histones as regulatory molecules. Some non-histone proteins also have
enzymatic activities. The most important enzymatic proteins of chromosomes are
phosphoproteins, DNA polymerase, RNA-polymerase, DPN-pyropbosphorylase, and nucleoside
triphosphatase. The metal ions as Ca+ and Mg+ are supposed to maintain the oragnization of
chromosomes intact.
Molecular Structure of Chromosomes
According to the recent and widely accepted theory of Dupraw (1965, 1970) and Hans
Ris (1967) called unistranded theory, each eukaryotic chromosome is composed of a single,
greatly elongated and highly folded nucleoprotein fibre of 100A 0 thick. This nucleoprotein fibre
in its turn is composed of a single, linear, double stranded DNA molecule which remains
wrapped in equal amounts of histone and non-histone proteins and variable amounts of different
kinds of RNA. Dupraw produced a “folded-fibre Model" to show the ultrastructure of
chromosome.
FIBRE FOLDED MODEL
This model shows a highly folded nucleoprotein fibre in a chromosome and also
suggests that how the nucleoprotein fibre of a chromosome replicates during cell division and
how the nucleoprotein fibre of both chromatids remain held at the centromere by a unreplicated
fibre segment to DNA until the anaphase.
A-B- The folded fibre model of Dupraw for chromosomes in Interphase

1. DNA molecule 2.Protein molecules
3. Chromatids
Material of Chromosomes
The chromatin material of the eukaryotic chromosomes according to its percentage of
DNA, RNA and proteins and consequently due to its, staining property has been classified into
following by classical cytologists:
1. Euchromatin
The euchromatin is the extended form of chromatin and it forms the major, portion of
chromosomes. The euchromatin has special affinity for basic stains and is genetically active
because its component DNA molecule synthesizes RNA molecules only in the extended form
of chromatin.
2. Heterochromatin
The heterochromatin is a condensed intercoiled state of chromatin, containing two to
three times more DNA than euchromatin. However, it is genetically inert as it does not direct
synthesize RNA (i.e., transcription) and protein and is often replicated at a different time from
the rest of the DNA.
Recent molecular biological studies have identified three kinds of heterochromatins,
namely constitutive, facultative and condensed heterochromatin. The constitutive
heterochromatin is present at all times and in the nuclei of virtually all the cells of an organism.
In a interphase nucleus, it tends to clump together to form chromocentre or false nucleoli. In
Drosophila, for example, most pupal, larval and adult cells contain large blocks of constitutive
heterochromatin that lie adjacent to centromeres. Constitutive heterochromatin contain highly
repititive satellite DNA which is late replicating, it fails to replicate until late in the 5-phase and
is then replicated during a brief period just before the G2. The facultative heterochromatin
reflect the existence of a regulatory device designed to adjust the "dosages" of certain genes in
the nucleus
It is originated during the process called facultative heterochromatization : a process in

which a chromosome or a set of chromosome becomes heterochromatic (turned off) in the
cells of one sex, while remaining become heterochromatic (turned on) in the cells of opposite
sex. In other words, it remains indirectly related to, sexual differentiation. The condensed
heterochromatin is deeply staining tightly coiled chromatin which does not resemble with two
other kinds of chromatin, has some specific role in gene regulation and is found in many
interphase nuclei.
Kinds of Chromosomes
The eukaryotic chromosomes have been classified into autosomes and sex
chromosomes. The autosomes have nothing to do with the determination of sex and exceed in
number than sex chromosomes. The sex chromosomes determine the sex of their bearer.
They are usually two in number and are usually of two kinds: X chromosomes and Y
chromosomes.
Special Types of Chromosomes
The eukaryotes besides possessing the usual type of chromosomes in their body cells,
contain some unusual and special types of chromosomes in some body cells or at some
particular stage of their life cycle. The special type of eukaryotic chromosomes are following:
Polytene chromosomes
The nuclei of the salivary gland cells of the larvae of dipterans like Drosophila have
unusually long and wide chromosomes, 100 or 200 times in size of the chromosomes in meiosis
and mitosis of the same species. This is particularly surprising, since the salivary gland cells do
not divide after the glands are formed, yet their chromosomes replicate several times (a process
called endomitosis) and become exceptionally giant-sized to be called polytene or multistranded
chromosomes (discovered by Balbiani (l881) and named by Koller).The endomitosis process
result in the production of 2X chromosomes, where X gives the number of multiplication cycle.
The polytene Chromosomes of the salivary gland cells of D. melanogaster contain 1000
to 2000 chromosomes, which are formed by nine or ten consecutive multiplication cycles and
remain associated parallel to each other. Further, the polytene chromosomes have alternating
dark and light bands along their length. The dark bands are comparable with the chromomeres
of a simple chromosomes and are disc-shaped structures occupying the whole diameter of
chromosome. They contain euchromatin. The light bands or inter bands are fibrillar and
composed of heterochromatin.
A. mRNA
B. Chromosome puff
C. Chromonemata
D. Dark band
E. Inter band
If the polytene chromosomes of dipteran larval salivary glands are examined at several
stages of development; it is seen that specific areas (sets of bands) enlarge or "puff". Such puffs
change location as development proceed, those at specific locations being correlated with
particular developmental stages. This temporal puffing indicates changes in gene activity and
involves several processes such as the accumulation of acidic proteins, despiralization of DNA,
formation of chromonemal loops called Balbiani rings at the lateral sides of dark bands,
synthesis of mRNA (messenger RNA) and storage (accumulation) of newly synthesized mRNA
around the Balbiani rings.
Lampbrush chromosomes
In diplotene stage of meiosis, the yolk rich oocytes of vertebrates contain the nuclei with
many lamp brush shaped chromosomes of exceptionally large sizes. The lampbrush
chromosomes (discovered by Ruckert in 1892) are formed during the active synthesis of mRNA
molecules for the future use by the egg during cleavage when no synthesis of mRNA molecules
is possible due to active involvement of chromosomes in the mitotic cell division.
A lampbrush chromosome contains a main axis whose chromonemal fibres (DNA

molecule) gives out lateral loops throughout its length. The loops produce the mRNA molecules
of different kinds. In a mature egg, as the chromosome , contracts the lateral loops disappear.
B-chromosomes
Many plant (maize, etc.) and animal (such as insects and small mammals) species,
besides having autosomes (A-chromosomes) and sex-chromosomes possess a special
category of chromosomes called B-chromosomes without obvious genetic function. These B-
chromosomes (also called supernumerary chromosomes, accessory chromosomes, accessory
fragments, etc.) usually have a normal structure, are somewhat smaller than the autosomes and
can be predominantly, heterochromatic (many insects, maize, etc. ) or pro-dominantly
euchromatic (rye).
In maize, their number per cell can vary from 0 to 30 and they adversely affect,
development and fertility only when occur, in large amount. In animals, the B-chromosomes
disappear from the non-reproductive (somatic) tissue and are maintained only in the cell-lines
that lead to the reproductive organs. B-chromosomes have negative consequences for the
organism, as they have deleterious effect because of abnormal crossing over during the meiosis
of animals and abnormal nucleus divisions of the gametoophyte plants. In animals, B-
chromosomes occur more frequently in females and the basis is non-disjunction. The non-
disjunction of B-chromosomes of rye plant is found to be caused due to the presence of a

heterochromatic knob at the end of long arm of B-chromosome.
A. Maize B. Rye
Centromere
The origin of the B-chromosomes is uncertain. In some animals they may be derivatives of sex
chromosomes, but this is not the rule. They generally do not show any pairing affinity with the'
A-chromosomes.
Holokinetic chromosomes
The chromosomes of most plants and animals have centromeres that are situated at one
specific position in each chromosome. In a number of animals, especially in insects of the order
Hemiptera and a few, mostly monocotyledonous plants (Juncales, Cyperales), the kinetic
activity is distributed over the entire chromosome and such chromosomes are called Holokinetic
chromosomes (Sybenga, 1972). The term -diffuse centromere bas been used as an alternative
but is not quite logical. In mitotic metaphase, the chromatids of a Holokinetic chromosome orient
parallel in the equator: one chromatid towards one pole the other towards the other pole. This is
also the way they separate in anaphase and they maintain this orientation until they arrive at the
poles. Probably kinetic activity starts at one point and proceed from there on, orienting each unit
to the preceding one.
A-Holokinetic B-Holokinetic C-Meiotic metaphase I bivalent

chromosome at mitotic chromosome at mitotic of a holokinetic chromosome
metaphase anaphase pair
In a number of animals, especially in insects of the order Hemiptera and a few, mostly
monocotyledonous plants (Juncales, Cyperales), the kinetic activity is distributed over the entire
chromosome and such chromosomes ate caned Holokinetic chromosomes (Sybenga, 1972).
The term -diffuse centromere bas been used as an alternative but is not quite logical. In 1966
Flach observed this type of centromere in some primitive DicotyledonS (Ranales:
Myristica.Ascaris and pseudoscorpion Tityus also possess such polycentric chromosomes.
In mitotic metaphase, the chromatids of a Holokinetic chromosome orient parallel in the equator:
one chromatid towards one pole the other towards the other pole. This is also the way they
separate in anaphase and they maintain this orientation until they arrive at the poles. Probably
kinetic activity starts at one point and proceed from there on, orienting each unit to the
preceding one.
Genetic Significance of Chromosomes
The chromosomes are considered as the organs of heredity because of following reasons:
(i) They form the only link between two generations.
(ii) A diploid chromosome set consists of two morphologically similar (except the X and Y sex
chromosomes) sets, one is derived from the mother and another from the father at fertilization.
(iii) The genetic material, DNA or RNA is localized in the chromosome and its contents are
relatively constant from one generation to the next.
(iv) The chromosomes maintain and replicate the genetic informations contained in their DNA
molecule and this information is transcribed at the right time in proper sequence into the specific
types of RNA molecules which directs the synthesis of different types of proteins to form a body
form like the parents.
KARYOTYPE
A karyotype is the characteristic chromosome complement of a eukaryote species. The
preparation and study of karyotypes is part of cytogenetics. The basic number of chromosomes
in the somatic cells of an individual or a species is called the somatic number and is designated
2n. Thus, in humans 2n=46. In the germ-line (the sex cells) the chromosome number is n
(humans: n=23). So, in normal diploid organisms, autosomal chromosomes are present in two
copies. There may, or may not, be sex chromosomes. Polyploid cells have multiple copies of
chromosomes and haploid cells have single copies. The study of whole sets of chromosomes is
sometimes known as karyology. The chromosomes are depicted (by rearranging a
microphotograph) in a standard format known as a karyogram or idiogram: in pairs, ordered by
size and position of centromere for chromosomes of the same size. Karyotypes can be used for
many purposes; such as, to study chromosomal aberrations, cellular function, taxonomic
relationships, and to gather information about past evolutionary events.
Idiogram
Staining
The study of karyotypes is made possible by staining. Usually, a suitable dye is applied
after cells have been arrested during cell division by a solution of colchicine. For humans, white
blood cells are used most frequently because they are easily induced to divide and grow in
tissue culture. Sometimes observations may be made on non-dividing (interphase) cells. The
sex of an unborn fetus can be determined by observation of interphase cells (see amniotic
centesis and Barr body).
Most (but not all) species have a standard karyotype. The normal human karyotypes
contain 22 pairs of autosomal chromosomes and one pair of sex chromosomes. Normal
karyotypes for females contain two X chromosomes and are denoted 46, XX; males have both
an X and a Y chromosome denoted 46,XY. Any variation from the standard karyotype may lead
to developmental abnormalities.
Six different characteristics of karyotypes are usually observed and compared:
1. differences in absolute sizes of chromosomes. Chromosomes can vary in absolute size
by as much as twenty-fold between genera of the same family: Lotus tenuis and Vicia
faba (legumes), both have six pairs of chromosomes (n=6) yet V. faba chromosomes are
many times larger. This feature probably reflects different amounts of DNA duplication.
2. differences in the position of centromeres. This is brought about by translocations.
3. differences in relative size of chromosomes can only be caused by segmental
interchange of unequal lengths.
4. differences in basic number of chromosomes may occur due to successive unequal
translocations which finally remove all the essential genetic material from a
chromosome, permitting its loss without penalty to the organism (the dislocation
hypothesis). Humans have one pair fewer chromosomes than the great apes, but the
genes have been mostly translocated (added) to other chromosomes.
5. differences in number and position of satellites, which (when they occur) are small
bodies attached to a chromosome by a thin thread.
6. differences in degree and distribution of heterochromatic regions. Heterochromatin
stains darker than euchromatin, indicating tighter packing, and mainly consists of
genetically inactive repetitive DNA sequences.
A full account of a karyotype may therefore include the number, type, shape and banding of the
chromosomes, as well as other cytogenetic information.
Variation is often found:
1. between the sexes
2. between the germ-line and soma (between gametes and the rest of the body)
3. between members of a population (chromosome polymorphism)
4. geographical variation between races
5. mosaics or otherwise abnormal individuals
IDEOGRAMS
Ideograms are a schematic representation of chromosomes. They show the relative size
of the chromosomes and their banding patterns. A banding pattern appears when a tightly
coiled chromosome is stained with specific chemical solutions and then viewed under a
microscope. Some parts of the chromosome are stained (G-bands) while others refuse to adopt
the dye (R-bands). The resulting alternating stained parts form a characteristic banding pattern
which can be used to identify a chromosome. The bands can also be used to describe the
location of genes or interspersed elements on a chromosome.
Below is an ideogram of each human chromosome. Next to the known schematic
representation a chromosome was added that was rendered from DNA data. The G-bands,
areas with proportional more A-T base pairs, are normally colored black in schematic
representations. To compare the schematic ideograms the rendered chromosomes, were
colored the A-T bases black and the G-C bases white. Blue areas in the rendered
chromosomes identify bases not known yet.
The results are interesting. When comparing the schematic ideograms with the renderd
chromosomes from our project, a significant conformancy can be seen. Most black areas on the
left side show also black areas on the right side and white areas are also white on the "digital"
chromosomes.
Lecture 14
MITOSIS AND MEIOSIS - THEIR SIGNIFICANCE AND DIFFERENCES BETWEEN THEM

Mitosis occurs only in eukaryotes. Prokaryotes (i.e., archaea and bacteria) divide via
binary fission. Mitosis is the process by which the somatic cells of all multicellular organisms
multiply. Somatic cells are the nonreproductive cells of which an organism is composed.
In addition, plants produce gametes by mitosis. Gametes are sexual reproductive cells,
that is, there are two types, male and female. In sexual reproduction, a male gamete combines
with a female gamete and the resulting, merged cell then divides repeatedly by mitosis to
eventually produce a mature organism. Plants also make asexual reproductive cells called
spores (by meiosis, not mitosis). One spore does not have to combine with another spore for
reproduction to occur. A single spore, produced by meiosis, develops into a mature organism by
mitosis.
Why mitosis?
1. Growth. The number of cells within an organism increases by mitosis and this is the
basis of growth in multicellular organisms.
2. Cell Replacement. Cells are constantly sloughed off, dying and being replaced by new
ones in the skin and digestive tract. When damaged tissues are repaired, the new cells
must be exact copies of the cells being replaced so as to retain normal function of cells.
3. Regeneration. Some animals can regenerate parts of the body, and production of new
cells are achieved by mitosis.
4. Vegetative Reproduction. Some plants produce offspring which are genetically similar to
themselves. These offspring are called clones.
Consequence of mitotic division
• No variation in genetic information
• No variation in chromosome number due to the semi-conservative replication of DNA
and equal distribution of DNA.
• The cell divides once.
• Two identical daughter cells are formed.
Mitosis produces two daughter cells that are identical to the parent cell. If the
parent cell is haploid (N), then the daughter cells will be haploid. If the parent cell is
diploid, the daughter cells will also be diploid.
N→N
2N → 2N
A chromatid is one of the two halves of a replicated chromosome (see diagram at right).
The two chromatids that make up a chromosome are called "sister chromatids," They are joined
at the centromere and are genetically identical because, during interphase, one sister of each
chromatid pair is produced by directly copying the other, pre-existing sister. They therefore
contain identical alleles at all loci. In contrast, two homologous chromosomes (chromosomes
that have the same set of loci in the same order) usually do not have identical alleles at all loci.
They are inherited from different parents and are not direct copies of each other.
The two sister chromatids of each chromosome are segregated into separate cells in
both mitosis and meiosis, but they remain together throughout meiosis I. It is only during the
second meiotic division (meiosis II) that they finally are separated and distributed into separate
cells. As soon as the joined chromatids are separated they are no longer called sisters because
they are no longer connected to each other. Instead they are now called unreplicated
chromosomes.
Stages of Mitosis
The process of mitosis is divided into 6 stages.
Interphase
Prophase
Metaphase
Anaphase
Telophase and
Cytokinasis.
Interphase
At Interphase, there is only one cell, but after cytokinasis there are two identical cells.
Before mitosis can take place, the cell need to store enough energy to drive the chemical
processes during the cell division. During this period of time, there is intense cellular activity.
The cell grows in size. The length of the grow phase varies between a few hours to a few
months. We the cell has stored enough energy, it is ready to divide itself.
The interphase, or growth, period of the cell cycle alternates with the mitotic phase of the
cycle. It's the period when the cell is not undergoing mitotic division. So it is not part of mitosis.
When it begins, the chromosomes (red) have not yet replicated (i.e., each chromosome has a
single chromatid), but by the beginning of prophase replication is complete. Thus, the picture
shown here represents the chromosomes as they are in the first stage of interphase before
replication has occurred, G 1 phase (G 1 stands for first gap). During the next stage, S phase
(synthesis), the chromosomes replicate, and by the beginning of the third, G 2 phase (G 2 stands
for second gap), replication is complete.
Parent Cell (Interphase - G 1 )

In this diagram the chromosomes are shown as if they were visible, simply to show that
they have not replicated. However, during G 1 , S, and G 2 they are not actually visible under a
light microscope, both because they are uncondensed and because they are still enclosed in
the nuclear membrane.
Prophase
Prophase is the first of the four stages of mitosis. Early during this stage the
chromosomes (shown in red in the diagram) become visible with a light microscope as they
condense (i.e., as they shorten, coil, and thicken). Also, the spindle (yellow strands) begins to
extend outward from each of two centers of extension. This starlike configuration is called an
aster. In animal cells one pair of centrioles (represented by the yellow cylinders in the diagram)
is present at each centers of extension. As prophase progresses, the nuclear membrane (white)
begins to break up and disappear. Each chromosome has been duplicated and so is composed
of two sister chromatids containing identical genetic information.
METAPHASE
During this, the second stage of mitosis, the chromosomes line up in the middle of
the cell, halfway between the centrioles on an imaginary plane called the "metaphase plate"
The spindle fibers (yellow strands) attach to the centromeres (shown as yellow ovals).
ANAPHASE
During the third of the four stages of mitosis, the two chromatids of each chromosome
are pulled apart by the spindle and dragged toward opposite poles of the cell (i.e., toward the
opposite centrioles). The arms of each chromosome can be seen dragging behind as the it is
dragged along by its centromere.
Note: To be called a chromatid, a chromatid must be attached to its sister. When the
chromosomes divide at the beginning of anaphase, the sister chromatids are no longer sisters
because they are no longer connected. Once they are separated, they are called unreplicated
chromosomes. Unreplicated chromosomes will come up again in the Interphase section below.
TELOPHASE
During the last of the four stages of mitosis, telophase, the chromosomes have reached
the poles and the nuclear membrane begins to appear. During telophase in animal cells, a
cleavage furrow appears (see photo below). By the end of this stage of mitosis, the cell has
divided in two along the plane defined by the furrow. In most plants, instead of a cleavage
furrow, a "cell plate" forms, dividing the cell into two daughter cells.
CYTOKINESIS
While cytokinesis is one of the steps in the cell cycle, it is not one of the phases of
mitosis. It is the division of the cytoplasm, as opposed to karyokinesis, which is division of the
nucleus (the cytoplasm is all of the contents of the cell outside the nucleus). Division of the
cytoplasm occurs in both mitosis and meiosis.
After mitosis, the cell returns to interphase, which recall, is the growth stage of the cell
cycle between successive mitoses (interphase is the stage during which DNA synthesis, or
replication, occurs). Cytokinesis and karyokinesis are now complete and there are two daughter
cells. The nuclear membrane has reformed. The chromosomes have decondensed once again
and are now re-enclosed in an intact nuclear membrane.
Significance of mitosis
The significance of mitosis is its ability to produce daughter cells which are exactly the same a
the parent cell. It is important for three reasons:
1. Growth
If a tissue wants to get bigger by growth needs new cells that are identical to the existing
ones. Cells division must therefore be by mitosis.
2. Repair
Damaged cells have to be replaced by exact copies of the organism so that it repairs the
tissues to their former condition. Mitosis is the means by which this is achieved.
3. Asexual reproduction
If a species is good at colonizing a habitat, there might be no point, in producing offspring
which are different from the parents, because they might be less effective at survival.
Therefore it might be better, in the short term, to make a colony which is similar to the
parents. In simple animals and most plants this is achieved by mitotic division.
MEIOSIS
When does Meiosis Occur?
Meiosis occurs only in eukaryotes. Meiosis takes place at some point in the life cycle of
the typical sexual organism because, by reducing the chromosome number by one half, it
compensates for the doubling of the chromosome number caused by fertilization. In animals,
meiosis occurs during the production of gametes (sperm and eggs). In plants, it takes place
when spores are produced (plant gametes are produced by mitosis). Prokaryotes (i.e., archaea
and bacteria) reproduce via binary fission.
The names of the eight stages of meiosis are: prophase I, metaphase I, anaphase I,
telophase I, prophase II, metaphase II, anaphase II, and telophase II. The first four steps (the
ones ending in "I") are the phases of the first meiotic division, meiosis I. The last four (those
ending in "II") are those of the second meiotic division, meiosis II.
Prophase I
Prophase I is the first stage of meiosis. During this phase, the chromosomes (shown in
red in the diagram, below right) become visible as they shorten, coil, and thicken. Also, the
spindle (yellow strands in diagram) begins to extend outward from two centers of expansion. In
animal cells a pair of centrioles can be seen in each of these centers (plants lack centrioles).
The nuclear membrane (shown in white) breaks up and disappears. Each chromosome is
composed of two sister chromatids containing identical genetic information. The information is
the same because one sister chromatid is produced by copying the other. The sister chromatids
are joined by a centromere (orange).
The two members of each chromosome pair are called homologous chromosomes or
simply homologs. During prophase I they join along their lengths (i.e., they synapse) to form a
tetrad (or bivalent). Each of the two tetrads shown in the drawing, then, is represented as two x-
shaped chromosomes aligned along their lengths and connected with each other. This is a
simplified diagram, the actual situation is more complicated:
Prophase I is by far the most complicated phase of meiosis. It is much longer in meiosis
than in mitosis. During this stage, homologous chromosomes join (synapse) along their lengths
and exchange DNA. Prophase I is itself divided into the five substages which are explained and
diagrammed below.
Leptotene
The chromosomes have appeared within the nuclear membrane (shown in the diagram
at right as a tan circle with a brown border), but are not yet fully condensed. In the diagram the
two chromosomes of paternal origin are indicated in red, those of maternal origin, in blue. Each
is a thin thread of DNA (lepto- is Greek for thin and -tene is Greek for ribbon or band) along
which clearly defined beads of local coiling (chromomeres) can be seen. The chromosomes,
while they have this threadlike form, are called chromatonemata (sing. chromonema; -nema is
Greek for thread). The chromosomes appear single because the sister chromatids are still so
tightly bound to each other that they cannot be separately seen. During this stage both ends
(telomeres) of each chromosome are turned toward, and probably attached to, the same region
of the nuclear membrane. Leptotene is also known as
(1) leptonema; and as
(2) the bouquet stage because all the telomeres tend to contact the nuclear membrane in one
spot so that the looped chromosomes balloon out from that point like flower petals.
Zygotene (also known as zygonema)

During this stage, homologous chromosomes (or homologs for short) begin to unite
(synapse) by coming into approximate alignment (zygo- is Greek for union, fusing, or yoking).
Synapsis, the process of fusion that occurs between homologous chromosomes, begins at
various points along the chromosomes and extends outward, zipper-fashion, until complete.
When synapsis is finished, the fused homologous chromosomes look like single chromosomes
under the light microscope. These chromosomes that look single, but that are actually double,
are called bivalents. The interface where two homologous chromosomes ("homologs" for short)
unite is called a synaptonemal complex, which can be seen under an electron microcope
In the diagram of early zygotene (above right), the regions where the paternal and
maternal homologs have fused is shown in purple. In the next diagram, representing late
zygotene, both homolog pairs have fused over their entire lengths (so they are shown entirely in
purple).
Once the the homolog pairs synapse they are called tetrads (each has four chromatids;
tetra is Greek for four) or bivalents. Bivalent is the preferred term, but tetrad is, nonetheless, the
word more commonly used in most introductory biology classes. Bivalent is the better choice
because there are equivalent names for other situations. For example, an unfused homolog is
called a univalent. Three fused homologs, a common situation in plants, is called a trivalent, etc.
Pachytene
(also known as pachynema). During pachytene the two sister chromatids of each
chromosome separate from each other. This makes the chromosomes look thicker (pachy- is
Greek for thick). Homologs are still paired at this point. Non-sister chromatids remain in contact
throughout pachytene and a kind of localized breakage of the DNA occurs, which is followed by
exchanges of DNA between them. This process is called "crossing over." Crossing over
produces "cross-over chromatids" each composed of distinct blocks of DNA, some blocks
derived from the mother, others from the father.
Diplotene (also known as diplonema)

At the beginning of this stage each chromatid of each chromosome is still fused to a
chromatid of that chromosome/s homolog (recall that sister chromatids are already separate at
this point). As diplotene progresses, these initially fused non-sister chromatids begin to separate
from each other. However, they cannot separate completely because they are still connected by
two strands of DNA at each of the points where exchanges took place. At each cross-over site,
the two strands form an x-shaped structure called a chiasma (pl. chiasmata). The chiasmata
then begin moving toward the ends of the chromatids. This process of sliding toward the ends is
known as terminalization.
In oocytes, a special, extremely prolonged form of diplotene occurs called dictyotene. The
primary oocyte undergoes the first three of the substages of prophase I (leptotene, zygotene,
and pachytene) during late fetal life. The process is then suspended during diplotene until
puberty or thereafter. Therefore, in oocytes dictyotene (and consequently prophase I) can last
months or even years, depending on the type of organism in question.
Diakinesis
During this, the last stage of Prophase I, the nucleolus disappears, terminalization
reaches completion, and the chromosomes coil tightly, and so become shorter and thicker. The
nuclear envelope begins to disappear.
(click here to see more details). Note that the two homologs making up a chromosome pair are
not expected to be genetically identical, as in the case of sister chromatids, because they are
not direct copies of each other and they are inherited from different parents.
When the chromosomes synapse during prophase, each gene in each chromosome is
brought into contact with the same gene on that chromosome's homolog. During this process of
synapsis, the two homologs of each pair exchange segments of DNA in a process known as
crossing over. As a result, the gene combinations on each chromosome can be changed. (For
example, suppose one homolog initially contained genes for brown eyes and brown hair. After
crossing over, it could contain genes for blue eyes and brown hair, where the gene for blue eyes
was taken from its homolog.
While prophase I is proceeding, the spindle's two centers of expansion move to the
opposite ends of the cell (i.e., to the "poles") and the spindle lengthens and extends toward the
"metaphase plate," an imaginary plane defining the middle plane of the cell, halfway between
the centriole pairs. The tetrads also move toward the metaphase plate.
METAPHASE I
In the second phase of the first meiotic division, metaphase I, the tetrads align on the
"metaphase plate," halfway between the poles of the cell. Next, the spindle fibers attach to the
centromeres of each chromosome. Both spindle fiber attachment points (kinetochores) of each
sister chromatid pair are turned toward the same pole. As a result, both kinetochores attach to
spindle fibers from the same pole. This is a major difference between meiosis and mitosis. It
causes the two members of each chromosome pair to be separated from each other during the
next stage of meiosis, anaphase I (in mitotic metaphase, the two kinetochores of each sister
chromatid pair attach to spindle fibers from opposite poles, so each chromatid separates from
its sister during anaphase).
ANAPHASE I
In the third stage, anaphase I, the cell lengthens as it begins the process of division.
Homologs of each chromosome pair move toward opposite poles, drawn by the microtubles of
the spindle apparatus (this contrasts with mitosis, where the sister chromatids from each
homolog separate and move toward opposite poles).
TELOPHASE I
In this, the fourth stage of meiosis, the chromosomes reach the poles. At each pole, now,
there is a complete haploid set of chromosomes (but each chromosome still has two sister
chromatids). During telophase I, a cleavage furrow appears. By the end of this stage the cell
has divided in two along the plane defined by the furrow. This separation of the cytoplasm is
called cytokinesis. In some organisms the nuclear membrane reappears briefly at this point (this
intermediate stage is called interkinesis), but in others the daughter cells begin immediately to
prepare for the second meiotic division.
CYTOKINESIS
Cytokinesis is not one of the stages or phases of meiosis or mitosis. It is the process of
division that the cytoplasm undergoes when it is distributed into daughter cells (as opposed to
karyokinesis, which is division of the nucleus). The cytoplasm is all of the contents of the cell
other than the nucleus. The cytoplasm lies outside the nucleus and is bounded by the plasma
membrane. The liquid portion of cytoplasm is called cytosol.
Interkinesis
In many organisms, after meiosis I, the daughter cells begin immediately to prepare for
the second meiotic division. In others, however, the nuclear membrane reappears between
telophase I and prophase II, and there is a period of rest. This period, during which the
membrane is again visible, is called interkinesis. Each chromosome is still composed of two
chromatids.
PROPHASE II
Prophase II begins with the two daughter cells produced by the first meiotic division. As
in Prophase I, the chromosomes are condensed and not yet attached to the spindle apparatus.
If there was an interkinesis, then the nuclear membranes begin to break down again during this
stage. The centrioles have replicated and are moving toward the poles.
METAPHASE II
In metaphase II the chromosomes move to the equator ("metaphase plate") of each of
the two daughter cells produced by the first meiotic division. This time, unlike metaphase I, the
two kinetochores of each centromere attach to spindle fibers from opposite poles
ANAPHASE II
During the seventh stage of meiosis, anaphase II (the third stage of the second division),
the sister chromatids of each chromosome separate and move toward opposite poles.
TELOPHASE II
During telophase II, the sister chromatids reach opposite poles, cytokinesis occurs, the
two cells produced by meiosis I divide to form four haploid daughter cells, and the nuclear
membranes (white in the diagram) reform.
Significance of Meiosis
The long term survival of a species depends on its ability to adapt to a changing
environment. To do this the offspring need to be different from their parents and each other.
These are three ways in which variety occurs because of meiosis.
a. Production and fusion of haploid gametes
The variety of offspring is increased by mixing the genotype of one parent with that of the
other. It involves the production of special sex cells, called gametes, which fuse together to
produce a new organism. Each gamete contains half the number of chromosomes of the adult.
It is important that meiosis, which halves the number of chromosomes in daughter cells,
happens at some stage in the life cycle of a sexually reproducing organism. Therefore Meiosis
is important in order for variety in organisms, and allowing them to evolve.
b. The creation of genetic variety by the random distribution of chromosomes during
metaphase 1
When the pairs of homologous chromosomes arrange themselves on the equator of the
spindle during metaphase 1 of meiosis, they do it randomly. Even though each one of the pair
determines the same general features, they’re detail of the feature is different. The randomness
of this distribution and independent assortment of these chromosomes produces new genetic
combinations.
c. The creation of genetic variety by crossing over between homologous
chromosomes
During prophase 1 of meiosis, equal portions of homologous chromosomes may be swapped.
In this way new genetic combinations are made and linked genes separated. The variety which
meiosis brings vital for to the process of evolution. By providing a varied stock of individuals it
allows the natural selection of those best suited to the existing conditions and makes sure that
species constantly change and adapt when these conditions change. This is the main biological
significance of meiosis.
Meiosis Mitosis
A type of cellular reproduction in A process of asexual reproduction
which the number of in which the cell divides in two
chromosomes are reduced by producing a replica (exact copy)
Definition
half through the separation of with an equal number of
homologous chromosomes in a chromosomes (One cell becomes
diploid cell. two)
Cellular Reproduction (cell division

Growth, Repair, asexual
Function during which the cell nucleus
Reproduction
divides)
Type of
Sexual Asexual
Reproduction
Discovered
Oscar Hertwig Walther Flemming
by
Makes everything other than sex
Creates Sex cells only
cells (Somatic Cells)
Occurs in Humans, animals, plants all organisms
Produces four haploid daughter cells two diploid daughter cells
Genetically different identical
Steps Proceded by Interphase Proceded by Interphase
Chromosome Reduced by half (homologous
Remains the same
Number pairs seperate)
Number of 2 nuclear and cytoplasmic 1 division of the nucleus in
Divisons divisions Cytokinesis
Lecture 15
DNA AND IT’ S STRUCTURE, FUNCTION, TYPES, MODES OF

REPLICATION AND REPAIR
The discovery that DNA is the prime genetic molecule, carrying all
the hereditary information within chromosomes, immediately had its
attention focused on its structure. It was hoped that knowledge of the
structure would reveal how DNA carries the genetic messages that are
replicated when chromosomes divide to produce two identical copies of
themselves. During the late 1940s and early 1950s, several research
groups in the United States and in Europe engaged in serious efforts—
both cooperative and rival—to understand how the atoms of DNA are
linked together by covalent bonds and how the resulting molecules are
arranged in three-dimensional space. Not surprisingly, it was feared that
DNA might have very complicated and perhaps bizarre structures that
differed radically from one gene to another. Great relief, if not general
elation, was thus expressed when the fundamental DNA structure was
found to be the double helix. It told us that all genes have roughly the
same three-dimensional form and that the differences between two genes
reside in the order and number of their four nucleotide building blocks
along the complementary strands.
What is DNA?
The work of many scientists paved the way for the exploration of
DNA. Way back in 1868, almost a century before the Nobel Prize was
awarded to Watson, Crick and Wilkins, a young Swiss physician named
Friedrich Miescher, isolated something no one had ever seen before from
the nuclei of cells. He called the compound "nuclein." This is today called
nucleic acid, the "NA" in DNA (deoxyribo-nucleic-acid) and RNA (ribo-
nucleic-acid).
Two years earlier, the Czech monk Gregor Mendel, had finished a
series of experiments with peas. His observations turned out to be closely
connected to the finding of nuclein. Mendel was able to show that certain
traits in the peas, such as their shape or colour, were inherited in different
packages. These packages are what we now call genes.
For a long time the connection between nucleic acid and genes was
not known. But in 1944 the American scientist Oswald Avery managed to
transfer the ability to cause disease from one strain of bacteria to another.
But not only that: the previously harmless bacteria could also pass the
trait along to the next generation. What Avery had moved was nucleic
acid. This proved that genes were made up of nucleic acid.
Solving the Puzzle
In the late 1940's, the members of the scientific community were
aware that DNA was most likely the molecule of life, even though many
were skeptical since it was so "simple". They also knew that DNA included
different amounts of the four bases adenine, thymine, guanine and
cytosine (usually abbreviated A, T, G and C), but nobody had the slightest
idea of what the molecule might look like.
In order to solve the elusive structure of DNA, a couple of distinct
pieces of information needed to be put together. One was that the
phosphate backbone was on the outside with bases on the inside; another
that the molecule was a double helix. It was also important to figure out
that the two strands run in opposite directions and that the molecule had
a specific base pairing.
Watson and Crick
In 1951, the then 23-year old biologist James Watson travelled
from the United States to work with Francis Crick, an English physicist at
the University of Cambridge. Crick was already using the process of X-ray
crystallography to study the structure of protein molecules. Together,
Watson and Crick used X-ray crystallography data, produced by Rosalind
Franklin and Maurice Wilkins at King's College in London, to decipher
DNA's structure.
This is what they already knew from the work of many scientists,
about the DNA molecule:
1. DNA is made up of subunits which scientists called nucleotides.
2. Each nucleotide is made up of a sugar, a phosphate and a base.
3. There are 4 different bases in a DNA molecule:
adenine (a purine)
cytosine (a pyrimidine)
guanine (a purine)
thymine (a pyrimidine)
4. The number of purine bases equals the number of pyrimidine bases
5. The number of adenine bases equals the number of thymine bases
6. The number of guanine bases equals the number of cytosine bases

7. The basic structure of the DNA molecule is helical, with the bases
being stacked on top of each other
Components of DNA
DNA is a polymer. The monomer units of DNA are nucleotides, and
the polymer is known as a "polynucleotide". Each nucleotide consists of a
5-carbon sugar (deoxyribose), a nitrogen containing base attached to the
sugar, and a phosphate group. There are four different types of
nucleotides found in DNA, differing only in the nitrogenous base. The four
nucleotides are given one letter abbreviations as shorthand for the four
bases.
• A is for adenine
• G is for guanine
• C is for cytosine
• T is for thymine
Purine Bases
Adenine and guanine are purines. Purines are the larger of the two
types of bases found in DNA. Structures are shown below:
The 9 atoms that make up the fused rings (5 carbon, 4 nitrogen) are
numbered 1-9. All ring atoms lie in the same plane.
Pyrimidine Bases
Cytosine and thymine are pyrimidines. The 6 stoms (4 carbon, 2
nitrogen) are numbered 1-6. Like purines, all pyrimidine ring atoms lie in
the same plane.
Deoxyribose Sugar
The deoxyribose sugar of the DNA backbone has 5 carbons and 3
oxygens. The carbon atoms are numbered 1', 2', 3', 4', and 5' to
distinguish from the numbering of the atoms of the purine and pyrmidine
rings. The hydroxyl groups on the 5'- and 3'- carbons link to the
phosphate groups to form the DNA backbone. Deoxyribose lacks an
hydroxyl group at the 2'-position when compared to ribose, the sugar
component of RNA.
Nucleosides
A nucleoside is one of the four DNA bases covalently attached to
the C1' position of a sugar. The sugar in deoxynucleosides is 2'-
deoxyribose. The sugar in ribonucleosides is ribose. Nucleosides differ
from nucleotides in that they lack phosphate groups. The four different
nucleosides of DNA are deoxyadenosine (dA), deoxyguanosine (dG),
deoxycytosine (dC), and (deoxy)thymidine (dT, or T).
In dA and dG, there is an "N-glycoside" bond between the sugar C1' and
N9 of the purine.
Nucleotides
A nucleotide is a nucleoside with one or more phosphate groups covalently
attached to the 3'- and/or 5'-hydroxyl group(s).
DNA Backbone
The DNA backbone is a polymer with an alternating sugar-
phosphate sequence. The deoxyribose sugars are joined at both the 3'-
hydroxyl and 5'-hydroxyl groups to phosphate groups in ester links, also
known as "phosphodiester" bonds.
Example of DNA Backbone: 5'-d (CGAAT)
Features of the 5'-d(CGAAT) structure:

• Alternating backbone of deoxyribose and phosphodiester groups
• Chain has a direction (known as polarity), 5'- to 3'- from top to

bottom
• Oxygens (red atoms) of phosphates are polar and negatively
charged
• A, G, C, and T bases can extend away from chain, and stack atop
each other
• Bases are hydrophobic
DNA Double Helix
DNA is a normally double stranded macromolecule. Two
polynucleotide chains, held together by weak thermodynamic forces, form
a DNA molecule.
Structure of DNA Double Helix
Features of the DNA Double Helix

• Two DNA strands form a helical spiral, winding around a helix axis
in a right-handed spiral
• The two polynucleotide chains run in opposite directions
• The sugar-phosphate backbones of the two DNA strands wind
around the helix axis like the railing of a sprial staircase
• The bases of the individual nucleotides are on the inside of the

helix, stacked on top of each other like the steps of a spiral
staircase.
The Double Helix

The double helix of DNA has these features:
• It contains two polynucleotide strands wound around each other.
• The backbone of each consists of alternating deoxyribose and
phosphate groups.
• The phosphate group bonded to the 5' carbon atom of one
deoxyribose is covalently bonded to the 3' carbon of the next.
• The two strands are "antiparallel"; that is, one strand runs 5′ to 3′
while the other runs 3′ to 5′.
• The DNA strands are assembled in the 5′ to 3′ direction and, by
convention, we "read" them the same way.
• The purine or pyrimidine attached to each deoxyribose projects in
toward the axis of the helix.
• Each base forms hydrogen bonds with the one directly opposite it,
forming base pairs (also called nucleotide pairs).
• 3.4 Å separate the planes in which adjacent base pairs are located.
• The double helix makes a complete turn in just over 10 nucleotide
pairs, so each turn takes a little more (35.7 Å to be exact) than the
34 Å shown in the diagram.
• There is an average of 25 hydrogen bonds within each complete
turn of the double helix providing a stability of binding about as
strong as what a covalent bond would provide.
• The diameter of the helix is 20 Å.
• The helix can be virtually any length; when fully stretched, some
DNA molecules are as much as 5 cm (2 inches!) long.
• The path taken by the two backbones forms a major (wider) groove
(from "34 A" to the top of the arrow) and a minor (narrower)
groove (the one below).
Nucleic acids (DNA and RNA) are the polymers i.e. long chain
compounds. The molecular structure of DNA has two aspects
1) its chemical sub units and
2) the way in which these chemical sub units are arranged to form a long
chain molecule.
The second aspect is very significant as the accepted DNA model
should be such that it explains biochemically the various aspects
(function) of a gene such as stability to metabolic and external agents,
the capacity for replication (self duplication) the capacity to store vast
hereditary information in coded form and the capacity to express the
phenotypes they control.
FUNCTIONS OF DNA
DNA carries the genetic information of a cell and consists of
thousands of genes. Each gene serves as a recipe on how to build a
protein molecule. Proteins perform important tasks for the cell functions
or serve as building blocks. The flow of information from the genes
determines the protein composition and thereby the functions of the cell.
The DNA is situated in the nucleus, organized into chromosomes.
Every cell must contain the genetic information and the DNA is therefore
duplicated before a cell divides (replication). When proteins are needed,
the corresponding genes are transcribed into RNA (transcription). The
RNA is first processed so that non-coding parts are removed
(processing) and is then transported out of the nucleus (transport).
Outside the nucleus, the proteins are built based upon the code in the
RNA (translation).
Types of DNA
DNA can be classified in various ways based on 1. number of base pair per
turn. 2. coiling pattern, 3. location 4. structure, 5. nucleotide sequence
and 6. number of strands.
1. Number of base per turn. Depending upon the nucleotide base per
turn of the helix, tilt of the base pair and humidity of the sample, the DNA
can be observed in four different forms namely A,B, C and D.
2. Coiling pattern. On the basis of coiling pattern of the helix DNA is of
two types viz right handed and left handed. Most of the DNA molecules
are right handed i.e. coiling of helix is in the right direction. It is also
called positive coiling. All the four forms of DNA viz A, B, C and D are right
handed. The Z DNA has left handed double helical structure. This DNA is
considered to be associated with gene regulation.
3. Location. Based on the location in the cell DNA is of three types. Viz.,
chromosomal DNA cytoplasm DNA and promiscuous DNA. Chromosomal
DNA is found in chromosomes. And are called as chromosomal DNA or
nuclear DNA. Cytoplasmic DNA is found in the cytoplasm especially in
mitochondria and chloroplasts. Such DNA plays an important role in
cytoplasmic inheritance and has circular structure. Promiscuous DNA.
Some DNA segments with common base sequence are found in the
chloroplasts, mitochondria and nucleus. This suggests that some DNA
sequences move from one organelle to other. Such DNA is referred to as
promiscuous DNA.
4. Structure of RNA: It contains ribose sugar, nitrogen bases and
phosphate group. The nitrogen bases include adenine, guanine, cytosine
and uracil. In DNA thymine is present in place of uracil and deoxyribose
sugar is found in place of ribose sugar. In RNA, the pairing occurs
between adenine and uracil. It has usually single strand. However, some
viruses have double stranded RNA.
The DNA molecule that Watson and Crick described was in the B
form. It is now known that DNA can exist in several other forms. The
primary difference between the forms is the direction that the helix
spirals.
A, B, C = right-handed helix Z = left-handed helix (found in vitro under

high salt)
B is the major form that is found in the cell. Z-DNA was initially
found only under high salt conditions, but the cellular environment is
actually a low-salt environment. The question then is whether type Z exist
under cellular conditions. Several features have been discovered that can
stablize Z-DNA under in a low salt environment.
Differences between DNA and RNA
S. No Particulars DNA RNA

1. Strands Usually two, rarely one Usually one, rarely two
2. Sugar Deoxyribose Ribose
3. Base Adenine guanine Adenine guanines

cytosine cytosine
and thymine
4. Pairing AT and GC AU and GC
5. Location Mostly in chromosomes In chromosomes and
some in mitochondria ribosomes
and
chloroplasts
MODES OF REPLICATION
There are three possible modes of DNA replication:
(1) Dispersive
(2) Conservative
(3) Semiconservative
1. In dispersive replication, the old DNA molecule would break into
several pieces, each fragment would replicate and the old and new
sesgments would recombine randomly to yield progeny DNA molecules.
Each progeny molecule would have both old and new segments along its
length.
2. According to the conservative scheme, the two newly synthesized
strands ( following the replication of a DNA molecule) would associate to
form one double helix, while the two old strands would remain together as
one double helix.
3. In contrast, in the semi conservative mode of DNA replication, each
newly synthesized strand would remain associated with the old strand
against which it was synthesized. Thus each progeny DNA molecule would
consist of one old and one newly synthesized strand.
Semi Conservative Replication
The semi conservative mode of DNA replication was postulated by
Watson and Crick along with the double helix model of DNA. The main
features of this mode of DNA replication are as follows:
1. A progressive separation of the two strands of a DNA molecule.
2. Complementary base pairing of the bases located in the single stranded
regions thus produced with the appropriate free deoxyribonulceotides.
3. Formation of phosphodiester linkages between the neighbouring

deoxyribonucleotides that have base paired with the single stranded
regions, thereby producing regions the new strand.
4. This ensures that the base sequence of the new strands are strictly
complementary top those of the old strands.
5. The base sequence of a newly synthesized strand is dictated by the
base sequence of the old strand, since the old strand serves as a template
or lould for the synthesis of the new strand.
DNA Replication
It is proposed by Watson and Crick. According to this method, both
the strands of parental DNA separate from one another. Each old strand
synthesizes a new strand. Thus, each of the two resulting DNA has one
parental and one new strand. This method of DNA replication is
universally accepted because there are several evidences in support of
semi conservative method and it consists of several steps.
1. Initiation of Replication DNA replication starts at a specific point on
the chromosome. This unique site is known as origin. The site of initiation
differs from organism to organism. Sometime replication starts with an
incision made by an incision enzyme known as endonuclease.
2. Unwinding of strands. The two stands of DNA double helix unwind.
The opening of DNA stands take’s places with the help of DNA unwinding
protein.
3. Formation of RNA Primer. Synthesis of RNA primer is essential for
initiation of DNA synthesis RNA primer is synthesized by the DNA template
near the origin with the help of a special type of RNA polymerase.
4. Synthesis of DNA on primer. After formation of RNA primer, DNA
synthesis starts on the RNA primer. Deoxyribose nucleotides are added to
the 3e end position of RNA primer. The main DNA strand is synthesized on
the DNA template with help of DNA polymerase. The DNA synthesis takes
place in short pieces. Which are known as Okazaki fragments.
5. Removal of RNA Primer: DNA polymerase degrades the RNA primer
1. This enzyme also catalyzes the synthesis of short DNA segment to
replace the prime. The newly synthesized segment is joined to the main
DNA strand with the help of DNA ligase enzyme.
6. Union of Okazaki Fragments. The discontinuous fragment of Okazaki

is joined to make continuous strands. The union of Okazaki fragments
takes place with the help of a joining enzyme called polynucleotide ligase.
The replication may take place either in one direction or in both the
directions from the point of origin.
Evidence for semi conservation replication
Various experiments have demonstrated the semi-conservative
mode of DNA replication. Now it is universally accepted that DNA
replicates in a semi-conservative manner. There are three important
experiments, which support that DNA replication is semi-conservative.
These include (1) Meselson and Stahl experiment (2) Cairns experiment
and (3) Taylor.s experiment.
Taylor.s experiment: Taylor (1969) conducted his experiments with root
tip cells of vicia faba. He treated root tips with radioactive thymidine to
label the DNA. The root tips were grown in the normal medium. In the
first generation both chromatids were labeled. In the second generation of
cell division, one chromatid of each chromosome was labeled and the
other one was normal. This demonstrated semi conservative mode of
chromosome replication. The DNA replication is associated with
chromosome replication.
Enzymes involved in DNA / RNA replication
DNA replication involves several proteins and enzymes, which
together form the multienzymes complex, rep0lication apparatus or
replisome. In E coli at lest two dozen gene products are involved in DNA
replication. Many of these protein were first identified through studies of
mutants e.g. Genes dna E, dna N, dna x etc of E colic code for the four of
the seven polypeptides of the complete DNA polymerase III enzyme, and
DNA G specifies the primase enzyme. Some enzymes like ligase, DNA
polymerase 1 etc were discovered biochemically.
DNA repair systems
Damages to the genetic material, i.e., DNA are taken care of by the DNA
repair systems. The various damages to DNA may be grouped into the
following two types:
(1) Single base changes: Such changes affect a single base of a DNA
molecule they do not produce structural distortions and do not affect
either replication or transcription of the affected molecules. These

changes ar represented by the conversion of one base into another, eg;
deamination of 5 methylcytosine results in thymine and by the covalent
addition of a small group to a base which affects its pairing behavior. As a
result, the affected base does not pair properly with its partner base.
(2) Structural distortations: These changes generally adversely affect
the replication and or transcription of the affected DNA molecule. They are
represented by a single strand nick, removal of a base, covalent links
between bases in the same or in the opposite strands (eg) Pyrimidine
dimmers and addition of a bulky adduct to a base which may distort the
configuration of the double helix.
The repair systems recognize a variety of changes in DNA to initiate
action. Each cell possesses several repair systems in order to be able to
deal with the various types of DNA damage; these systems may be
grouped into the following general categories
1. Direct repair
2. Excision repair
3. Mismatch repair
4. Tolerance systems
5. Retrieval systems
1. Direct repair of DNA
The reversal or simple removal of the damage to the DNA is known
as direct repair, eg., removal of the covalent bonds between the two 4
and two 5 carbons of the two thymine residues participating in the
formation of thymine dimmers. Thymine dimers are generally formed due
to UV radiation and interfere with replication and transcription. A specific
enzyme mediates the splitting of the covalent bonds between the two T
residues, which specifically recognizes to thymine dimmers. The enzyme
can bind to the thymine dimmers in the dark, but requires the energy
from blue light for removal of the covalent bonds between the T residues;
that is why this process is known as photoreactivation. The direct repair
system is wide spread in nature and is especially important in plants.
2. Excision repair
In this repair pathway, the damaged or mispaired segment of the
DNA strand is exercised and new stretch of DNA is synthesized in its
place. The various excision repair systems vary in their specificity. The
repair process consists of the following steps:
a. Recognition and incision: The damaged section of a strand
recognized by an endonuclease; this enzyme then cuts the affected strand
on both the sides of damage.
b. Excision: After the incision, a 5’ to 3 ‘ exonulcease digests away the
damage/ mispaired section; this generates a single stranded region in the
DNA double helix.
c. Synthesis: In this step, the single stranded region produced by
excision serves as a template for a DNA polymerase which synthesis the
replacement for the excised segment. DNA ligase then seals the nick that
remains after the synthesis of the replacement for the excised section.
3. Mismatch repair: When single bases in the DNA are mismatched,
either due to alterations in the existing bases or due to errors during
replication, structural distortions result in the DNA double helix.
4. Tolerance systems: These systems deal with the damages that block
normal replication at the damaged sites possibly by permitting the
replication of the damaged sites possibly with a high frequency of errors.
These systems may be particularly important in the eukaryotes where the
genome size is very large and hence a complete repair of the damage is
rather unlikely.
5. Retrieval systems: These systems are also known as post replication
repair or recombination repair.
Lecture 16
RNA AND ITS STRUCTURE, FUNCTION AND TYPES
With the discovery of the molecular structure of the DNA double helix in
1953, researchers turned to the structure of ribonucleic acid (RNA) as the next
critical puzzle to be solved on the road to understanding the molecular basis of
life. Ribonucleic acid (RNA) is a type of molecule that consists of a long chain of
nucleotide units. Each nucleotide consists of a nitrogenous base, a ribose sugar,
and a phosphate. RNA is very similar to DNA, but differs in a few important
structural details: in the cell, RNA is usually single-stranded, while DNA is usually
double-stranded; RNA nucleotides contain ribose while DNA contains deoxyribose
(a type of ribose that lacks one oxygen atom); and RNA has the base uracil
rather than thymine that is present in DNA.
RNA is transcribed from DNA by enzymes called RNA polymerases and is
generally further processed by other enzymes. RNA is central to the synthesis of
proteins. Here, a type of RNA called messenger RNA carries information from DNA
to structures called ribosomes. These ribosomes are made from proteins and
ribosomal RNAs, which come together to form a molecular machine that can read
messenger RNAs and translate the information they carry into proteins. There are
many RNAs with other roles – in particular regulating which genes are expressed,
but also as the genomes of most viruses.
Ribose Nucleic Acids
Most cellular RNA is single stranded, although some viruses have double
stranded RNA. The single RNA strand is folded upon itself, either entirely or in
certain regions. In the folded region a majority of the bases are complementary and
are joined by hydrogen bonds. This helps in the stability of the molecule. In the
unfolded region the bases have no complements. Because of this RNA does not
have the purine, pyrimidine equality that is found in DNA.
RNA also differs from DNA in having ribose as the sugar instead of
deoxyribose. The common nitrogenous bases of RNA are adenine, guanine, cytosine
and uracil. Thus the pyrimidine uracil substitutes thymine of DNA. In regions where
purine pyrimidine pairing takes place, adenine pairs with uracil and guanine with
cytosine. In addition to the four bases mentioned above, RNA also has some
unusual bases.
CHEMICAL STRUCTURE OF RNA
An important structural feature of RNA that distinguishes it from DNA is the

presence of a hydroxyl group at the 2' position of the ribose sugar. The presence
of this functional group causes the helix to adopt the A-form geometry rather
than the B-form most commonly observed in DNA. This results in a very deep and
narrow major groove and a shallow and wide minor groove. A second
consequence of the presence of the 2'-hydroxyl group is that in conformationally
flexible regions of an RNA molecule (that is, not involved in formation of a double
helix), it can chemically attack the adjacent phosphodiester bond to cleave the
backbone.
Most cellular RNA molecules are single stranded. They may form secondary
structures such as stem-loop and hairpin.
Secondary structure of RNA. (a) stem-loop. (b) hairpin.
There are more unusual bases in RNA than in DNA. All normal RNA chains
either start with adenine or guanine: Three types of cellular RNA have been
distinguished:
Messenger RNA (mRNA) or template RNA
Ribosomal RNA (rRNA) and
Soluble RNA (sRNA) or transfer RNA (tRNA)
Ribosomal and transfer RNA comprise about 98% of all RNA. All three forms
of RNA are made on a DNA template.
Transfer RNA and messenger RNA are synthesized on DNA templates of the
chromosomes, while ribosomal RNA is derived from nucleolar DNA. The three types
of RNA are synthesized during different stages in early development. Most of the
RNA synthesized during cleavage is mRNA. Synthesis of tRNA occurs at the end or
cleavage, and rRNA synthesis begins during gastrulation.
Comparison between DNA and RNA
DNA RNA
1. DNA is the usual genetic material RNA is the genetic material of some
viruses.
2. DNA is usually double-stranded, Most cellular RNA is single stranded.
(In certain viruses DNA is single (Some viruses e.g. retrovirus, have
stranded, e.g. φ X 174). double stranded RNA).
3. The pentose sugar is deoxyribose. The pentose sugar is ribose.
4. The common organic bases are The common organic bases are
adenine, guanine, cytosine and adenine, guanine, cytosine and uracil.
thymine.
5. Base pairing: adenine pairs with Adenine pairs with uracil and guanine
thymine and guanine with with cytosine.
cytosine.
6. Pairing of bases is throughout the Pairing of bases is only in the helical
length of the molecule. region
7. There are fewer uncommon bases There are more uncommon bases.
8. DNA is only of one type There are three types of RNA
messenger, ribosomal and transfer
RNA.
9. Most of the DNA is found in the Messenger RNA is formed on the
chromosomes. Some DNA is also chromosomes, and is found in the
found in the cytoplasm e.g. in nucleolus and cytoplasm. rRNA and
mitochondria and chloroplasts. tRNA are also formed on the
chromosomes, and are found in
cytoplasm.
10. Denaturation (melting) is partially Complete and practically in
reversible only under certain stantaneous reversibility of the process
conditions of slow cooling of melting.
(renaturation).
11. Sharp, narrow temperature Broad temperature interval of
interval of transition in melting. transition in melting.
12. DNA on replication forms DNA, and Usually RNA does not replicate or
on transcription forms RNA. transcribe. (In certain viruses RNA can
synthesize an RNA chain).
13. Genetic messages are usually The usual function of RNA is
encoded in DNA. translating messages encoded in DNA
into proteins.
14. DNA consists of a large number of RNA consists of fewer nucleotides, up
nucleotides, up to 4.3 million to 12,000.
Ribosomal RNA – rRNA

Ribosomal RNA, as the name suggests, is found in the ribosomes. It
comprises about 80% of the total RNA of the cell. The base sequence of rRNA is
complementary to that of the region of DNA where it is synthesized.
In eukaryotes ribosomes are formed on the nucleolus. Ribosomal RNA is
formed from only a small section of the DNA molecule, and hence there is no
definite base relationship between rRNA and DNA as a whole.
Ribosomal RNA consists of a single strand twisted upon itself in some
regions. It has helical regions connected by intervening single strand regions. The
helical regions may show presence or absence of positive interaction. In the helical
region most of the base pairs are complementary, and are joined by hydrogen
bonds. In the unfolded single strand regions the bases have no complements.
Ribosomal RNA contains the four major RNA bases with a slight degree of
methylation, and shows differences in the relative proportions of the bases between
species. Its molecules appear to be single polynucleotide strands which are
unbranched and flexible. At low ionic strength rRNA behaves as a random coil, but
with increasing ionic strength the molecule shows helical regions produced by base
pairing between adenine and uracil and guanine and cytosine.
Hence rRNA does not show purine-pyrimidine equality. The rRNA strands
unfold upon heating and refold upon cooling. Ribosomal RNA is stable for at least
two generations. The ribosome consists of proteins and RNA. The 70S ribosome of
prokaryotes consists of a 30S subunit and a 50S subunit. The 30S subunit contains
16S rRNA, while the 50S subunit contains 23S and 5S rRNA.
The 80S eukaryote ribosome consists of a 40S and a 60S subunit. In
vertebrates the 40S subunit contains 18S rRNA, while the 60S subunit contains 28-
29S, 5.8S and 5S rRNA. In plants and invertebrates the 40S subunit contains 16-
18S RNA, while the 60S subunit contains 25S and 58 and 5.8S rRNA. There are
three types of ribosomal RNA on the basis of sedimentation and molecular weight.
Two of these classes are high molecular weight RNAs, while the third is a low
molecular weight RNA. The three classes are: (I) high molecular weight rRNA with
molecular weight of over a million, e.g. 21s-29s RNA, (2) high molecular weight
rRNA with molecular weight below a million e. g. 12-8-188 rRNA, (3) low molecular
weight rRNA e. g. 58 rRNA.
Messenger RNA - mRNA - Jacob and Monod (1961) proposed the name
messenger RNA for the RNA carrying information for protein synthesis from the
DNA (genes) to the sites of protein formation (ribosomes). It consists of only 3 to
5% of the total cellular RNA.
Size of Messenger RNA - mRNA - The molecular weight of an average sized
mRNA molecule is about 500,000, and its sedimentation coefficient is 8S. It should
be noted however, that mRNA varies greatly in length and molecular weight. Since
most proteins contain at least a hundred amino acid residues, mRNA must have at
least 100 X 3= 300 nucleotides on the basis of the triplet code.
Stability of Messenger RNA - mRNA - The cell does not contain large quantities
of mRNA. This is because mRNA, unlike other RNAs is constantly undergoing
breakdown. It is broken down to its constituent ribonucleotides by ribonucleases.
Structure of Messenger RNA - mRNA
Messenger RNA is always single stranded. It contains mostly the bases
adenine, guanine, cytosine and uracil. There are few unusual substituted bases.
Although there is a certain amount of random coiling in extracted mRNA, there is
no base pairing. In fact base pairing in the mRNA strand destroys its biological
activity
Since mRNA is transcribed on DNA (genes), its base sequence is
complementary to that of the segment of DNA on which it is transcribed. This has
been demonstrated by hybridization experiments in which artificial RNA, DNA
double strands are produced. Hydrization takes place only if the DNA and RNA
strands are complementary.
Usually each gene transcribes its own mRNA. Therefore, there are
approximately as many types of mRNA molecules as there are genes. There may be
1,000 to 10.000 different species of mRNA in a cell. These mRNA types differ only
in the sequence of their bases and in length.
When one gene (cistron) codes for a single mRNA strand the mRNA is said to
be monocistronic. In many cases, however, several adjacent cistrons may
transcribe an mRNA molecule, which is then said to be polycistronic or polygenic.
The mRNA molecule has the following structural features:
1. Cap. At the 5' end of the mRNA molecule in most eukaryote cells and animal
virus molecules is found a 'cap'. This is blocked methylated structure, m7Gpp Nmp
Np or m7Gpp Nmp Nmp Np. where: N = any of the four nucleotides and Nmp = 20
methyl ribose. The rate of protein synthesis depends upon the presence of the cap.
Without the cap mRNA molecules bind very poorly to the ribosomes.
2. Noncoding region 1 (NC1). The cap is followed by a region of 10 to 100
nucleotides. This region is rich in A and U residues, and does not translate protein.
3. The initiation codon is AUG in both prokaryotes and eukaryotes
4. The coding region consists of about 1,500 nucleotides on the average and
translates protein It is made up of 73-93 nucleotides (Rich and RajBhandary,
1976). Each bacterial cell probably contains about a hundred or more different
types of tRNA. The function of tRNA is to carry amino acids to mRNA during protein
synthesis. Each amino acid is carried by a specific tRNA. Since 20 amino acids are
coded to form proteins, it follows that there must be at least 20 types of tRNA.
It was formerly thought that only 20 tRNA molecular types exist, one for
each amino acid. It has, however, been shown that in several cases there are at
least two types of tRNA for each amino acid. Thus there are many more tRNA
molecules than amino acid types. These are probably coded by one gene.
Transfer RNA is synthesized in the nucleus on a DNA template. Only 0.025%
of DNA codes for tRNA. Synthesis of tRNA occurs near the end of cleavage stages.
Transfer RNA is an exception to other cellular RNAs in that a part of its
ribonucleotide sequence (-CCA) is added after it comes off the DNA template. Like
rRNA, tRNA is also formed from only a small section of the DNA molecule.
Therefore, it does not show any obvious base relationships to DNA. The tRNA
molecule consists of a single strand looped about it self. The 3' end always
terminates in a -C-C-A (cytosine- cytosine-adenine) sequence. The 5' end
terminates in G (guanine) or C (cytosine). Many of the bases are bonded to each
other, but there are also unpaired bases.
Transfer RNA - tRNA OR Soluble RNA – sRNA
After rRNA the second most common RNA in the cell is transfer RNA. It is
also called soluble RNA because it is too small to be precipitated by
ultracentrifugation at 100,000 g. It constitutes about 10-20% of the total RNA of
the cell. Transfer RNA is a relatively small RNA having a molecular weight of about
25,000 to 30,000 and the sedimentation coefficient of mature eukaryote tRNA is
3.8S.
Structure of Transfer RNA – tRNA
The nucleotide sequence (primary structure) of tRNA was first worked out by
Holley et al (1965) for yeast alanine tRNA. Since then the sequence of about 75
different tRNAs, ranging from bacteria to mammals, has been established. The
different tRNAs are all minor variants of the same basic type of structure.
Several models of the secondary structure of tRNA have been proposed, and
of these the cloverleaf model of Holley is the most widely accepted.
Transfer RNA (tRNA) is an essential component of the protein synthesis

reaction. There are at least twenty different kinds of tRNA in the cell1 and each one
serves as the carrier of a specific amino acid to the site of translation.
tRNA's are L-shaped molecules. The amino acid is attached to one end and
the other end consists of three anticodon nucleotides. The anticodon pairs with a
codon in messenger RNA (mRNA) ensuring that the correct amino acid is
incorporated into the growing polypeptide chain.
The L-shaped tRNA is formed from a small single-stranded RNA molecule that
folds into the proper conformation. Four different regions of double-stranded RNA
are formed during the folding process.
The two ends of the molecule form the acceptor stem region where the
amino acid is attached. The anticodon is an exposed single-stranded region in a
loop at the end of the anticodon arm.
The two other stem/loop structures are named after the modified nucleotides
that are found in those parts of the molecule. The D arm contains dihydrouridylate
residues while the TΨC arm contains a ribothymidylate residue (T), a
pseudouridylate residue (Ψ) and a cytidylate (C) residue in that order. All tRNA's
have a similar TΨC sequence. The variable arm is variable, just as you would
expect. In some tRNA's it is barely noticable while in others it is the largest arm.
tRNA's are usually drawn in the "cloverleaf" form (below) to emphasize the
base-pairs in the secondary structure.
Clover leaf model of tRNA
Unusual Bases in tRNA

In addition to the usual bases A, U, G and C, tRNA contain a number of
unusual bases, and in this respect differs from mRNA and rRNA. The unusual bases
of tRNA account for 15-20% of the total RNA of the cell. Most of the unusual bases
are formed by methylation (addition of -CHa or methyl group to the usual bases),
e.g. cytosine and guanine on methylation yield methylcytosine and methyl/guanine,
respectively.
Precursor tRNA molecules transcribed on the DNA template contains the usual
bases. These are then modified to unusual bases. The unusual bases are important
because they protect the tRNA molecule against degradation by RNase. This
protection is necessary because RNA is found floating freely in the cell.
Some of the unusual bases of tRNA are methyl guanine (GMe),
dimethylguanine(GMe2), methylcytosine (Me), ribothymine (T), pseudouridine (ψ),
dihydrouridine (DHU, H2U, UH2), inosine (I) and methylinosine (IMe, MeI). In
general, organisms high in the evolutionary scale contain more modified bases than
lower organisms.
Classification of tRNA - A Study of different tRNAs shows that the structure of the
acceptor stem, the anticodon arm and the TψC arm are constant. The differences in
the tRNAs lie in the D arm and the variable arm. Based on the differences in these
two variable regions, three classes of tRNA have been recognized.
Class I (D4-V4-5), with 4 base pairs in the D stem and 4-5 bases in the variable
loop.
Class II (DS-V4-5), with 3 base pairs in the D stem and 4-5 bases pairs in the
variable loop.
Class III (D3-VN), with 3 base pairs in the D stem and a large variable arm.
A simpler classification based only on the variable arm recognizes two types of
tRNA.
Class I with 4-5 bases in the variable loop
Class II with a large variable arm of 13-21 bases.
Tertiary Structure of Transfer - tRNA - Electron density maps have revealed
that tRNA has a tertiary structure. This structure is due to hydrogen bonds
(i) between bases,
(ii) between bases and ribose phosphate backbone and
(iii) between the backbone residues. (The hydrogen bonding in the double
helical stem regions of the tRNA molecular are considered to be in the
secondary structure).
Initiator Transfer RNA - tRNA
The starting amino acid in eukaryote protein synthesis is methionine, while in
prokaryotes it is N-formyl methionine. The tRNA molecule3 specific for these two
amino acids are methionyl tRNA (tRNAmet) and N-formyl- methionyl IRNA
(tRNAfmet) respectively.
These tRNAs are called initiator tRNAs, because they initiate protein
synthesis. Initiator tRNAs have certain features which distinguish them from other
tRNAs, and the initiator tRNAs of prokaryotes' and eukaryotes also differ.
In most prokaryotes the 5' terminal nucleoside is C. It has opposite it (i.e. in
the fifth position from the 3' end) an A nucleotide. There is no Watson-Crick base
pairing between the two. In the blue green 'alga' Anacystis nidulans, however, the
fifth nucleotide from the 3' end is C. In eukaryotes there is an A.U base pair at the
acceptor stem.
As noted previously, prokaryotes use tRNAf-met for initiation of protein

synthesis, while eukaryotes use tRNAmet. The prokaryote Halo bacterium
cutirubrum is, however, reported to initiate protein synthesis with tRNA met and
has an A.U base pair at the end of the accept or stem. In these respects it
resembles eukaryotes
The D loop of prokaryote initiator tRNAs contains an A11, U24 base pair. All
other tRNAs have a Y11, R24 base pair. Eukaryotic cytoplasmic initiator tRNAs have
AU or AU* instead of Tψ in the TψC loop. Also, in eukaryotes instead of a
pyrimidine nucleotide (Y) there is A at the 3' end of the TψC loop.
In some eukaryotic cytoplasmic initiator tRNAs the anticodon sequence CAU
is preceeded by C instead of U as in all other tRNAs. In prokaryotes the purine
nucleotide following C in the TψC loop is A, while in eukaryotes it is G. In tRNA f-
met the nucleotide adjacent to the 3' side of the anticodon triplet is adenosine while
in tRNA met it is alkylated adenosine
Specificity of Tranfer RNA - tRNA
Two important steps in translation during protein synthesis are the activation
of amino acids and the transfer of amino acids to tRNAs. Each amino acid has a
specific activating enzyme tRNA aminoacyl synthetase. Thus there are 20 different
tRNA aminoacyl synthetases for the 20 common amino acids found in proteins.
Some tRNA synthetases can activate more than one amino acid, i.e. they
show only a limited substrate specificity. Thus isoleucine tRNA synthetase can also
activate L valine, and valine tRNA synthetase can also react with threonine. The
enzymes, however, recognize only a specific set of, tRNAs as substratesL isolecine
tRNA synthetase recognizes only tRNAileu and valine tRNA synthetase recognizes
only tRNAval. Thus specificity is involved at two stages, activation of the amino acid
and transfer of the amino acid to tRNA. Another group of enzymes, the tRNA
aminoacyl transferases catalyse the transfer of an amino acid from the amino acid -
tRNA complex to specific acceptor molecules.
Lecture 17
TRANSCRIPTION – TRANSLATION - GENETIC CODE AND OUTLINE OF

PROTEIN SYNTHESIS
Central Dogma of Protein Synthesis

Proteins constitute the major part by dry weight of an actively growing cell.
They are widely distributed in living matter. All enzymes are proteins. Proteins are
built up from about 20 amino acids which constitute the basic building blocks. In
proteins the amino acids are linked up by peptide bonds to from long chains called
polypeptides.
The sequence of amino acids has a bearing on the properties of a protein,
and is characteristic for a particular protein. The basic mechanism of protein
synthesis is that DNA makes RNA, which in turn makes protein. The central dogma
of protein synthesis is expressed as follows:
DNA ----------------> DNA --------------------> RNA -------------> PROTEIN
Replication Transcription Translation
Noble Prize Winners in Protein Synthesis

Nobel prize winnes. In the last 15 years several Nobel prizes in physiology
and medicine have been awarded for work done on nucleic acids and protein
synthesis. In 1975 Alexander Todd of Great Britain was awarded the prize for his
studies on nucleotides and nucleotidic coenzymes.
The 1958 prize went to Beadle and Tatum for their work showing that one
gene is responsible for one enzyme. Lederberg also shared the prize for his work on
genetic recombination. The 1959 Nobel prize was shared by Ochoa and Kornberg
for successful synthesis of RNA and DNA, repectively.
Watson of U. S. A and Crick and Wilkins of Great Britain received the 1962
prize for elcucidating the structure of DNA. In 1965 the prize was awarded to Jacob,
Monod and Lwoff of France for their discovery of regulator genes.
The 1968 prize went to the Americans Nirenberg and Khorana for their work on the
genetic code, and to Holley (also of U. S. A.) for his finding out the nucleotide
sequence of tRNA. In 1969 Delbruck, Hershey and Luria received the prize for their
work on the reproductive pattern of viruses, In 1975 Temin was awarded the Nobel
prize for his work on RNA directed DNA synthesis.
PROTEIN SYNTHESIS
Proteins are widely used in cells to serve diverse functions. Some proteins
provide the structural support for cells while others act as enzymes to catalyze
certain reactions. We have already seen the roles that different enzymes play in
building the cell's structure and in catalyzing metabolic reactions, but where do
proteins come from?
Since the beginning of evolution, cells have developed the ability to
synthesize proteins. They can produce new proteins either for reproduction or to
simply replace a degraded one. To manufacture proteins, cells follow a very
systematic procedure that first transcribes DNA into mRNA and then translates the
mRNA into chains of amino acids. The amino acid chain then folds into specific
proteins.
Protein synthesis requires two steps: transcription and translation.
Ribonucleic acid (RNA) was discovered after DNA. DNA, with exceptions in
chloroplasts and mitochondria, is restricted to the nucleus (in eukaryotes, the
nucleoid region in prokaryotes). RNA occurs in the nucleus as well as in the
cytoplasm (also remember that it occurs as part of the ribosomes that line the
rough endoplasmic reticulum).
Crick's central dogma. Information flow (with the exception of reverse
transcription) is from DNA to RNA via the process of transcription, and thence to
protein via translation. Transcription is the making of an RNA molecule off a DNA
template. Translation is the construction of an amino acid sequence (polypeptide)
from an RNA molecule. Although originally called dogma, this idea has been tested
repeatedly with almost no exceptions to the rule being found.
Step 1: DNA Transcription

Protein synthesis begins in the cell's nucleus when the gene encoding a
protein is copied into RNA. Genes, in the form of DNA, are embedded in in the cell's
chromosomes. The process of transferring the gene's DNA into RNA is called
transcription. Transcription helps to magnify the amount of DNA by creating many
copies of RNA that can act as the template for protein synthesis. The RNA copy of
the gene is called the mRNA.
DNA and RNA are both constructed by a chain of nucleotides. However, RNA
differs from DNA by the substitution of uracil (U) for thymine (T). Also, because
only one strand of mRNA is needed when synthesizing proteins, mRNA naturally
exist in single-stranded forms.
After transcription, the mRNA is transported out of the cell's nucleus through
nuclear pores to go to the site of translation, the rough endoplasmic reticulum.
Transcription is a process of making an RNA strand from a DNA template,
and the RNA molecule that is made is called transcript. In the synthesis of proteins,
there are actually three types of RNA that participate and play different roles:
a. Messenger RNA(mRNA) which carries the genetic information from DNA and is
used as a template for protein synthesis.
b. Ribosomal RNA(rRNA) which is a major constituent of the cellular particles
called ribosomes on which protein synthesis actually takes place.
c. A set of transfer RNA(tRNA) molecules, each of which incorporates a
particular amino acid subunit into the growing protein when it recognizes a specific
group of three adjacent bases in the mRNA.
DNA maintains genetic information in the nucleus. RNA takes that
information into the cytoplasm, where the cell uses it to construct specific proteins,
RNA synthesis is transcription; protein synthesis is translation.
RNA differs from DNA in that it is single stranded, contains Uracil instead of
Thymine and ribose instead of deoxyribose, and has different functions. The central
dogma depicts RNA as a messenger between gene and protein, but does not
adequately describe RNA's other function.
Transcription is highly controlled and complex. In Prokaryotes, genes are
expressed as required, and in multicellular organisms, specialized cell types express
subsets of gene. Transcription factors recognize sequences near a gene and bind
sequentially, creating a binding transcription. Transcription proceeds as RNAP
inserts complementary RNA bases opposite the coding strand of DNA. Antisense
RNA blocks gene expression.
Messenger RNA transmits information in a gene to cellular structures that
build proteins. Each three mRNA bases in a row forms a codon that specifies a
particular amino acid. Ribosomal RNA and proteins form ribosomes, which
physically support the other participants in protein synthesis and help catalyze
formation of bonds betweens amino acids.
In eukaryotes, RNA is often altered before it is active. Messenger RNA gains
a cap of modified nucleotides and a poly A tail. Introns are transcribed and cut out,
and exons are reattached by ribozymes. RNA editing introduced bases changes that
alter the protein product in different cell types.
The genetic code is triplet, non-overlapping, continuous, universal, and
degenerate. As translation begins, mRNA, tRNA with bound amino acids, ribosomes,
energy molecules and protein factos assemble. The mRNA leader sequence binds to
rRNA in the small subunit of a ribosome, and the first codon attracts a tRNA bearing
methionine. Next, as the chain elongates, the large ribosomal subunit attaches and
the appropriate anticodon parts of tRNA molecules form peptide bonds, a
polypeptide grows. At a stop codon, protein synthesis ceases. Protein folding begins
as translation proceeds, with enzymes and chaperone proteins assisting the amino
acid chain in assuming its final functional form. Translation is efficient and
economical, as RNA, ribosomes, enzymes, and key proteins are recycled.
RNA transcription requires the following components
The enzyme RNA polymerase
 A DNA template
 All four types of ribonucleoside triphosphates (ATP, GTP and UTP)
 Divalent metal ions Mg++ or Mn++ as a co-factor
 No primer is needed for RNA synthesis
 RNA transcription is a process that involves the following steps.
Binding of RNA Polymerase to DNA Double Helix
The histone coat protecting the DNA double helix of the gene to be
transcribed is removed, on a signal from the cytoplasm, exposing the
polynucleotide sequences in this region of DNA. The RNA polymerase enzyme binds
to a specific site, called promoter, in the DNA double helix. This site is located on
the 5 side of the gene to be transcribed. It signals the beginning of RNA synthesis.
The promoter also determines the DNA strand that is to be transcribed.
Exposure of RNA Bases
The histone coat protecting the DNA double helix of the gene to be
transcribed is removed, on a signal from the cytoplasm, exposing the
polynucleotide sequences in this region of DNA. The RNA polymerase enzyme binds
to a specific site, called promoter, in the DNA double helix. This site is located on
the 5 side of the gene to be transcribed. It signals the beginning of RNA synthesis.
The promoter also determines the DNA strand that is to be transcription is not
known.
Base pairing
The ribonucleoside triphosphates, namely, adenosine triphosphate (ATP),
guanosine triphosphate (GTP), cytidine triphosphate (CTP) and uridine triphosphate
(UTP), floating free in the nucleus, serve as the raw material for RNA synthesis.
They are formed by activation (phosphorylation) of ribonucleoside
monophosphates, viz., adenosine monophosphate (AMP), guanosine
monophosphate (GMP), cytidine monophosphate (CMP) and uridine monophosphate
(UMP) as a result of their combining with ATP. The enzyme phosphorylase catalyses
this activation process. The ribonucleotide triphosphates are joined to the bases of
the DNA template chain one by one by hydrogen bonding according to the base
pairing rule i.e., A U, U A, C G, G C. This base pairing is brought about by the RNA
polymerase.
Synthesis of mRNA from DNA

The nucleotides are added one by one. A=Adenine, T=Thymine, C=Cytosine,
G=Guanine, U=Uracil, R=Ribose sugar, P=Phosphate
Conversion to Ribonucleoside Monophosphates
The various ribonucleoside triphosphates on linking to the DNA template

chain break off their high-energy bonds. This changes them to ribonucleoside
monophosphates which represent the normal components of RNA, and sets free
pyrophosphate groups (P~P). Pyrophosphate contains a high-energy bond (~). It
undergoes hydrolysis by the enzyme pyrophosphotase, releases energy and sets
free inorganic phosphate Pi. The first ribonucleotide phosphate retains all the three
phosphates and is, thus, chemically distinct from the other nucleotides added after
it
Formation of RNA Chain

Each ribonucleoside monophosphate attached to the DNA template chain
then combines with the ribonucleotide arrived earlier, making the RNA chain
become longer. The process is catalysed by the enzyme RNA polymerase and
requires a divalent ion Mg++ or Mn ++
. The RNA chain, thus formed, contains
nitrogenous bases that are complementary to those of the template DNA chain.
Separation of RNA Chain
As transcription proceeds, the hybrid DNA-RNA molecule dissociates, partly
releasing the RNA molecule under synthesis. When polymerase reaches a
terminator signal on the DNA, it leaves the DNA. The fully formed RNA chain is now
totally released by this process, one gene forms several molecules of RNA, which
get released from the DNA template one after the other.
In some cases, such as in E. coli, a specific chain terminating protein, called
rho factor (P), stops the synthesis of RNA chain. In most cases, the enzyme RNA
polymerase on its own can stop transcription.
Return of DNA Segment to Original Form
As the RNA chain grows, the transcribed region of the DNA molecule gets
hydrogen bonded to the opposite strand and the two become spirally coiled to
assume the original double helical form. When the last ribonucleotide is added, the
RNA polymerase and RNA chain are completely released from the DNA, and now the
DNA completes its winding into a double helix. The protective protein coat is added
again to the DNA duplex.
The sequence of nitrogen bases from the promoter to the terminator sites form a
transcription unit. It may include one or more genes. An entire transcription unit
gets transcribed into a single RNA chain.
Processing of RNAs
The forms of RNAs originally transcribed from DNA are called primary
transcripts. These undergo extensive changes, termed processing or post-
transcriptional modification of RNAs, before they can become functional in both
prokaryotes and eukaryotes.
In RNA processing,
 Larger RNA precursors are cut into smaller RNAs by a ribonuclease-P cleaving
enzyme
 Unwanted nucleotides are removed by enzymes called nucleases (splicing)
 Useful regions are rejoined by ligase enzyme
 Certain nucleotides are added at the terminal ends enzymatically (terminal
addition)
 The RNA molecule may fold on itself to assume proper shape (folding) and
 Some nucleotides may be modified (nucleotide modification)
The entire process of RNA transcription may be summed up in the equation
(ATP + GTP +CTP +UTP) n
RNA TYPES
The three different types of RNA, namely, messenger RNA (mRNA),
ribosomal RNA (rRNA) and transfer RNA (tRNA) are transcribed from different
regions of the DNA molecule. Three different RNA polymerases: I, II and III
catalyses the transcription of rRNA, mRNA and tRNA respectively in eukaryotes. In
prokaryotes, a single RNA polymerase composed of different subunits does this
work. Transcription of RNA also occurs in the 5-3 direction like the replication of
DNA.
Step 2: RNA Translation

After the mRNA has been transported to the rough endoplasmic reticulum, it
is fed into the ribosomal translation machineries. Ribosomes begins to read the
mRNA sequence from the 5` end to the 3` end. To convert the mRNA into protein,
tRNA is used to read the mRNA sequence, 3 nucleotides at a time.
Amino acids are represented by codons, which are 3-nucleotide RNA
sequences. The mRNA sequence is matched three nucleotides at a time to a
complementary set of three nucleotides in the anticodon region of the
corresponding tRNA molecule. Opposite the anticodon region of each tRNA, an
amino acid is attached and as the mRNA is read off, the amino acids on each tRNA
are joined together through peptide bonds.
Translation is the mechanism by which the triplet base sequence of an mRNA
guides the linking of a specific sequence of amino acids to form a polypeptide
(protein) on ribosomes. All the proteins a cell needs are synthesized by the cell
within itself.
Machinery for Protein Synthesis

Protein synthesis requires amino acids, DNA, RNAs, ribosomes and enzymes,
enzyme activators and ATP molecules
Amino Acids
Proteins are the polymers of amino acids. Therefore, amino acids form the
raw material for protein synthesis. The proteins found in living organisms need
about 20 amino acids as building blocks or monomers. These are available in the
cytoplasmic matrix as an amino acid pool.
DNA as Specificity Control
In order to maintain its own special characteristics a cell must manufacture
proteins exactly similar to those already present in it. Thus, protein synthesis
requires specificity control to provide instructions about the exact sequence in
which the given numbers and kinds of amino acids should be linked to form the
desired polypeptides. The specificity control is exercised by DNA through mRNA.
Sequences of 3 consecutive nitrogenous bases in the DNA double helix form the
biochemical or genetic code. Each base triplet codes for a specific amino acid. Since
the DNA is more or less stable, the proteins formed in a cell are exactly like the
preexisting proteins
RNAs
RNA molecule is a long, unbranched, single-stranded polymer of
ribonucleotides. Each nucleotide unit is composed of three smaller molecules, a
phosphate group, a 5-carbon ribose sugar, and a nitrogen-containing base. The
bases in RNA are adenine, guanine, uracil and cytosine. The various components
are linked up as in DNA. There are three types of RNA in every cell: messenger RNA
or mRNA, ribosomal RNA or rRNA and transfer RNA or tRNA. The three types of
RNAs are transcribed from different regions of DNA template. RNA chain is
complementary to the DNA strand, which produces it. All the three kinds of RNAs
play a role in protein synthesis.
Differences Between RNA Types
Ribosomal RNA Messenger RNA Transfer RNA

Features
(rRNA) (mRNA) (tRNA)
1. Percentage
of cell’s total About 80 About 5 About 15
RNA
2. Length of
Variable Longest Shortest
molecule
Clover leaf- like,

3.Shape of
Greatly coiled Linear folded into L-
molecule
shape
4. Types Six Numerous About 60
Form greater part Carry information from Carry amino acids

5. Role
of ribosome’s DNA to mRNA codons
Long, used again Very short, 2 minutes to Long, used again

6. Life and again in 4 hours, degraded after and again in
translation translation translation
mRNA
The DNA, that controls protein synthesis, is located in the chromosomes
within the nucleus, whereas the ribosomes, on which the protein synthesis actually
occurs, are placed in the cytoplasm. Therefore, some sort of agency must exist to
carry instructions from the DNA to the ribosomes. This agency does exist in the
form of mRNA. The mRNA molecule carries the message (information) from DNA
about the sequence of particular amino acids to be form a polypeptide, hence its
name. It is also called informational RNA or template RNA. The mRNA forms about
5% of the total RNA of a cell. Its molecule is linear and the longest of all the three
RNA types. Its length is related to the size of the polypeptide to be synthesized with
its information. There is a specific mRNA for each polypeptide. Because of the
variation is size in mRNA population in a cell; the mRNA is often called
heterogeneous nuclear RNA, or hnRNA
It has at its 5 end a cap of methylated guanine followed successively by an
initiation codon (AUG or GUG), a long coding region, a termination codon (UAA or
UAG or UGA) and a poly-A tail of many adenine-containing nucleotides at 3 end. A
small non-coding region may be present after the head and before tail
In eukaryotes, mRNA carries information for one polypeptide only. It is

monocistronic (monogenic) because it is transcribed from a single cistron (gene)
and has a single terminator codon
Bacterial mRNA often carries information for more than one polypeptide
chain. Such an mRNA is said to be polycistronic (polygenic) because it is
transcribed from many continuous (adjacent) genes. A polycistronic mRNA has an
initiator codon and a terminator codon for each polypeptide to be formed by it.
tRNA
The tRNA carries a specific amino acid from the amino acid pool to the mRNA on the
ribosomes to form a polypeptide, hence its name. The tRNAs form about 15% of
the total RNA of a cell. Its molecule is the smallest and has the form of a cloverleaf.
It has four regions.
Carrier End: This is the 3 end of the molecule. Here a specific amino acid becomes
attache d. The tRNA molecule has a base triplet CCA with OH group at the tip. The
COOH of amino acid joins the OH group.
Recognition End: It is the opposite end of the molecule. It has 3 unpaired
ribonucleotides. The bases of these ribonucleotides are complementary bases of the
triplet found on mRNA chain called a codon. This triplet base sequence in tRNA is
called as an anticodon. The anticodon binds with the codon at the time of
translation.
Enzyme Site: It is on one lateral side of the molecule. It is meant for a specific
charging enzyme which catalyses the binding of a specific amino acid to tRNA
molecule.
Ribosome Site: It is on the other lateral side of the molecule. It is meant for
attachment to a ribosome.
rRNA
The rRNA molecule is greatly coiled. In combination with proteins, it forms
the small and large subunits of the ribosomes, hence its name. It forms about 80%
of the total RNA of a cell. A eukaryotic ribosome is 80S; with a large 60S subunit
consists of 28S, 5.8S and 5S rRNAs and over 45 different basic proteins, the
smaller 40S subunit comprises 18S RNA and about 33 different basic proteins. A
prokaryotic ribosome is 70S; its large 50S subunit consists of 23S and 5S rRNAs
and about 34 different basic proteins; its small 30S subunit comprises 16S rRNA
and about 21 different basic proteins. The 3 end of 18S rRNA (16S rRNA in
prokaryotes) has a binding site for the mRNA cap. The 5S rRNA has a binding site
for tRNA. The rRNA also seems to play some general role in protein synthesis. It is
involved in assembling the amino acid molecules brought by tRNA, into a
polypeptide chain
There are two more types of RNA, recognised in the cell namely
Small nuclear RNA (snRNA) that helps in processing of rRNA and mRNA and
Small cytoplasmic RNA (scRNA) which helps in binding the ribosome to ER
Ribosomes
Ribosomes are tiny ribonucleoprotein particles without a covering membrane.

They serve as the site for protein synthesis. Hence, they are called protein factories
of the cell. Each ribosome consists of larger and smaller subunits. The subunits of
ribosome occur separately when ribosomes are not involved in protein synthesis.
The two subunits join when protein synthesis starts, and undergo dissociation
(separate) when protein synthesis stops. Many ribosomes line up on the mRNA
chain during protein synthesis. Such a group of active ribosomes is called a
polyribosome, or a polysome. In a polysome, the adjacent ribosomes are about 340
Ao apart. The number of ribosomes in a polysome is related to the length of the
mRNA molecule, which reflects the length of the polypeptides to be synthesized. It
is now known that polypeptide synthesis occurs at the polysomes and not at the
single free ribosomes, in both prokaryotes and eukaryotes.
A ribosome has two binding sites for tRNA molecules. One is called A site
(acceptor or aminoacyl) and the other is termed P site (peptidyl). These sites span
across the larger and smaller subunits of the ribosome. The A site receives the
tRNA amino acid complex. The tRNA leaves from P site, after releasing its amino
acid. However, the first tRNAamino acid complex directly enters the P site of the
ribosome.
A eukaryotic ribosome has a groove at the junction of the two subunits. From
this groove, a tunnel extends through the large subunit and opens into a canal of
the endoplasmic reticulum. The polypeptides are synthesized in the groove between
the two ribosomal subunits and pass through the tunnel of the large subunit into
the endoplasmic reticulum. While in the groove, the developing polypeptide is
protected from the cellular enzymes.
The smaller subunit forms a cap over the larger subunit. The larger subunit
attaches to the endoplasmic reticulum by two glycoproteins named ribophorin I and
II.
The function of the ribosome is to hold the mRNA, tRNA and the associated
enzymes controlling the process in position, until a peptide bond is formed between
the adjacent amino acids
Mechanism of Protein Synthesis
The events in protein synthesis are better known in bacteria than in
eukaryotes. Although these are thought to be similar in the two groups there are
some differences. The following description refers mainly to protein synthesis in
bacteria on the 70S ribosome.
Protein synthesis is a highly complex and an elaborate process and involves the
following steps:
Activation of Amino Acids
It is the step in which each of the participating amino acid reacts with ATP to
form amino acid AMP complex and pyrophosphate. The reaction is catalyzed by a
specific amino acid activating enzyme called aminoacyl-tRNA synthetase in the
presence of Mg2+. There is a separate aminoacyl tRNA synthetase enzyme for each
kind of amino acid. Much of the energy released by the separation of phosphate
groups from ATP is trapped in the amino acid AMP complex. The complex remains
temporarily associated with the enzyme. The amino acid AMP enzyme complex is
called an activated amino acid. The pyrophosphate is hydrolyzed to two in organic
phosphates (2pi)
2+
Activatiing enzyme Mg
Amino acid + ATPAmino acid AMP enzyme complex + ppi
Activation of Amino Acids

Charging of tRNA
It is the step in which the amino acid AMP-enzyme complex joins with the
amino acid binding site of its specific tRNA, where its COOH group bonds with the
OH group of the terminal base triplet CCA. The reaction is catalyzed by the same
enzyme, aminoacyl tRNA synthetase. The resulting tRNA-amino acid complex is
called a charged tRNA. AMP and enzyme are released. The released enzyme can
activate and attach another amino acid molecule to another tRNA molecule. The
energy released by change of ATP to AMP is retained in the amino acid-tRNA
complex. This energy is later used to drive the formation of peptide bond when
amino acids link together and form a polypeptide
Amino acid AMP Enzyme complex +t RNA
The tRNA amino acid complex moves to the ribosomes, the site of protein
synthesis.
Activation of Ribosome
It is the step in which the smaller and the larger subunits of ribosome are
joined together. This is brought about by mRNA chain. The latter joins the smaller
ribosomal subunit with the help of the first codon by a base pairing with an
appropriate sequence on rRNA. The combination of the two is called initiation
complex. The larger subunit later joins the small subunit, forming active ribosome.
++
Activation of ribosome by mRNA requires proper concentration of Mg
Assembly of Amino Acids (Polypeptide Formation)
It is the step in which the amino acids are assembled into a polypeptide
chain. It involves 3 events: initiation, elongation and termination of polypeptide
chain
Initiation of Polypeptide Chain
The mRNA chain has at its 5 end an “initiator” or “start” codon (AUG or GUG)
that signals the beginning of polypeptide formation. This codon lies close to the P
site of the ribosome. The amino acid formylmethionine (methionine in eukaryotes)
initiates the process. It is carried by tRNA having an anticodon UAC which bonds
with the initiator codon AUG of mRNA. Initiation factors (IF1, IF2 and IF3) and GTP
promote the initiation process.
The large ribosomal subunit now joins the small subunit to complete the
ribosome. At this stage, GTP is hydrolysed to GDP. The ribosome has
formylmethionine bearing tRNA at the P site. Later, the formylmethionine is
changed to normal methionine by the enzyme deformylase in prokaryotes. If not
required, methionine is later separated from the polypeptide chain by a proteolytic
enzyme aminopeptidase.
Elongation of Polypeptide Chain
The above figure shows,

A. A charged tRNA arriving at the A site, reading its codon on the mRNA
B. Amino acid of tRNA at P site is ready to be transferred to the amino acid of tRNA
at A site
C. Amino acids are joined by peptide bond and tRNA is discharged from P site
D. Peptide chain-carrying tRNA is translocated to P site, making A site free to
receive another charged tRNA
Three elongation factors (EF Tu, EF Ts and EF G) assist in the elongation of
the polypeptide chain. A charged tRNA molecule along with its amino acid, proline,
for example, enters the ribosome at the A site. Its anticodon GGA locates and binds
with the complementary codon CCU of mRNA chain by hydrogen bonds. The amino
acid methionine is transferred from its tRNA onto the newly arrived proline tRNA
complex where the two amino acids join by a peptide bond. The process is
catalyzed by the enzyme peptidyl transferase located on the ribosome. In this
process, the linkage between the first amino acid and its tRNA is broken, and the -
COOH group now forms a peptide bond with the free -NH2 group of the second
amino acid. Thus, the second tRNA carries a dipeptide, formylmethionineproline.
The energy required for the formation of a peptide bond comes from the free
energy released by separation of amino acid (formylmethionine or methionine) from
its tRNA.
The first tRNA, now uncharged, separates from mRNA chain at the P site of
the ribosome and returns to the mixed pool of tRNAs in the cytoplasm. Here, it is
now available to transport another molecule of its specific amino acid.
Now the ribosome moves one codon along the mRNA in the 3 direction. With
this, tRNAdipeptide complex at the A site is pulled to the P site. This process is
called translocation. It requires GTP and a translocase protein called EF-G factor.
The GTP is hydrolysed to GDP and inorganic phosphate to release energy for the
process
At this stage, a third tRNA molecule with its own specific amino acid,
arginine, for example arrives at the A site of the ribosome and binds with the help
of anticodon AGA to the complementary codon UCU of the mRNA chain. The
dipeptide formylmethionineproline is shifted from the preceding tRNA on the third
tRNA where it joins the amino acid arginine again with the help of peptidyl
transferase enzyme. The dipeptide, thus, becomes a tripeptide, formyl-methionine-
proline-arginine. The second tRNA being now uncharged, leaves the mRNA chain,
vacating the P site. The tRNAtripeptide complex is translocated from A site to P site
The entire process involving arrival of tRNA-amino acid complex, peptide bond
formation and translocation is repeated. As the ribosome moves over the mRNA, all
the codons of mRNA arrive at the A site one after another, and the peptide chain
grows. Thus, the amino acids are linked up into a polypeptide in a sequence
communicated by the DNA through the mRNA. A polypeptide chain which is in the
process of synthesis is often called a nascent polypeptide
The growing polypeptide chain always remains attached to its original
ribosome, and is not transferred from one ribosome to another. Only one
polypeptide chain can be synthesized at a time on a given ribosome.
Termination and Release of Polypeptide Chain:
At the terminal end of mRNA chain there is a stop, or terminator codon (UAA,
UAG or UGA). It is not joined by the anticodon of any tRNA amino acid complex.
Hence, there can be no further addition of amino acids to the polypeptide chain.
The linkage between the last tRNA and the polypeptide chain is broken by three
release factors. (RF 1, RF 2 and RF 3) and GTP. The release is catalyzed by the
peptidyl transferase enzyme, the same enzyme that forms the peptide bonds. The
ribosome jumps off the mRNA chain at the stop codon and dissociates into its two
subunits. The completed polypeptide (amino acid chain) becomes free in the
cytoplasm.
The ribosomes and the tRNAs on release from the mRNA can function again
in the same manner and result in the formation of another polypeptide of the same
protein.
Modification of Released Polypeptide
The just released polypeptide is a straight, linear exhibiting a primary
molecule, structure. It may lose some amino acids from the end with the help of a
peptidase enzyme, and then coil and fold on itself to acquire secondary and tertiary
structure. It may even combine with other polypeptides, to have quaternary
structure.
The proteins synthesized on free polysomes are released into the cytoplasm
and function as structural and enzymatic proteins. The proteins formed on the
polysomes attached to ER pass into the ER channels and are exported as cell
secretions by exocytosis after packaging in the Golgi apparatus.
Polysome Formation
When the ribosome has moved sufficiently down the mRNA chain towards 3
end, another ribosome takes up position at the initiator codon of mRNA, and starts
synthesis of a second molecule of the same polypeptide chain. At any given time,
the mRNA chain will, therefore, carry many ribosomes over which are similar
polypeptide chains of varying length, shortest near the initiator codon and longest
near the terminator codon. A row of ribosomes joined to the mRNA molecule, is
called a polyribosome, or a polysome. Synthesis of many molecules of the same
polypeptide simultaneously from one mRNA molecule by a polysome is called
translational amplification.
Energy Used for Protein Synthesis

One GTP is hydrolysed to GDP as each successive amino acid-tRNA complex
attaches to the A site of the ribosome. A second GTP is broken down to GDP as the
ribosome moves to each new codon in the mRNA. One ATP is hydrolysed to AMP
during amino acid activation. Thus, the formation of each peptide bond uses 3 high-
energy molecules, one ATP and two GTP.
An interesting aspect of protein synthesis is that the DNA and ribosomes are
located at different sites in the cell. Location of instruction centre (DNA) and
manufacturing centre (ribosomes) at different sites in a cell is advantageous. If
both were in the nucleus, the manufacturing centre would be far away from the
energy sources and raw materials; and if both were in the cytoplasm, the
information centre would be exposed to respiratory breakdown. The nuclear
envelope preserves stability of the DNA by protecting it from respiratory

destruction. The message in the DNA in the form of genes (codes) are, permanent,
authentic master documents from which working copies are prepared in the form of
mRNAs, as and when required by the cell.
The complex process by which the information in RNA is decoded into a
polypeptide is one of the exciting discoveries the genetic code
With only four biochemical letters (A, G, C, U) a one letter code could not
unambiguously encode 20 amino acids. A two letter code could encode only 16
amino acids. So, a triplet code based on 3 biochemical letters or nucleotide bases
could make 4 x 4 x 4 = 64 codons. This will be required to code for 20 or so
different amino acids.
The discovery of the genetic code became possible through the contribution
of many scientists like Francis Crick, Seveno Ochoa, Maxell Ninenberg, Hargobind
Khorana and J. H. Malther in the 1960s. Ninemberg and Khorana shared the Nobel
Prize in 1968.
Characteristics of Genetic Code
The genetic code is a triplet code: Three adjacent bases, termed as codon,
specify one amino acid
Non-overlapping: Adjacent codons do not overlap
No punctuation: The genetic code is comma less
The genetic code is universal i.e., a given codon specifies the same amino acid in all
protein synthesising organisms
The genetic code is degenerate: it lacks specificity and one amino acid often has
more than one code triplet
Each codon codes for only one amino acid, none for more than one
Three of the 64 codons, names UAA, UAG and UGA do not specify any amino acid
but signal the end of the message. They are called nonsense or terminator codons
The codons AUG and GUG are called the initiation or start codons as they begin the
synthesis of polypeptide.
In the above figure, the sequence of nucleotides in the triplet codons of RNA is
indicated; each triplet specifies a particular amino acid.
Lecture 18
CROSSING OVER - FACTORS AFFECTING IT

Crossing over
Genetic recombination crossing over is the mutual exchange of the
corresponding parts of the adjacent paternal and maternal chromatids of the
pachytene of meiosis I, producing new combinations of genes. The chromatids
resulting from the interchange of segments are known as the cross over
recombinants and the chromatids that remain intact are called non-crossover
parental chromatids.
A :Single Crossing, B :Reciprocal Double Crossing Over, C and D: Complementary Double

Crossing Over
It produces 50% parental and 50% recombinant chromatids

Significance of Crossing Over
a. It produces new individuals having new combinations of traits.
b. Crossing over has helped in establishing the concept of linear arrangement of
genes.
c. The frequency of crossing over helps in the mapping of chromosomes. i.e.,
determining the location of the genes in the chromosomes.
d. Selection of useful recombination by geneticists has brought about green
revolution in our country.
Stern’s Drosophila Experiments
In 1931, a few weeks after publication of similar experiments by Creighton
and McClintock in Zea mays Stern reported work with Drosophila.
There are 2 Linked traits: carnation eye and bar-eye flies.Carnation is recessive,
bar-eye is dominant. In B/B homozygotes, the eye is very narrow. In B/+ heteros,
get “wide-bar”.
Male had normal X chromosome, car +/Y and Female had 2 cytologically
distinct X chromosomes. One X had a portion of the Y chromosome attached to it,
making it longer than normal. This one was wild type for both genes.The other X
was shorter than normal, and had the car and B alleles on it.
The observed results require the above types of gametes to be produced by

females. The key observation was that in every case of genetic recombination,
there had also been recombination of the cytological features . The same type of
results had been found in the corn experiments. Genetic recombination
(phenotypes of the offspring) was always accompanied by recombination of the
cytological features. Together, these two studies convinced the field of genetics that
physical exchange of chromosome segments was the basis for recombination of

linked genes.
BUT, when did the crossing-over take place. Remember, at the start of
meiosis, the homologues are already replicated, and already paired up. So, one
plausible model is that crossing-over actually takes place during interphase, before
replication. That is, when each chromosome pair would consist of 2 chromatids, not
4 (not the tetrad stage).
How to distinguish between the two models?

To distinguish between the two models a right model organism
Neurospora crassa (a bread mold) was selected.
Normally, it reproduces asexually and also sexually. There are two mating
types: A and a . The 2 haploid nuclei fuse to give a diploid nucleus, the only diploid
phase to this life cycle. Meiosis produces 4 haploid spores within an ascus.
Subsequent mitotic division produces 8 haploid, sexual ascospores in a linear
arrangement. All the products of a single meiosis are found together in the ascus.
and, the order of the 4 spore pairs is the same as the order of the 4 chromatids of
each tetrad during Metaphase I. By capitalizing on the biology of our model
organism, we can now rigorously test our 2 competing hypotheses:Does crossing
over take place during the 2-chromatid phase, or during the 4-chromatid phase?
To test, we need strains with mutations at 2 genes on the same

chromosome. In Neurospora, there are many nutritional mutants are known. These
are unable to make some essential nutrient on their own, and it must be supplied in
their medium. met mutants are unable to synthesize their own methionine. his
mutants are unable to synthesize their own histidine.
All spores are recombinants.
Note: 1/2 are parental, 1/2 are recombinant. Repeated experiments like this
have shown that we get the second type of result. I.e., crossing over takes place at
the tetrad stage (4 chromatids). They also show that the exchange is reciprocal:
equal amounts of the 2 chromosomes are swapped.
CHROMOSOME MAP
( Linkage map / Cross over map / Genetic map)
The chromosome map may be defined as a line, on which the genes are
represented by points, separated by distances proportional to the amount of
crossing over.
The chromosome maps are also referred to as cross over maps since they
are sketched by the amount of crossing over.
The percentage of crossing over is directly proportional to the distance of the

alleles showing crossing over in the chromosome.
The chromosomes maps are the graphic representation of the genes in a

chromosome.
The percentage of crossing over is calculated by test crosses. In mapping the

genes, a unit of distance is used and it is called as map unit or Morgan unit.
The first chromosome map was made in 1911 by Sturtevant and soon after
additional maps were made by Bridges and others.
Drosophila is the earliest material used by the scientists, for constructing

maps.
Procedure for the chromosome mapping

In fact genes are plotted on the chromosome on the basis of crossing over
results between different pairs of linked genes. The actual distance between two
genes is said to be equivalent to the percentage of crossing over between these
genes.
When the % of crossing over between two genes is 5, then the distance is 5
units. For example five genes A, B, C, D and E are to be plotted on a chromosome.
If cross over results indicate that genes A and E have the highest percentage of
crossing over, it means that these should be placed at the maximum distance.
In this example, the gene A can be taken as a starting point in the
chromosome and can be represented by O.
Now if the gene A and B exhibit 7% crossing over, the gene B can be placed
on the chromosome at a distance of 7 units.
If the gene C shows 8% crossing over with gene B and about 15% crossing
over with gene A, it can be plotted on the chromosome at a distance of 15 units
from gene A.
Similarly if gene A and E exhibit 20% and 30% crossing over with gene D
and 5% and 10% with gene C these, are located on the chromosome 5 and 10
units away from the gene C respectively.
Construction of Chromosome map in Drosophila
In Drosophila the chromosome map is constructed with the help of test cross.
In Drosophila grey colour is dominant over black colour; and the long wing is
dominant over vestigial wing.
The F 1 female hybrid is test crossed. Four types of individuals are formed.
Out of four types, two types are parental type (G:L & B:V) and other two are non
parental type (G:V & B:L) due to crossing over. Non – parental type is 17%. So the
percentage of crossing over is equivalents to 17%. The distance between the two
genes (G-L) is equivalent to the percentage of crossing over or percentage of non
parental combination. So the distance between the gene G & L is equivalent to 17
morgan units.
Percentage of non parental combination = 17%
So the percentage of crossing over = 17
So the distance between the Gene G & L = 17 map unit
In another experiment the F 1 female grey red is test crossed with black
cinnabar. The experiment shows 9% non parental combination individuals. So the
distance between the Gene G & Cn is equavalent to 9 map unit.
In the same way the F 1 female red long is test crossed with cinnabar
vestigial. The experiment shows 9.5% non-parental combination individuals. So the
distance between the gene Cn is equivalent to 9.5 map unit.
According to the first experiment the distance between G & L is equivalent to

17 map unit. But the second and third experiment show 18.5 map units between
the two genes. To find out the actual reason for this difference in the distance,
conduct a 3 point cross.
Three Point cross
In the three point cross all the three pairs of genes are considered in the
experiment. The F 1 hybrid female is test crossed. They produce 8 different types of
individuals. Out of 8 types, two types are parental. Remaining six are non-parental.
Male Female
Parent : Grey Red Long Black cinnabar Vestigial
G Cn L x g cn l
Y-chromosome g cn l
Normal
F1 : G Cn L
g cn l
Back cross : Female Male
Normal Recessive
G Cn L x g cn l
g cn l Y – chromosome
F 2 Offspring
Parental combinations due to linkage = 658 = 82.25%
Non parental combination : Single cross over :
a) Between G and Cn g Cn L = 36
1. Black red long g cn l = 70
2. Grey cinnabar vestigial G cn l = 34
g cn l
b) Between Cn and L
1. Grey red vestigial G Cn l
g cn l = 35
= 66
2. Black cinnabar long g cn L = 31
g cn l
Double cross over
1. Grey cinnabar long G cn L = 4
g cn l = 6
2. Black red vestigial g Cn l = 2
g cn l
Total c.o between G and Cn = 70 + 6 = 76 = 9.5%
(includes double c.o)
Total c.o between Cn and L = 66 + 6 = 72 = 9.0%
Total c.o. between G and L = 76 + 72 = 148 = 18.5%
C.o between G and E = 70 + 66 = 136 = 17%
(double c.o not included)
From these results, it is concluded that the gene cinnabar lies about half –
way between the genes for black body colour and vestigial wings. The total amount
of crossing over between black body and vestigial wing is 18.5% rather than the
17% expected on the basis of the first cross. The discrepancy (18.5 – 17 = 1.5)
just noted, arises because of the occurrence of double crossing over, that is, of two
cross overs occurring simultaneously in the same cell between these two loci.
As a final check on these results, it would be well to make a trihybrid or three

point cross using all three pairs of genes at once. When pure recessive flies are
crossed with normal flies, all the F 1 flies are normal phenotypically. When the F 1
females are back crossed to triple recessive males, eight phenotypes are obtained.
From the data obtained the relative position of the genes can be calculated.
The distance between G and Cn is 9 units, the distance between Cn and L is 9.5
units; the L gene could be to the right of Gn locus or to the left. If the first order
(CnL) is correct, then the distance between G and L is 17 units. This small
discrepancy is due to double crossing over. Based upon the above data, the three
genes can be mapped as follows :
Chromosome Maps of Drosophila

The chromosome maps of Drosophila include four linkage groups
corresponding to four chromosome pairs. The genes present in the X chromosome
constitute the first linkage group, those present in 2nd and 3rd chromosome
constitute 2nd and 3rd linkage groups and those on the fourth chromosome form
fourth linkage group. The fourth linkage group is the smallest of all.
Chromosome maps of maize
Chromosome maps of maize have been drawn by R.A. Emerson. As there are
10 pairs of chromosome 10 chromosome maps are seen.
Factors affecting the mapping

Chromosome map can be constructed only with the help of crossing over
percentage. The crossing over percentage is highly modified by the interference and
coincidence.
Interference
Normally the double crossing over frequency is very low. Because the
crossing over and chiasma formation in the homologous non sister chromatids
interferes with the crossing over and chiasma formation at other points nearby.
This is called as interference. This was discovered by Muller (1911). The
interference is inversely proportional to the crossing over percentage. The
interference is maximum over a short distance and decreases as the distance

increases.
Coincidence
The coincidence is an inverse measure of interference. It is measured as a
ratio between actual number of double cross overs and the expected number of
double cross overs.
Actual number of double cross overs
Coindicidence = Expected number of double cross overs
If the actual number of double cross overs is zero, then coincidence is zero
and interference is complete. If the actual number of double cross overs is the
same as the expected number, coincidence is said to be one, and interference is nil.
It ranges from 0 to 1.
Lecture 19
LINKAGE - TYPES OF LINKAGE AND ESTIMATION OF LINKAGE.

LINKAGE
Every individual organism bears several heritable characters which are
represented by the innumerable genes present on the chromosomes. During
meiosis, the chromosomes move into the gametes as units, all the genes present
on any given chromosome will segregate as a group and move together from
generation to generation. This tendency of the genes located on the same
chromosome, to stay together in hereditary transmission, is known as linkage. The
genes located on the same chromosome are called linked genes.
The principle of linkage was discovered by Bateson and Punnet in 1906 in the
sweat pea, plant, Lathyrus odoratus. However, linkage, as a concept was put forth
by Thomas Hunt Morgan in 1910 based on his experiment on Drosophila
melanogaster.
Chromosome Theory of Linkage
Morgan, along with Castle formulated the chromosome theory of linkage. It has the
following postulates;
1. Genes are found arranged in a linear manner in the chromosomes.
2. Genes which exhibit linkage are located on the same chromosome.
3. Genes generally tend to stay in parental combination, except in cases of crossing
over.
4. The distance between linked genes in a chromosome determines the strength of
linkage. Genes located close to each other show stronger linkage than that are
located far from each other, since the former are less likely to enter into crossing
over.
Linkage Groups
All the genes located on a particular chromosome, form a linkage group.
Since, the genes present on a particular chromosome have their alleles located on
its homologous chromosome, genes on a pair of homologous chromosomes. Hence,
the number of linkage groups corresponds to the number of haploid chromosomes
found in a species.
Drosophila melanogaster has four linkage groups which can be distinguished
into three large and one small linkage groups corresponding to the four pairs of
chromosomes. Twenty-three linkage groups are present in humans corresponding

to 23 pairs of chromosomes.
Pea plant has seven linkage groups, corresponding to the seven pairs of
chromosomes.
Linkage in maize
'C' for coloured aleurone is dominant over 'C' colourless
Sh for Full endosperm is dominant over 'sh' shrunken.
F2 did not show 9: 3: 3: 1 ratio. There were greater number of colour full,
colour shrunken (parental types) than colourfull shrunkern , colour full, If two
character considered separately,they segregate 3 : 1
i.e . Colour - 7500 Full - 7500
Colouless - 2500 Shrunken - 2500
The large deviation of the observed F2 population form the excepted

segregation is therefore not because the members of each pair of alleles do not
segregate from each other but because of the separation in one pair of alleles is not
independent of the separation in the other pair of alleles.
Segregation of two pairs of genes on two pairs of chromosomes

Let us suppose that, gene 'C' is located on chromosome number 9 and 'S' on
chromosome number 10 of maize. The segregation of chromosome bearing C and c
is entirely independent of segregation of chromosome bearing S and s. So four type
of gametes Cs, Cs, eS, eS are formed in F1 and F2 normal dihybrid ratio 9:3:3:1
and test cross 1:1:1:1
Segregation for two pairs of genes on one pair of chromosomes
Let us suppose that, two genes C and S are located on chromosome No. 9
during meiosis only 2 gametes will be formed Cs and cs gametes.So, Genes C and S
situated on same chromosomes are said to be linked. Linkage is the association of
character in inheritance due to fact that genes determining them are physically
located on the same chromosomes.
Detection of Linkage
Compare the number of individuals observed in each class with those
expected on the basis of independent assortment and then to test the deviation
between these two values by chi-square test.
Linkage Group
The number of linkage groups will be equal to the haploid number of

chromosomes which the species possess. Thus maize has 10 pairs chromosomes
has 10 linkage groups.
Symbol of linked genes
While representing linked gene, the two homologous chromosomes are indicated
by two horizontal links.
Coupling
In the condition is linked inheritance in which an individual heterozygous for
two pairs of genes receives the two dominant member from one parent and the two
recessive members from the other parent.
Repulsion is the condition is linked inheritance, in which an individual

heterozygous for two pairs of linked genes receives the dominant member of one
pair and the recessive member of the other pair from one parent and the reverse
from the other parent
Crossing over
Leading to recombination of linked genes is due to the exchange of
corresponding segments between the chromatids of homologous chromosomes and
was first observed by Belgian cytologist Janssens in 1909.
Linkage studies revealed the following

1. Genes that assort at random are non linked genes. Genes that do not segregate
at random are linked genes.
2. Linked genes are arranged in a lines fashion on the chromosome. Each linked
gene has a definite and constant order in its arrangement.
3. The distance between the linked genes determines the degree of strength of
linkage. Closely located genes show stronger linkage that the widely located genes.
4. Linked genes do not always stay together, but are often exchanged reciprocally
by cross over.
Complete Linkage
The genes closely located in the chromosome show complete linkage as they
have no chance of separating by crossing over and are always transmitted together
to the same gamete and the same offspring. Thus, the parental combination of
traits is inherited as such by the young one.
Incomplete Linkage
The genes distantly located in the chromosome show incomplete linkage
because they have a chance of separation by crossing over and of going into
different gametes and offspring.
Importance of linkage in breeding
When there is a close linkage between desirable and undesirable characters
these genes are inherited in blocks and not individually and recombination is
practically nil. In such cases linkage has to be broken by ' irradiation'.
Linked Genes on the Same Chromosome Exhibit Distorted Mendelian Ratios
It was not long from the time that Mendel's work was rediscovered that new
anomalous ratio began appearing. One such experiment was performed by Bateson
and Punnet with sweet peas. They performed a typical dihybrid cross between one
pure line with purple flowers and long pollen grains and a second pure line with red
flowers and round pollen grains. Because they knew that purple flowers and long
pollen grains were both dominant, they expected a typical 9:3:3:1 ratio when the
F 1 plants were crossed. The table below shows the ratios that they observed.
Specifically, the two parental classes, purple, long and red, round, were over
represented in the progeny.
Observed Expected
Purple, long
284 215
(P_L_)
Purple, round
21 71
(P_ll)
Red, long
21 71
(ppL_)
Red, round
55 24
(ppll)
Total 381 381
At the time of these experiments, Bateson and Punnett were not able to
develop an acceptable hypothesis. The best explanation they posed was that in
some manner the phenotypic classes (alleles) in the parents were coupled, and
they did not sort independently into gametes as predicted by Mendel's second law.
Proof those genes on the same chromosome can at times be inherited as
blocks awaited the results of Thomas Hunt Morgan with Drosophila. Morgan crossed
red eye, normal wing flies (pr+pr+ vg+vg+) with purple eye, vestigal wing (prpr
vgvg) flies. The figure below shows the cross and the F 1 genotypes. (The bars are
used to shows that the genes reside on the same chromosome.)During meiosis,
four different F 1 gametes are produced. The parental gametes are developed
without any processing. The recombinant gametes though occur by a process
called crossing over. (The X between the two F 1 chromosomes represents the
crossing over event.)
Morgan performed a testcross by crossing prpr vgvg flies to F 1 . The testcross

is powerful because it allows you to follow the meiotic events in one parent because
all of the gametes from the test cross parent are homozygous recessive. For this
example, the testcross genotype is pr vg. Therefore the testcross progeny will
represent the distribution of the gametes in the F 1 . Remember that a testcross to
F 1 derived from a dihybrid cross gave a 1:1:1:1 ratio. But this is not what Morgan
observed. The following table shows the result of this test cross.
F 1 Gamete Testcross Distribution Gamete Type
pr+ vg+ 1339 Parental
pr+ vg 151 Recombinant
pr vg+ 154 Recombinant
pr vg 1195 Parental
These results confirm the Bateson and Punnett hypothesis that two genes do
not always assort independently. A further confirmation experiment was performed
by Morgan when he crossed red eye, vestigal wing flies and purple eye, normal
wing flies. Whereas in the first cross, the two dominant alleles and two recessive
alleles were on the same chromosome the F 1 , in the is cross a dominant allele was
on the same chromosome as a recessive allele. The term for the first chromosomal
arrangement of the F 1 is called coupling, whereas the second arrangement is
called repulsion. Another set of terms to describe these arrangements are cis and
trans, respectively. The following shows the chromosomal arrangement for the
cross of two parents in repulsion.
As with the first cross, Morgan test crossed these F 1 flies. The following table shows
the distribution of these F 1 gametes.
F 1 Gamete Testcross Distribution Gamete Type
pr+ vg+ 157 Recombinant
pr+ vg 965 Parental
pr vg+ 1067 Parental
pr vg 146 Recombinant
It was expected that both the coupling and repulsion crosses would yield
1:1:1:1 ratios. How can we determine if the results deviate from this ratio. As with
any ratio, we can use the chi-square test to determine if the observed results fit or
deviate from the expected ratio. The two tables below show the results for the chi-
square for the two crosses.
Coupling Cross Chi-Square Test
F 1 Gamete Observed Expected (O-E)2/E
pr+ vg+ 1339 709.75 557.9
pr+ vg 151 709.75 439.9
pr vg+ 154 709.75 435.2
pr vg 1195 709.75 331.8
Total 2839 2839 X2=1764.8
Repulsion Cross Chi-Square Test
F 1 Gamete Observed Expected (O-E)2/E
pr+ vg+ 157 583.75 312.0
pr+ vg 965 583.75 249.0
pr vg+ 1067 583.75 483.3
pr vg 146 583.75 328.3
Total 2335 2335 X2=1372.6
It is quite clear that both of these large chi-square values indicate that neither of
these ratios fit the 1:1:1:1
Lecture 20
NUMERICAL MUTATIONS (POLYPLOIDY)
Mutations which alter the chromosome structure, size or gene arrangement

are chromosomal mutations. Chromosomal mutations are widely called as
chromosomal aberrations. These are grouped into two broad classes based open
whether they alter the structure or number of chromosomes.
Chromosome Mutations - gross changes in chromosomes.
Changes in the number of chromosomes.
1. Euploidy - variation in the number of sets of chromosomes.
a. Haploidy (Monoploidy) - one set of chromosomes (n) : ABC
b. Polyploidy-three or more sets of chromosomes.
c. Triploidy-3 sets of chromosomes (3n) : ABC, ABC, ABC.
d. Tetraploidy-4 sets of chromosomes (4n): ABC, ABC, ABC, ABC.
e. Pentaploidy-5 sets of chromosomes (5n) : ABC, ABC, ABC, ABC, ABC.
f. Hexaploidy (6n), Septaploidy (7n), Octoploidy (8n), etc
2. Aneuploidy - variation in the number of chromosomes of a set. (Reduction in
the normal number of chromosomes. )
a. Monosomics - Loss of one chromosome (2n-1) : ABC, AB.
b. Double monosomics - Loss of 2 different chromosomes (2n-1-1): ABC, A.
b - loss of a pair of homologous chromosomes (2n-2) : AB, AB:
b. Increase in the number of chromosomes (polysomies).
Trisomies - presence of 1 extra chromosome (2n+ 1) : ABC, ABC, A.
Double trisomics - 2 different extra chromosomes (2n + 1 + 1) : ABC, ABC, AB.
Tetrasomics - an extra pair of homologous chromosomes (2n+2): ABC, ABC, AA.
pentasomics (2n+3), Hexasomics (2n+4), Sepiasomics (2n+5), etc. Euploidy
The term euploidy (Gr., eu-true or even; ploid-unit) designates genomes
containing whole sets of chromosomes. The euploids are those organisms which
contain balanced set or sets of chremosomes or genomes in any number, in their
body cells. The euploidy is of following types: The number of chromosomes in a
basic set is called the monoploid number (x). Organisms with multiples of the
monoploid number of chromosomes are called euploid. Eukaryotes normally carry
either one chromosome set (haploids) or two sets (diploids). Haploids and diploids,
then, are both cases of normal euploidy. Euploid types that have more than two
sets of chromosomes are called polyploid.
Polyploidy
Humans are diploid creatures, meaning for every chromosome in our body, there is
another one to match it.
• Haploid creatures have one of each chromosome
• Diploid creatures have two of each chromosome
• Triploid creatures have three of each chromosome
• Polyploid creatures have three or more of each chromosome
They can be represented by n where n equals haploid, 2n equals diploid and
so on. It is possible for a species, particularly plant species, to produce offspring
that contains more chromosomes than its parent. This can be a result of non-
disjunction, where normally a diploid parent would produce diploid offspring, but in
the case of non-disjunction in one of the parents, produces a polyploid.
In the case of triploids, although the creation of particular triploids in species
is possible, they cannot reproduce themselves because of the inability to pair
homologous chromosomes at meiosis, therefore preventing the formation of
gametes. Polyploidy is responsible for the creation of thousands of species in
today's planet, and will continue to do so. It is also responsible for increasing
genetic diversity and producing species showing an increase in size, vigour and an
increased resistance to disease.
The polyploid types are named triploid (3x), tetraploid (4x), pentaploid (5x),
hexaploid (6x), and so forth. Polyploids arise naturally as spontaneous
chromosomal mutations. However, many species of plants and animals have clearly
arisen through polyploidy, so evidently evolution can take advantage of polyploidy
when it arises. It is worth noting that organisms with one chromosome set
sometimes arise as variants of diploids; such variants are called monoploid (1x). In
some species, monoploid stages are part of the regular life cycle, but other
monoploids are spontaneous aberrations.
The haploid number (n), which we have already used extensively, refers
strictly to the number of chromosomes in gametes. In most animals and many
plants with which we are familiar, the haploid number and monoploid number are
the same. Hence, n or x (or 2n or 2x) can be used interchangeably. However, in

certain plants, such as modern wheat, n and x are different. Wheat has 42
chromosomes, but careful study reveals that it is hexaploid, with six rather similar
but not identical sets of seven chromosomes. Hence, 6x=42 and x=7. However, the
gametes of wheat contain 21 chromosomes, so n=21 and 2n=42.
Monoploids
In monoploidy, the monoploid organisms have one genome (n) in their body
cells. When monoploidy occurs in gametes (sperms and eggs) it is termed as
haploidy. Most micro-organisms (e.g., bacteria. fungi and algae); gametophytic
generation of plants (e.g., bryophytes and other plants); sporophytic generation of
some higher angiospermic plants (e.g., Sorghum, Triticum, Hordeum, Datura, etc.)
and certain hymenopteran male insects (e.g., wasps, bees, etc.) have one genome
in their body cells, hence are monoploids. Monoploids are usually smaller and less
vigorous than their diploid prototypes. Characteristically, monoploid plants are
sterile. The reason of sterility is that the chromosomes have no regular pairing
partners (homologous chromosomes) during meiosis, and meiotic products are
deficient in one or more chromosomes. For instance, a haploid in maize (2n=20)
will have 10 chromosomes and the number of chromosomes in a gamete can range
from 0-10. Consequently, considerable sterility will be found in a monoploid maize.
Male bees, wasps, and ants are monoploid. In the normal life cycles of these
insects, males develop parthenogenetically—that is, they develop from unfertilized
eggs. However, in most species, monoploid individuals are abnormal, arising in
natural populations as rare aberrations. The germ cells of a monoploid cannot
proceed through meiosis normally, because the chromosomes have no pairing
partners. Thus, monoploids are characteristically sterile.
Monoploids play an important role in modern approaches to plant breeding.
Diploidy is an inherent nuisance when breeders want to induce and select new gene
mutations that are favorable and to find new combinations of favorable alleles at
different loci. New recessive mutations must be made homozygous before they can
be expressed, and favorable allelic combinations in heterozygotes are broken up by
meiosis. Monoploids provide a way around some of these problems. In some plant
species, monoploids can be artificially derived from the products of meiosis in a
plant's anthers. A cell destined to become a pollen grain can instead be induced by
cold treatment to grow into an embryoid, a small dividing mass of cells. The
embryoid can be grown on agar to form a monoploid plantlet, which can then be
potted in soil and allowed to mature.
Figure a. Generating monoploid plants by tissue culture. Pollen grains (haploid) are
treated so that they will grow and are placed on agar plates containing certain plant
hormones. Under these conditions, haploid embryoids will grow into monoploid
plantlets. After having been moved to a medium containing different plant
hormones, these plantlets will grow into mature monoploid plants with roots,
stems, leaves, and flowers.
Plant monoploids can be exploited in several ways. In one, they are first
examined for favorable traits or allelic combinations, which may arise from
heterozygosity already present in the parent or induced in the parent by mutagens.
The monoploid can then be subjected to chromosome doubling to achieve a
completely homozygous diploid with a normal meiosis, capable of providing seed. It
is achieved by the application of a compound called colchicine to meristematic
tissue. Colchicine—an alkaloid drug extracted from the autumn crocus - inhibits
the formation of the mitotic spindle, so cells with two chromosome sets are
produced. These cells may proliferate to form a sector of diploid tissue that can be
identified cytologically.
Figure b. The use of colchicine to generate a diploid from a monoploid. Colchicine

added to mitotic cells during metaphase and anaphase disrupts spindle-fiber
formation, preventing the migration of chromatids after the centromere is split. A
single cell is created that contains pairs of identical chromosomes that are
homozygous at all loci.
Another way in which the monoploid may be used is to treat its cells basically
like a population of haploid organisms in a mutagenesis-and-selection procedure. A
population of cells is isolated, their walls are removed by enzymatic treatment, and
they are treated with mutagen. They are then plated on a medium that selects for
some desirable phenotype. This approach has been used to select for resistance to
toxic compounds produced by one of the plant's parasites and to select for
resistance to herbicides being used by farmers to kill weeds. Resistant plantlets
eventually grow into haploid plants, which can then be doubled (with the use of
colchicine) into a pure-breeding, diploid, resistant type.
Figure c. Using microbial techniques in plant engineering. The cell walls of haploid
cells are removed enzymatically. The cells are then exposed to a mutagen and
plated on an agar medium containing a selective agent, such as a toxic compound
produced by a plant parasite. Only those cells containing a resistance mutation that
allows them to live within the presence of this toxin will grow. After treatment with
the appropriate plant hormones, these cells will grow into mature monoploid plants
and, with proper colchicine treatment, can be converted into homozygous diploid
plants.
These powerful techniques can circumvent the normally slow process of

meiosis-based plant breeding. The techniques have been successfully applied to
several important crop plants, such as soybeans and tobacco.
The anther technique for producing monoploids does not work in all
organisms or in all genotypes of an organism. Another useful technique has been
developed in barley, an important crop plant. Diploid barley, Hordeum vulgare, can
be fertilized by pollen from a diploid wild relative called Hordeum bulbosum. This
fertilization results in zygotes with one chromosome set from each parental species.
In the ensuing somatic cell divisions, however, the chromosomes of H. bulbosum
are eliminated from the zygote, whereas all the chromosomes of H. vulgare are
retained, resulting in a haploid embryo. (The haploidization appears to be caused
by a genetic incompatibility between the chromosomes of the different species.)
The chromosomes of the resulting haploids can be doubled with colchicine. This
approach has led to the rapid production and widespread planting of several new
barley varieties, and it is being used successfully in other species too.
Diploidy
The diploidy is characterized by two genomes (2n) in each somatic cell of the
diploid organisms. Most animals and plants are diploids. The diploidy is related with
fertility, balanced growth, great vigorosity, adapatability and survivality of the
diploid organisms
Polyploids
The organisms with more than two genomes are called polyploids. Among
plants and animals, the polyploidy occurs in a multiple series of 3, 4, 5, 6, 7, 8,
etc., of the basic chromosome or genome number and thus is causing triploidy,
tetraploidy, pentaploidy, hexaploidy, heptaploidy, octaploidy, respectively. Ploidy
levels higher than tetraploid are not commonly encountered in natural populations,
but our most important crops and ornamental flowers are polyploid, e.g., wheat
(hexaploid, 6n), strawberries (octaploid, 8n), many commercial fruit and
ornamental plants, liver cells of man, etc. Other examples of polyploidy among
plants and animals are following:
A: Examples of polyploidy in plants
The polyploidy is most common among angiosperms and some of
economically important polyploid angiospermic plants are peanuts (Arachis),oats
(Avena), coffee (Coffea), strawberry (Fragaria), cotton (Gossypium), barely
(Hordeum), sweet potato (Ipomoea), apple (Malus), alfa-alfa (Medicago), banana
(Musa), tobacco (Nicolina), plum (Prunus), sugar cane (Saccharum), potato
(Solanum), sorghum (Sorghum), clover (Trifolium), and wheat (Triticum).
A continuous polyploid series has been reported in rose plant. Aeuploid series
of basic number of 7 (monoploid) including diploids (2n= 14), triploids (21),
tetraploids (28), pentaploids (35), hexapolid (42), and octaploid (56) has been
reported in different species of Rosa. Likewise, the genus Chrysanthemum has basic
chromosome number 9 and has a euploidic series of diploid (2n = 18), tetraploids
(4n=36), hexaploids (6n=54), octaploids (8n=72) and decaploids (10n=90) in its
different species.
The genus Solanum has basic chromosome number 12 and has a euploidic
series of diploids (2n=24), triploids (3n=36), tetraploids (4n=48), pentaploids
(5n=60), hexaploids (6n=72), octaploids (8n=96), and decaploids (10n = 120) in
its different species.
Origin of Polyploidy
Different degrees of ploidy are originated by different means. However, two
basic irregular processes have been discovered by which polyploids may evolve
from diploid plants and become established in nature:
(1) Somatic doubling-cells sometimes undergo irregularities at mitosis and give rise
to meristematic cells that perpetuate these irregularities in new generations of
plants.
(2) Reproductive cells may have an irregular reduction or equation division in which
the sets of chromosomes fail to separate completely to the poles at anaphase. Both
sets thus become incorporated in the same nucleus resulting in the doubling of
chromosome number in the gamete (see Gardner, 1912). Thus, a triploid originates
by the fusion of a haploid gamete (n) with a diploid gamete (2n), the later of which
may be originated by irregularities during meiosis.
Likewise, a tetraploid may be originated by the somatic doubling of the
chromosome number or by union of unreduced diploid gametes.
The somatic doubling of genome is accomplished either spontaneously or it
can be induced in high frequency by exposure to chemicals such as colchicine, etc.,
or heat or cold. Other levels of polyploidy are also originated by same methods.
A. Chromosomes B. Diploid gamete C. Haploid gamete D. Triploid

zygote
Induction of Polyploidy
The polyploidy can be induced in common diploid organisms by following methods:
1. Cell generation - In certain bryophytes such as mosses, the polyploidy has
been induced by cutting their diploid sporophytes and keeping the sporophytes in
moist conditions. The cells of the cut ends regenerated threads which were true
protonema and produced diploid gametophytic generation instead of monoploidic
generation.
2. Physical agents - Following kinds of physical conditions induce polyploidy in
plants:
(i) Temperature shocks - Extreme temperature changes some. times result in a
higher frequency of polyploid cells.
(ii) Centrifugation - The centrifugation of seedlings of plants causes polyploidy in
their cells. In Nicotiana, polyploidy has been induced by this method.
(iii) X-rays - The radioactive substances such as radium and X-rays have been
found to induce polyploidy in normal diploid plant cells.
3. Chemical agents - Some chemicals such as colchicine, chloral hydrate,

acenaphthene, veratrine, sulfanil amide, ethyl, mercury chloride.
hexachlorocyclohexane have been reported to induce polyploidy in plants. These
chemical substances when used to dividing diploid cells, they disturb the mitotic
spindle and cause non-segregation of already duplicated chromosomes and thus,
convert the diploid cells into tetraploid cells. The tetraploid cells, likewise, are
converted into different levels of polyploidy
Kinds of Polyploidy
In the realm of polyploids, we must distinguish between autopolyploids,
which are composed of multiple sets from within one species, and allopolyploids,
which are composed of sets from different species. Allopolyploids form only
between closely related species; however, the different chromosome sets are
homeologous (only partly homologous)—not fully homologous, as they are in
autopolyploids.
Allopolyploidy- The prefix "allo" indicates that nonnhomologous sets of
chromosomes are involved.
P1: Species X X Species Y
(AA) ↓ (BB)
(Diploid) (Diploid)
F1: AB
Diploid sterile hybrid
↓ Colchicine
AABB
Fertile amphidiploid tetraploid
The union of unreduced or diploid or polyploid gametes from different diploid or

polyploid species could produce in one step, an amphipolyploid or allopolyploid;
which appears and behaves like a new species. Let A represent a set of genome in
species X, and let B represent another genome in a species Y. The F 1 hybrids of
these species than would have one A genome and another B genome. The F1
diploid but sterile hybrids can be converted into fertile allotetraploids by treating
them by colchicine.
The Russian cytologist, G. D. Karpechenko (1928) first ore. , synthesized a

allotetraploid genus called Rhaphanobrassica from the artificial crosses between
vegetables belonging to different genera, the radish (Raphanus satirum, 20 = 18)
and the cabbage (Brassica oleracea, 2n= 18). The F1 hybrids were diploid and
having the root of cabbage and leaves of radish. They were highly sterile because
of failure of each set of chromosomes to provide sufficient genetic homology to
affect pairing. Among these sterile F1 hybrids, however, he found certain fertile
allotetraploids which contained 36 chromosomes and were named as
Rhaphanobrassica.
Triticum spelta, is a hexaploid wheat which was artificially synthesized in

1946 by E. S. McFadden and E. R. Sears and also by H. Kihara. They crossed an
emmer wheat, Triticum dicoccoides (tetraploid: 2n= 28) with goat grass, Aegilops
squarrosa (diploid; 2n= 14) and doubled the chromosome number in the F1 hybrids
This artificially synthesized hexaploid wheat was found to be similar to the primitive
wheat T. spelta. When the synthesized hexaploid wheat was crossed with naturally
occurring T. spelta, the F1 hybrid was completely fertile, this suggested that
hexaploid wheat must have originated in the past due to natural hybridization
between tetraploid wheal and goat grass followed by subsequent chromosome
doubling.
Aegilops
P1 Trilicum dicoccoides X
squarrosa
(Tetraploid emmer (Diploid goat

↓
wheat) grass)
AABB DD
(2n=14; 7
(2n=28; 14 bivalents)
bivalents)
F1: ABD
(Triploid hybrid)
(2n=21; 21 univalents)
AA BB DD
Synthesized hexaploid
wheat
(Triticum spelta)
Another interesting case of allotetraploidy has been observed·--tn the

production of a rust resistant allotetraploid wheat plant. Common wheat plant,
Triticum vulgare is susceptible to leaf rust, a seriouse disease caused by the fungus
Puccinia triticina. A wild grass of the Mediterranean region, Aegilops umbellulata is

completely resistant to this disease. Sears (1956) have transferred the genes of
rust resistance of A. umbellulata into T. vulgare genome by following method:
He crossed the plants of A. umbellulata with T. dicoccoides and got sterile
hybrid which by treatment with colchicine was transformed into a rust-resistant,
fertile allotetraploid having 21 pairs of chromosomes. The allotetraploid was
crossed to T. vulgare and a fertile, rust-resistant hybrid was produced.
Gossypium hirsutum, the New world cotton plant, is another interesting
example of allopoly ploidy. Old world cotton, Gossypium herbaceum, has 13 pairs of
chromosomes, while American or "upland cotton" also contains 13 pairs of
chromosomes. J. O. Beasley crossed the old world and American cottons and
doubled the chromosome number in the F) hybrids. The allopolyploids thus
produced resembled the cultivated New world cotton and when crossed with it gave
fertile F1 hybrids These results, thus, suggested that tetraploid Gossypium hirsutum
originated from two diploid species, namely G. herbaceum (20=26) and G.
raimondii (2n=26).
Gossypium
X Gossypium raimondii
herbaceum
(American or upland
(Old world cotton;
↓ cotton, 2n=26; 13
2n=26; 13 bivalents
bivalents
F1 hybrid
(2n=26, 26 univalents)
↓ Colchicine
New wold cotton
(Gossypium hirsurum)
Today, allopolyploids are routinely synthesized in plant breeding. Instead of

waiting for spontaneous doubling to occur in the sterile hybrid, the plant breeder
adds colchicine to induce doubling. The goal of the breeder is to combine some of
the useful features of both parental species into one type. This kind of endeavor is
very unpredictable, as Karpechenko learned. In fact, only one synthetic
amphidiploid has ever been widely used. This amphidiploid is Triticale, an
amphidiploid between wheat (Triticum, 2n=6x=42 and rye (Secale, 2n=2x=14
Triticale combines the high yields of wheat with the ruggedness of rye. The below
figure shows the procedure for synthesizing Triticale.
Figure 18-10. Techniques for the production of the amphidiploid Triticale. If the
hybrid seed does not germinate, then tissue culture (lower path) may be used to
obtain a hybrid plant.
In nature, allopolyploidy seems to have been a major force in speciation of

plants. There are many different examples. One particularly satisfying one is shown
by the genus Brassica, as illustrated in Figure 18-11. Here three different parent
species have hybridized in all possible pair combinations to form new amphidiploid
species.
Figure 18-11. A species triangle, showing how amphidiploidy has been important in
the production of new species of Brassica.
A particularly interesting natural allopolyploid is bread wheat, Triticum

aestivum(2n=6x=42). By studying various wild relatives, geneticists have
reconstructed a probable evolutionary history of bread wheat (Figure 18-12).
Figure 18-12. Diagram of the proposed evolution of modern hexaploid wheat, in

which amphidiploids are produced at two points. A, B, and D are different
chromosome sets
Autopolyploids
The prefix "auto" indicates that the ploidy involves only homologous
chromosome sets. Somatic doubling of a diploid produces four sets of genomes of a
tetraploid and likewise, somatic doubling of a tetraploid produces eight sets of
genomes of a octaploid. Union of unreduced diploid or tetraploid gametes from the
same species would accomplish the same result.For example, if a diploid species
has two similar sets of chromosomes or genomes (AA), an autotriploid will have
three similar genomes (AAA), and an autotetriploid, will have four such genomes
(AAAA). Since an autotriploid remains sterile and cannot produce seeds, therefore,
it has great commercial value in producing seedless varities of economical plants.
For example, in Japan, H. Kihara produced seedless watermelons, which
were autotriploids. Common "doob' grass of U. P. and Bihar is an autotriploid. Other
common seedless autopolyploids are grapes, sugarbeet, Banana, etc. In O.
lamarckiana, the giant mutant described by de Vries was later on discovered to be
an autotetraploid. Further, whenever autopolyploids, originate in nature, these
would be eliminated due to natural selection.
The chromosome sets or genomes are identical. The genome formula
(capital letters represent a group of chromosomes that is generally referred to as
the basic genome or chromosome set) is AAA (autotriploidy), or BBBB
(autotetraploidy),etc. Autopolyploids are also called polysomicpolyploids.
Origin of Autopolyploids
Autopolyploids spontaneously occur in the nature in a low frequency and can
be induced artificially using various ways, such as heat and chemical treatments,
decapitation, and selection from twin seedlings. The effective method to obtain
autopolyploids is using colchicine.
Colchicine is a spindle fiber poison or suppressant. It inhibits the spindle
mechanism at mitosis, resulting in multiples of normal chromosome number.
Triploids
Triploids are usually autopolyploids. They arise spontaneously in nature or
are constructed by geneticists from the cross of a 4x (tetraploid) and a 2x (diploid).
The 2x and the x gametes unite to form a 3x triploid. Autotriploids are genetically
equal to trisomics for each chromosome. The three chromosomes will pair as a
trivalent or a bivalent plus a univalent. Chromosome separation from such pairing is
irregular. Daughter nuclei will receive either one or two copies from each
chromosome. Consequently, most of the gametes resulting from autotriploid
individuals do not have balanced chromosome complements and are not viable. If
progeny survives from autotriploids it is mostly an aneuploid. Autotriploids can be
produced by crossing diploids with their corresponding autotetraploids. The high
sterility of autotriploids has been explored in plant breeding. Triploid bananas (2n
= 33) are vigorous but seedless and therefore preferred for food consumption.
Triploid watermelons have only undeveloped seeds. Triploid is also applied in

seedless Citrus cultivars.
Triploids are characteristically sterile. The problem, like that of monoploids,
lies in pairing at meiosis. Synapsis, or true pairing, can take place only between two
chromosomes, but one chromosome can pair with one partner along part of its
length and with another along the remainder, which gives rise to an association of
three chromosomes. Paired chromosomes of the type found in diploids are called
bivalents. Associations of three chromosomes are called trivalents, and unpaired
chromosomes are called univalents. Hence in triploids there are two pairing
possibilities, resulting either in a trivalent or in a bivalent plus a univalent. Paired
centromeres segregate to opposite poles, but unpaired centromeres pass to either
pole randomly as below:
Figure d. Two possibilities for the pairing of three homologous chromosomes before
the first meiotic division in a triploid. Notice that the outcome will be the same in
both cases: one resulting cell will receive two chromosomes and the other will
receive just one. The probability that the latter cell can become a functional haploid
gamete is very small, however, because, to do so, it would also have to receive
only one of the three homologous chromosomes of every other set in the organism.
Note that each chromosome is really a pair of chromatids.
Autotetraploids
Autotetraploids arise naturally by the spontaneous accidental doubling of a
2x genome to a 4x genome, and autotetraploidy can be induced artificially through
the use of colchicine. Autotetraploid plants are advantageous as commercial crops

because, in plants, the larger number of chromosome sets often leads to increased
size. Cell size, fruit size, flower size, stomata size, and so forth, can be larger in the
polyploidy.
Figure e. Epidermal leaf cells of tobacco plants, showing an increase in cell size,
particularly evident in stoma size, with an increase in autopolyploidy. (a) Diploid,
(b) tetraploid, (c) octoploid. (From W. Williams, Genetic Principles and Plant
Breeding. Blackwell Scientific Publications, Ltd.)
Here we see another effect that must be explained by gene numbers. Presumably
the amount of gene product (protein or RNA) is proportional to the number of
genes in the cell, and this number is higher in the cells of polyploids compared with
diploids.
Polyploid plants are often larger and have larger organs than their diploid
relatives. Because 4 is an even number, autotetraploids can have a regular meiosis,
although this is by no means always the case. The crucial factor is how the four
homologous chromosomes, one from each of the four sets, pair and segregate.
There are several possibilities, as shown in figure below.
Figure f. Meiotic pairing possibilities in tetraploids. (Each chromosome is really two

chromatids.) The four homologous chromosomes may pair as two bivalents or as a
quadrivalent. Both possibilities can yield functional gametes. However, the four
chromosomes may also pair in a univalent-trivalent combination, yielding
nonfunctional gametes. A specific tetraploid can show one or more of these
pairings.
Aneuploidy
Aneuploidy is the second major category of chromosome mutations in which
chromosome number is abnormal. An aneuploid is an individual organism whose
chromosome number differs from the wild type by part of a chromosome set.
Generally, the aneuploid chromosome set differs from wild type by only one or a
small number of chromosomes. Aneuploids can have a chromosome number either
greater or smaller than that of the wild type. Aneuploid nomenclature is based on
the number of copies of the specific chromosome in the aneuploid state. For
example, the aneuploid condition 2n−1 is called monosomic (meaning “one
chromosome”) because only one copy of some specific chromosome is present

instead of the usual two found in its diploid progenitor. The aneuploid 2n+1 is
called trisomic, 2n−2 is nullisomic, and n+1 is disomic.
Nullisomics (2n−2)
Although nullisomy is a lethal condition in diploids, an organism such as
bread wheat, which behaves meiotically like a diploid although it is a hexaploid, can
tolerate nullisomy. The four homoeologous chromosomes apparently compensate
for a missing pair of homologs. In fact, all the possible 21 bread wheat nullisomics
have been produced. Their appearances differ from the normal hexaploids;
furthermore, most of the nullisomics grow less vigorously.
The diploid organisms which have lost a pair of homologous chromosomes are
called nullosomics with the genomic formula, 2n-2. The nullosomics exhibit reduced
vigor, fertility and survivality, but, polyploidic nullosomics such as nullosomic
hexaploid what (6n-2) survive to maturity because of the genetic redundancy in
polyploids.
Monosomics (2n−1)
Monosomic chromosome complements are generally deleterious for two main
reasons. First, the missing chromosome perturbs the overall gene balance in the
chromosome set. Second, having a chromosome missing allows any deleterious
recessive allele on the single chromosome to be hemizygous and thus to be directly
expressed phenotypically. Notice that these are the same effects produced by
deletions.
Nondisjunction in mitosis or meiosis is the cause of most aneuploids.
Disjunction is the normal separation of homologous chromosomes or chromatids to
opposite poles at nuclear division. Nondisjunction is a failure of this disjoining
process, and two chromosomes (or chromatids) go to one pole and none to the
other. Nondisjunction occurs spontaneously; it is another example of a chance
failure of a basic cellular process.
Monosomics show the deleterious effects of genome inbalance, as well as

unexpected expression of recessive alleles carried on the monosomic chromosome.
Figure AN 1. The origin of aneuploid gametes by nondisjunction at the first or

second meiotic division.
Trisomics
A diploid cell with an extra chromosome. Basically a diploid with an extra
chromosome of one type, producing a chromosome number of the form 2n + l. The
diploid organisms which have one extra chromosome are called trisomies. They
have the chromosomal formula 2n+1. In a trisomic, one of the pairs of
chromosomes has an extra member, therefore, forms a trivalent structure during
meiosis.During anaphase of meiosis, two chromosomes go to one pole and one
chromosome to another pole and thus, two types of gametes n + 1 and n are
resulted. The trisomy has variable effects on the phenotype of the organism and in
man trisomy of autosome 21 cause mongolism.
In plants, first case of trisomy was investigated in Datura stromonium

(Jimson weed) by
Blakeslee and Belting in 1924. D. stramonium normally has 12 pairs of
chromosomes in the somatic cells, but in a individual they discovered 25
chromosomes (2n + 1). The size, shape and spine characteristic of seed capsule of
this trisomic plant had difference with seed capsule of the wild type. species.
Theoretically, because the complement was composed of 12 chromosome pairs
differing in the genes they carried, 12 distinguishable trisomies were possible in
Jimson weed. Through experimental breeding, Blakeslee and his associates
succecceded in producing all 12 possible trisomies. These were grown in Blakeslee's
garden and each was found to have a distinguishable phenotype that was attributed
to an extra set of the genes contained in one of the 12 chromosomes.
Tetrasomy
When one chromosome of an otherwise diploid organism is present in
quadruplicate, the tetrasomy is resulted. The tetrasomics have the .chromosomal
formula 20+2. During meiosis a quadrivalent is formed by extra chromosomes and
segregation of chromosomes occurs like autotetraploids.
Double Trisomy
In a diploid organism when two different chromosomes are represented in
triplicate, the, double, trisomy is resulted. A double trisomic has the chromosomal
formula 2n+1+1.
The following indicates in details the types of chromosome mutation where whole
genes are moved:
Lecture 21
STRUCTURAL CHROMOSOMAL ABERRATIONS
Structural chromosomal aberrations cause structural abnormalities in

chromosome structure. They alter the sequence or the kind of genes present in
chromosome. These are further classified into four groups based upon whether they
alter the gene sequences, number or location.
Changes in the structure of chomosomes.
a. Loss or addition of segments of chromosomes.
Deletion (deficiency) - Loss of a segment of a chromosome
Duplication - repetition of a segment of a chromosome.
b. Changes in the normal arrangement of genes in the chromosome.
Translocation – Exchange of segments between two non - homologous
chromosomes, resulting in new chromosomes.
Inversion – Change in the linear order of genes by rotation of a section of a
chromosome through 180 degrees.
Gene mutations or point mutations – changes in the nucleotide sequence of a
gene.
a. Deletion
b. Insertion
c. Substitution
d. Inversion
a. Loss or addition of segments of chromosomes
Deletion of a Gene
As the name implies, genes of a chromosome are permanently lost as they become
unattached to the centromere and are lost forever
Normal chromosome before mutation

• Genes not attached to centromere become loose and lost forever
• New chromosome lacks certain genes which may prove fatal depending on
how important these genes are
Duplication of Genes
In this mutation, the mutants genes are displayed twice on the same chromosome
due to duplication of these genes. This can prove to be an advantageous mutation
as no genetic information is lost or altered and new genes are gained
Normal chromosome before mutation

• Genes from the homologous chromosome are copied and inserted into the
genetic sequence
• New chromosome possesses all its initial genes plus a duplicated one, which
is usually harmless
The next page continues looking at these chromosome mutations and mutations
that happen within genes that can prove to be more harmful to the organism at
hand. The following pages also investigates polyploidy in species.
b. Changes in the normal arrangement of genes in the chromosome

Inversion of Genes
This is where the order of a particular order of genes are reversed as seen below
• Normal chromosome un-altered
• The connection between genes break and the sequence of these genes are
reversed
• The new sequence may not be viable to produce an organism, depending on
which genes are reversed. Advantageous characteristics from this mutation
are also possible
Translocation of Genes
This is where information from one of two homologous chromosomes breaks and
binds to the other. Usually this sort of mutation is lethal
• An un-altered pair of homologous chromosomes
• Translocation of genes has resulted in some genes from one of the
chromosomes attaching to the opposing chromosome
II. Gene mutations or point mutations

Alteration of a DNA Sequence
The previous examples of mutation have investigated changes at the chromosome
level. The sequence of nucleotides on a DNA sequence are also susceptible to
mutation.
Substitution
A substitution is a mutation that exchanges one base for another (i.e., a change in
a single "chemical letter" such as switching an A to a G). Such a substitution could:
1. change a codon to one that encodes a different amino acid and cause a small
change in the protein produced. For example, sickle cell anemia is caused by
a substitution in the beta-hemoglobin gene, which alters a single amino acid

in the protein produced.
2. change a codon to one that encodes the same amino acid and causes no
change in the protein produced. These are called silent mutations.
change an amino-acid-coding codon to a single "stop" codon and cause an
incomplete protein. This can have serious effects since the incomplete protein
probably won't function.
Insertion
Insertions are mutations in which extra base pairs are inserted into a new place in
the DNA.
Deletion
Deletions are mutations in which a section of DNA is lost, or deleted.
Inversion
Frameshift
Since protein-coding DNA is divided into codons three bases long, insertions
and deletions can alter a gene so that its message is no longer correctly parsed.
These changes are called frameshifts.
For example, consider the sentence, "The fat cat sat." Each word represents
a codon. If we delete the first letter and parse the sentence in the same way, it
doesn't make sense.
In frameshifts, a similar error occurs at the DNA level, causing the codons to be
parsed incorrectly. This usually generates truncated proteins that are as useless as
"hef atc ats at" is uninformative.
Genetic mutations increase genetic diversity and therefore have an important part
to play. They are also the reason many people inherit diseases.
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What are some types of gene mutations discussed?

What are some types of gene mutations discussed?

Principle of Genetics

This eCourse Developed By

1. History of Genetic 1-9

2. Mendel’s laws of inheritance and exceptions to the laws. 10-28

3. Types of gene action 29-37

4. Multiple alleles 38-43

5. Multiple factor hypothesis. 44-47

6. Quantitative traits - Qualitative traits and differences between them 48-54

7. Cytoplasmic inheritance 55-60

8. Methods of inducing mutations and C l B technique. 61-66

9. Mutation - it’s characteristic features – 67-75

10. Regulation Gene expression 76-82

11. Lac operon and Fine structure of Gene. 83-98

13. Study of chromosome structure, morphology, number and types - 119-135

17. Transcription – Translation - Genetic code and outline of protein 179-202

19. Linkage 216-224

20. Numerical chromosomal aberrations (Polyploidy) 225-246

21. Structural chromosomal aberrations. 247-251

01. HISTORY OF GENETICS

HISTORICAL DEVELOPMENTS IN GENETICS

Year Scientist Contribution

1485 L. da Vinci Recommended the use of lenses for viewing small

1590 Z. Janssen and Produced the first operational microscope.

1668 F. Redi Disproved the theory of spontaneous generation of

1672 Malphigi Classified the tissues.

1674 A.van Leeuwenhoek Improved lens system of microscope by grinding.

1682 N. Crew Described bladders and pores in wood and pith.

1694 J.R. Camerarius Conducted early experiments on pollination and

1700 R. Linnaeus Classified the biological organisms.

1761 J.C. Kolreuter Hybridized various species of tobacco and

1779 C.F Wolff Founder of embryology.

1825 F.V. Raspail Developed the frozen-section technique and used

1835 H. von Mohli Emphasized the importance of protoplasm and

1837 R. Brown Discovered the nucleus in cells of flowering plants.

Year Scientist Contribution

1840 J.E. Purkinj Gave the term “ protoplasm ”.

1845 A. Donne Used photomicroscopy for the first time.

1859 C. Darwin Put forth the theory of natural selection.

1862 Kolliker Used the term “ cytoplasm ”for the living

1865 G. Mendel Developed the fundamental principles of heredity.

1870 W. His Invented the microtome.

1871 F. Meischer Isolated nucleic acids from pus cells.

1873 H. Fol Described spindle and astral rays.

1875 O. Hertwig Studied reproduction in sea urchins and concluded that

1879 W. Flemming Introduced the term “ chromatin ”.

1881 E.G. Balbiani Discovered giant chromosomes in salivary glands of

1882 W. Flemming Coined the term “ mitosis ” .

1883 W. Rouse Proposed that chromosomes contain genes which are

Year Scientist Contribution

1885 A.F.W. Schimper Introduced the term “ plastids ”.

1888 Th. Boveri C oined the term “ centrosomes ”.

1888 W. Waldeyer Coined the term “ chromosomes ”.

1892 O. Hertwig Proposed the protoplasm theory of inheritance.

1892 J. Ruckert Described lamp brush chromosomes in oocytes of shark.

1892 Th. Boveri Described meiosis in Ascaris.