INTRODUCTION

The Chemotaxonomy is concerned with the application of chemical characters in

relation to taxonomy. This rapidly expanding discipline of plant taxonomy also

called as chemosystematics or comparative phytochemistry.

TAXONOMY:

It is a branch of science, which deals with the laws governing the classification of

plants.

The classification of plants as

Division : (Thallophyta, Bryophyta, Pteridophyta)

Subdivision : Gymnospermae and angiospermae

Class : Monocotyledonae and Dicotyledonae

Sub class : Archichlamydeae

Order : Rosales, generally ending with ales

Sub order : Rosineae generally ending with ineae

Family : Rosaceae, generally ending with aceae

(compositae, graminae and labiatae exception)

Subfamily : Rosoideae, generally ending with – oideae

Tribe : Roseae, generally ending with eae

Sub-tride : Rosinae generally ending with inae

Genus : Rosa

Species : Rose indica

The major taxonomic categories used in taxonomy, in the ascending order are

species, genus, family, order, class and division.

Chemotaxonomy: The application of chemistry to systematics is

chemotoxonomy or chemical taxonomy. The concept that plant can be classified

on the basis of their chemical characters is not new and it has a long been of

practical value.

Eg: Aroma from the crushed leaves and fruits. (which is due to presence of

characteristic essential oils) of the Apiaceae (Umbelliferae) and Lamiaceae are the

characteristic points for the identification of the members of these 2 families and 2

sub-familes of the Asteraceae. The Tubiflorae (latex vessels are absent) and

Liguliflorae (latex vessels are present) are distinguished on the basis of the

presence and absence of latex respectively (the both subfamily belonging to

companulales order).

As compared to morphological characters, chemical constituents are more

definable and of more fundamental significant for the plant taxonomy.

THE ORIGINS OF CHEMOTOXONOMY:

The earliest attempts to correlate chemistry with the phylogenetic level of

development was that of Helen Abbott (1886). Abott made some interesting

comments: “the theory of evolution in plant life is best illustrated by chemical

constituents of plant form”. The first successful attempt to combine chemical and
morphological evidence in the study of a single genus was the work of Baker and

Smith (1920) on the essential oils of Eucalyptus. According to the morphological

and chemical data collected on 176 Eucalyptus species , they divided the genus

into 3 groups differing in both morphological structure and chemical constituents.

The concluded that primitive species are those which have feather-veined leaves

and high Pinene content in their essential oils, while more advanced type have

intermediate venation and contain pinene and Cineole.

According to their system, the most advanced taxa show butterfly wing venation

and contain oils with Phellandrene, Piperitone, and geranyl acetate

The work of Baker and Smith was the first comprehensive chemotaxonomic

morphological study of a complex genus and in general their conclusions have

been a significant contribution to the taxonomy of eucalyptus.

Types of chemotaxonomy:

Chemotaxonomy is further classified as

1) Descriptive taxonomy

2) Dynamic taxonomy (based on biosynthetic pathway)

3) Serotaxonomy (specific proteins)

4) DNA characters (DNA hybridization)

The recent growth of chemotaxonomy as a popular field of research is largely due

to the development of rapid analytical techniques, such as chromatography and

electrophoresis. These methods permit the screening of many species or samples

in a reasonably short time. Taxonomists typically have many species to study and

little time for laborious chemical analysis. If such fast methods have been

available in the early years of the century chemotaxonomy might perhaps have

been a favourite field of research much earlier.

The most recent developments in the technology of automatic protein

‘sequenators’ have exciting possibilities for chemotaxonomy. By the use of the

new equipment, the sequences of amino acids in proteins may be compared

relatively quickly.

Principles of chemotaxonomy:

The principle of “chemical taxonomy” is the investigation of the distribution of

chemical compounds, or groups of biosynthetically related compounds in series of

related or supposedly related plants. Chemical contributions to the classification

of plants are based on their chemical constituents; i.e. on their molecular

characteristics.

These characteristics are genetically controlled and have the advantage over

morphological ones, that they can be very exactly described in terms of definite

structural and configurational chemical formula. In this way it helps in

understanding of their biosynthesis, which is a matter of fundamentals systemic

importance.

The plants, which are similar morphologically and chemically, should have the

same ancestor. This is not always true. For instance both crotalaria

(Leguminosae) and Senecio (Compositae) produce pyrrolizidine alkaloids by

similar pathways, but these genera are not related morphologically.

Moreover, “chemical divergence” caused by mutations can also give rise to large

differences in the production of secondary chemical constituents, such chemically

abnormal plants creates great difficulties to chemotaxonomist.

CHEMOTAXONOMIC INVESTIGATIONS:

Chemotaxonomic studies should include the investigation of the pattern of

compounds occurring in plants and preferentially in all the various individual parts

of plant such as the bark, wood leaves, roots, cuticles and seeds. The chemical

constituents generally vary considerably from one organ to another. It is always

dangerous to draw taxonomic conclusion from the occurrence or non-occurrence

of a single compound in a single part of plant. Although compounds of

considerable taxonomic value may be found in any part of a plant, it is reasonable

to assume that the most important ones occur in phylogenetically old,

conservative, little specialized organs. Thus before any important conclusions are

drawn about the presence or absence of a particular compound adequate sampling

of a large number of members of the species at different stages of development and

growing in different environment should be made.

Six main stages can be recognized in a chemotaxonomic investigation:

a) Choice of group, taxonomic survey and sound sampling.

b) Choice, mastery and modification of suitable chemical techniques in a pilot

survey.

c) Full analysis of all material

d) Interpretation and comparison with data from all other sources.

e) Adjustment of classification as necessary

f) Treatment of any evolutionary relationship indicated by the new data.

INFORMATION FROM MOLECULES

The taxonomists and evolutionists are primarily concerned with the molecule as a

carrier of information. Zuckerkandl and Pauling have defined information –

carrying molecules as semantides. Deoxyribonucleic acid (DNA) is now widely

recognized as the physical basis of the genetic code – the information necessary to

construct a new individual. This information is similar in similar organisms and

hence also in plants in similar species and similar genera. DNA is a primary

source of taxonomic information, as well as being the blue print for development

and differentiation. Evolutionists believe that similarity between organisms is

normally proportional to the number of ancestors which they share, and to the age

of their most recent common ancestor. Therefore DNA is also a primary source of

information relating to phylogeny. Zuckerkandl and Pauling characterized it as a

primary semantide. Ribonucleic acid (RNA) is a secondary source of this

information (a secondary semantide) while the translation of the code into

sequences of amino acids provides the taxonomist with tertiary semantides in the

form of proteins.

Applications of chemotaxonomy:

Plant Division: Chemical characters always advantageous over morphological

characters being more precise and easily detectable. Application of chemical

methods in higher plant taxonomy is produced by work on tea.

1) China tea (Camellia sinensis subspecies, sinensis)

2) Assam tea (Camellia sinensis subspecies, Assamica)

3) Ortificial Assam x China F, (China Hybrid tea).

4) Southern form

Tea is generally classified as camellia sinensis. Using chromatography Robert

studies the phenolic content of the young leafy shoots of the tea plants. It was seen

that, China tea and Assam tea were from the same species but the Southern form of

tea from a different species namely Camellia taliensis.

Related Genera having similar enzyme system producing Analgous

metabolites:

The pyrrolizidine alkaloids (senecio) are analogous with quinolizidine (Lupin),

although their precursors are Ornithine and Lysine respectively but they have

similar enzyme system. However it has also observed in that both type of alkaloid

have been found to occur in botanically related genera eg. “Senecio alkaloids”

reported to occur in Crotalaria (Leguminosae, Papiolionatae) and “Lupin

alkaloids” have been isolated from Lupinus and Cytisus (Pepilionatae) both

belonging to Genistae genera, which also cover crotalaria.

Complex metabolic products are good chemotaxonomic marker:

The occurrence of complex secondary metabolites like bioflavonoids are helpful in

the classification of medicinal plants.

The Biflavonoids are plant polyphenols widely distributed in the plant kingdom

with over 4000 different structures. Structurally they are divided into several

subclasses; Flavanol, Anthocyanin, and Chalcones.

The biflavonyls have been reported in leaves of gymnosperms. It has been

reported that the biflavonyls are absent in the genera of pinaceae, hence the

inability to synthesise biflavonyls can be used as a characteristic of the order

pinales hence the complex structure compounds like biflavonyls are very important

for the chemical classification of gymnosperms.

Chemotaxonomy also gives the pylogenetic relationships

Investigation of new drugs.

Helps in the controversy of drugs or where there is a problem of interpretation of

drug classification.

CHEMOTAXONOMY AND ECONOMIC AND MEDICAL BOTANY:

Phytochemists and economic botanists are often looking for new sources of

important chemical compounds (e.g. Colchicines, quinine, reserpine, digitoxin).

Generally, they will be most successful when they follow the lead of natural

classification supplemented by phytochemical knowledge. i.e. if they use the

chemotaxonomic approach.

Chemotaxonomy is also useful for prevention and treatment of plant poisoning of

animal and man. If poisoning is caused by a plant species related to Colchicum

autumnale, for instant by Gloriosa superba from a bunch of flowers or by

bulbocodium vernum cultivated in a garden, it will be wiser to suspect Colchicine

poisoning and to act accordingly.

The same holds for many types of plant contact dermatitis.

Tuliposide-A and its allergenic product of hydrolysis, tulipalin-A, not only occur

in all species of the large genus Tulipa, but also in related liliaceous genera such as

Erythronium, Gagea.

Very recently, Hausen et al reported that species of Lilium and Allium triquetrum

but not Asparagus officinalis, contain moderate amounts of 1-tuliposide A. These

authors suggested that breeding of the above mentioned ornamental liliaceous taxa

for high tuliposide-B and low tuliposide-A content could result in mildly allergenic

cultivators retaining a reasonable disease resistance.

INTRODUCTION TO THE FLAVONOIDS:

It is estimated about 2% of all carbon photosynthesized by plants (about 1 x 10 9

tonns / annum) is converted to flavonoids.

Flavonoids thus constitute one of the largest groups of naturally occurring phenols.

They are virtually ubiquitous in green plants and as such are likely to be

encountered in any work involving plant extracts. For this reason it is important

that chemists biochemists, plant physiologists and biologists generally know how

to recognize, isolate and identify these natural products in all their many forms.

Flavonoids are the largest group of phenolic compounds. Flavonoids occur, either

as free molecules or as glycosides. They occur ubiquitously in ferns and fern allies

(pteridophyta) Conifers (Gymnosperms), dicots and monocots. The dicot families

like compositae, leguminosae, polygonaceae, Rutaceae, Umbelliferae etc. contain

a large number of flavonoids. Flavonoids have been reported from some green

algae also.

Chemically, flavonoid show a 15 carbon skeleton (C 6 – C3 – C6) which consists of

2 phenyl rings connected by 3-carbon bridge.

Most of the flavonoids have a carbonyl function situated at one end of the bridge.

Chalcones and Dihydrochalcones represent the two classes of flavonoids in which

the 3-C bridge in “open”. But in remaining classes the three carbon bridge is a

part of heterocyclic ring, involving a phenolic group on the adjacent ring.

Flavones represent the common flavonoid structure having Carbon ring (carbonyl

group present on heterocyclic ring). Flavones show 2-phenyl -chromone

structure.

Although pure flavones are colourless, the yellow colour of their derivatives

increases with rise in pH and number of OH groups. Isoflavones are isomers of

flavones where C ring and B ring are joined through C 3 rather than C2 (as in

flavones).

The ‘A’ benzene ring arises by a condensation of acetate units, while the ‘B’

benzene ring is a product of a normal pathway of phenolic biosynthesis (the

Shikimic acid pathway). The flavonoid nucleus is normally linked to sugar, so

forming a water soluble glycoside. Most flavonoids are found in the vacuole of

the plant cell.

Guide to the distribution of flavonoid types in nature

Flavonoids are characteristic constituents of green plants with the exception of

algae and hornworts. They occur in virtually all plant parts including leaves, roots,
wood, bark, pollen, nectar, flowers, berries and seeds. In the few recorded cases of

flavonoids being found in animals, for example in the beaver scent gland, propolis

(bee secretion) and in butterfly wings, it is considered that the flavonoids originate

from the plants upon which the animals feed rather than being biosynthesized in

situ (Harborne, 1967). The distribution of flavonoid types within the largest

group, the angiosperms, is further elaborated. Another important feature of the

distribution of flavonoids in plants is the strong tendency for taxonomically related

plants to produce similar types of flavonoids. Thus, useful information about the

flavonoid type likely to be encountered in a plant under investigation may often be

gained by reference to the literature pertaining to previous flavonoid studies of

related plants, e.g. from the same genus or family.

Chemotaxonomic evidence from plant phenolics:

No category of small molecule which has been examined from a taxonomic

viewpoint has proved so popular as that of the phenolic compounds. The main

reason for this popularity is that they are quickly and simply extracted from the

plant material, easily separated by paper chromatography, and fairly readily

identified by location reagents.

Rather little was known about phenolics until paper chromatography was applied

to them. It was then soon clear that in plants they are present in considerable

numbers, diversity, and quantities, although no single compound is found

everywhere, as are the protein amino acids for example.

CHEMICAL STRUCTURE OF PHENOLICS

1) Flavones: Flavones in the root from which the word flavonoid is derived. The

commonest flavones are apigenin and luteolin.

These are widespread in the leaves and flowers of angiosperms.

Flavone itself is rarer, but occurs as a white mealy farina on the leaves and stem of

certain species of primula.

2) Flavanones: Flavanones differ from flavones in lacking the double bond in the

2,3 position. Though quite widespread they are especially common in particular

families, such as Rosaceae, Rutaceae, Leguminosae and compositae. They are

also found in the fern family polypodiaceae and in the pinaceae.

Naringenin is quite common. Pinocembrin is found in Pinus.

3) Isoflavones and Isoflavonoids: Isoflavones are isomeric with flavones, having

the B ring attached at the 3 position instead of the 2 position. Isoflavonoids have

the 2,3-double bond reduced, so that they are related to isoflavones in the same

way that flavanones are related to flavones.

Isoflavones are most characteristic of subfamily Lotoideae of Leguminosae,

though a few have been found elsewhere.

Orobol is a typical isoflavone. Ferreirin is a simple isoflavonoid from Ferreira

(Leguminosae). Isoflavonoids are also best known from Leguminosae.

4) Flavonols: These very common compounds differ from flavones in having a 3

hydroxyl substituent. Numerous flavonols have been identified from plant tissue.
Kaempferol, Quercetin and Myricetin are by far the commonest.

5) Anthocyanidins: These materials are normally found as the aglycone (non –

sugar moiety) of naturally occurring glycosides with glucose, galactose, arabinose

and other mono and di-saccharide sugars.

In other cases the anthocyanidin may be attached to an organic acid (acyl group0

such as parahydroxy benzoic acid.

Acid hydrolysis of the colourless glycosides often called leuco – anthocyanins,

releases the coloured anthocyanidin aglycone. Unlike all the flavonoids so far

considered the anthocyanidins lack the carbonyl group at position 4.

Anthocyanidins include most of the natural red and blue pigments.

The other flavonoids are mostly pale yellow, cream on colourless (anthoxanthins).

Cyanidin, delphinidin and pelargonidin are the 3 commonest anthocyanidin.

They occur in leaves and flowers of most plants. Though only a few

anthocyanidins are known there is much diversity owing to variation in the

glycosides which they can form.

6) Chalcones and Aurones: There are the anthochlor pigments – yellow flower

pigments which turn orange – red in the presence of ammonia. There is little

structural variation in this group, which can be exemplified by the chalcone –

butein and the aurone – sulphuretin.

Reduction of the double bond in a chalcone gives rise to a dihydrochalcone, a few

of which have been found in nature.

Phloretin occurs as the glycosides phloridzin in several species of Malus

(Rosaceae).

In chalcones and dihydrochalcones the heterocyclic C3 ring is open.

7) Biflavonyls: Biflavonyls are dimers of the flavone apigenin.

Amentoflavone is the simplest member.

It is interesting that the biflavonyls seem to be especially common in the

gymnosperms.

FUNCTIONS OF PHENOLIC COMPOUNDS

Plant phenolics used to be regarded as inert end products of metabolism, but

radioactive tracer techniques show that they are capable of considerable

interconversion. They are common in all higher plants and many of the lower

groups. It seems unlikely that they have no function, eventhough they are not

involved in the fundamental metabolic processes. They are found, in varying

quantities, in most tissues of the plant.

The functions of the flower pigments in pollination are fairly clear: insect and bird

pollinators are attracted by coloured petals. There has been a tendency, in the

evolution of flower pigments of insect – pollinated plants, towards bluer and more

stable colours.
Some phenolics may have no other function than to be intermediates in the

biosynthesis of more complex plant products. Coniferyl alcohol is probably

involved in the production of lignins. Gallic acid is a precursor for hydrolysable

tannins.

Phenolic glycosides, which are more soluble than the parent phenolic aglycones,

may serve as a means of transport of the elaborated phenolic residues about the

plant. Possibly the glycosides are a more stable form in which phenolics may be

stored.

Some phenolics may be implicated in growth and development. Work on pea

flavonoids showed that changes in content were rapid enough to be causally

related to growth changes. Decreased growth of internodes was related to

increased synthesis of Kaempferol conjugates. These are known to be co-factors

for the oxidative destruction of indole acetic acid (IAA). In peach buds the

flavanone naringenin preserves dormancy by activation IAA oxidase.

Moewus showed that the sex of algal gametes was determined by certain

flavonoids.

The notion that plant phenolics often have the function of deterring would be

predators. Bate – Smith comments on the unpleasant astringency of developing

pears. The taste is due to the presence of tannins in the unripe fruits. When the

sees are fully developed, it is selectively advantageous that the fruit be edible and

attractive. Presumably the high tannin concentrations in young fruits are a

‘device’ which discourages premature seed dispersal. Tannins combine with skin

proteins and render them resistant to putrefaction; hence their value in the leather

industry. Allard showed inhibition of tobacco mosaic virus by tannic acid.

Inhibition of mechanical transmission of virus has been demonstrated, apparently

caused by a reaction between the virus and a polyphenol. It seems probable that

many phenolics inhibit the germination of fungal spores, and may also be

implicated in the host reactions which contain infections which have already

begun. Substances which inhibit further development of fungus in hypersensitive

host tissue have been defined as phytoalexins. Possibly some of the normal

phenolics of plant tissue may act as precursors for phytoalexins, such as pisatin in

peas. Pisatin is accumulated by pea tissue in the presence of a wide range of

fungal species.

Functions of phenols in wood seems to be connected with decay resistance.

Phenolic compounds are particularly concentrated in the heartwood. Most of then

have been shown to have antifungal properties.

Muller and Chou discuss a role of secreted phenols as water soluble phytotoxins,

preventing establishment of competing vegetation.

Phenolics thus have a wide range of possible functions in plants. Those which are

concerned with more basic processes of growth and development may be found to
occur widely. Those involved with protection may be as varied as the

environmental and ecological pressures which they may help to minimize.

TAXONOMIC VALUE OF PHENOLIC STUDIES

Early applications of evidence from studies on phenolics to taxonomic problems

are found in the work of ZBate – Smith. He established background knowledge of

the distribution of the commoner plant phenolics. Abandoning the flower

pigments as too variable, he concentrated on the phenolics of seeds and vegetative

tissues. One of his early findings was that presence of leuco-anthocyaninswas

correlated with the woodiness of the plants.

Over 60% of ‘woody families’ examined contained leuco-anthocyanins, whereas

they were detected in only 15% of ‘herbaceous families’. Exceptions to the

general rule included the herbaceous family Primulaceae, which was rich in leuco-

anthocyanins. A similar but less striking correlation was shown between woody
habit and the presence of flavonols. Methoxycinnamic acids were shown to be

correlated with the herbaceous habit. These correlations seem merely to reflect the

difference in physiology between woody and herbaceous plants.

Bate-Smith also demonstrated the amazing richness of phenolic variation in plants.

It is the discontinuous distribution of rarer phenolics and the correlated occurrence

or absences of commoner ones which offer potentially valuable evidence to a

taxonomist.

Even widespread phenolic types such as leuco-anthocyanins, phenolic acids and

flavonols may therefore have taxonomic value in specific cases. It seems probable

that greater taxonomic interest will attach to be occurrences and distribution of rare

phenolic aglycones and glycosides. Considering the flavonoid glycosides in leaves

of all five known members of Eucryphiaceae.

Distribution in Eucryphia
South America Australasia
Cordifolia Glutinosa Moorei Milliganii Lucida
Caryatin + + - - -
Az 3-galactoside - + - - -
Az 3-diglucoside + + - - -
Az 3-arabinosylgalactoside + - - - -
Azakeatin (Az) + + - - -
Unidentified flavonone (?) + + - - -
Quercetin 3-galactoside + + - + -
Quercetin 3-rhamnoside + + - - -
Quercetin 3-diglycoside + + - - -
Quercetin 3-triglycoside - - + - -
Dihydroquercetin3-glycoside(s) - - - + -
It was found easy to distinguish the South American from the Australasian species,

though all five species are morphologically very similar.

The rare glycoside luteolin – 5- glucoside has been found in the umbelliferae

genera Chaetosciadium and Torilis.

These genera appeared superficially different in the key characters of fruit

morphology. The new biochemical evidence of similarity between them correlates

with indications from fruit anatomy, and draws attention to the fact that they are

the only genera in the tribe Caucalineae with 12 chromosomes.

In the Coniferae, Erdtman has produced biochemical comparisons of orders,

families and genera. He has been able to distinguish very easily between the

subgenera Diploxylon and Haploxylon of Pinus. The former contained the

flavanones pinobanksin and pinocembrin, with stilbene derivatives called

pinosylvins. The Haploxylon pines had in addition flavones and

dihydropinosylvins.