Secondary Metabolites

&
Plant Defence

Pankaj Kumar Pant

Lecturer
Dept. Of Biotech. & Allied Sciences
 Secondary Metabolites
• Secondary metabolites are generally those
chemicals that have no significant role in
plant’s growth and development.

• Plant secondary metabolites differs from

primary metabolites in being their specified
occurrence.
• Secondary metabolites are also known as the
Natural products or Secondary Products.
 Role of Secondary metabolites
• Protection against the different organisms.

• Serves as the attractants for the insects

pollinators.

• Functions as the plant-plant competition and

plant -microbe interection.
 Classes of Secondary metabolites
• There are generally three major classes of
secondary metabolites-

1.Terpenes
2.Phenolics
3.Nitrogen containing compounds
 Terpenes or Terpenoids
• Terpenes are derived from the five carbon
compounds known as the Isoprene units.

• The Isoprene is the basic structural unit of the

terpenes.

• Thus, the terpenoids are occasionally known

as the is Isoprenoides.
 Classification of terpenoides
• There are different classes of terpenoides based on
to the number of carbon units, these are –
1. Monoterpenoids( with 10 carbons)
2.Sesqueterpenoides (with 15 carbon units)
3.Diterpenes(with 20 carbon atoms)
4.Triterpenoides(with 30 carbon atoms)
5.Tetraterpenoides(with 40 carbon atoms)
6.Polyterpenoides( {C5}n)
 Terpene Biosyenthesis
• Terpenes are generally synthesized from
primary metabolites by –

- Mevalonic acid pathway

- Methylerythretol phosphate pathway(MEP)

 Mevalonic Acid Pathway
• Acetyl Co-A is the main precursor.

• Three molecules of the Acetyl-CoA are utilized

in stepwise manner to yeild the Mevalonic
acid.
• The mevalonic acid is then phosphorylated,
decarboxylated and dehydrated to form
Isopentenyl diphosphate (IPP).
 Methylerythretol Phosphate pathway

• MEP is yet another pathway leading towards

the synthesis of the Isopentenyl diphosphate
IPP.

-The pathway generally involves the use of

glycolysis intermediates.
- This pathway is operated in the chloroplast
and plastids.
 IPP LEADS TO TERPNOIDS
• IPP generally present along with it’s isomeric
form called Dimethylallyl diphosphate (DPP).

• Both the IPP and DPP are the active

constituents of the terpene biosyenthesis.
- IPP and DPP reacts to form the Geranyl
diphosphate (GPP)
• GPP joins another molecule of the IPP to form
the Farnesyl diphosphate(FPP).
• Farnesyl diphosphate is the main precursor of
the all the sequeterpenes.
• Addition of yet another IPP lead to the
formation Of
Geranylgeranyldiphosphate(GGPP)
 Terpenes And Their Role
• Terpenes are generally toxic in nature and are
generally feeding deterrent to many plant
feeding insects.

• Pyrethroides, Saponins,Phytoecdysomes
essentialoils, are some of the important
terpenoides.
• Pyrethroides- A monoterpenoid ester.
• Found among the members of chrysanthemum
species and are generally good sources of the
insecticides commercially available.
• Essential oils- Essential oils are generally
mixture of volatile monoterpenes and
sesqueterpenes, lend a specific smell to the
foliage.
- Essential oils has the insect repellant acitivities
• Azadirachitin is a non volatile limonoide { a
group of triterpenoide}.

Phytoecdysomes- isolated for the first time from

Podophyllum vulgare.
-It has the resemebelence with the molting
hormone.
• In the insect it thus erupts the molting and
other necessary developmental processes.

Cardiniloides- Generally the group of glycosides

that taste better and are extremely toxic to
the higher animals.
-They generally have the dramatic effect on to
the heart rate by influencing Na/K ion
channel.
- Digitalis have been a good source of the
cardinolides for the treatment of the cardiac
disease.

Saponin
- a steroid and triterpenoide glycoside.
- Posses soap like properties.
- Posses the lipid soluble and water soluble
elements
- Saponin works by disturbing the sterol uptake
from the digestive system.

- Disturbs the membranes after entering the

blood stream.

- Forms the complex with the sterols.

 Phenolic Compounds
• Phenolics are characterised by the presence of
the 6 membered benzene ring along with the
alcohol (–OH).
• Phenolics provides –
-Mechanical support for the pollination.
-Protection against the herbivores and
pathogens.
-Protection against the harmful radiations.
 Biosyenthesis of Phenolics
• There are basically two general pathways are
involved –

1. Shikimik acid pathway.

2. Melonic acid pathway.
ж Melonic acid a pathway is found in fungi and bacteria and is less
common in higher plants.
 Shikimik Acid Pathway
• Shikmik acid pathway generally involves the
conversion of the simple carbohydrate
precursor to “Aromatic amino acid”
Phenylalanine generally.

• Animals generally posses no Shikmik acid

pathway thus, the aromatic amino acid are
essential nutrients in the diet.
 From Phenylalanine to Phenolics
• The most generalised way to form the
phenolics generally involves the removal of
the NH3 to form the Cinnamic acid.
• The above reaction generally involves the use
of the enzyme Phenlyalanine ammonia Lyase
i.e (PAL).
• PAL Is the most studied enzyme in plant
secondary metabolite production.
• PAL generally occupies the position in
between the primary and secondary
metabolism.
• PAL activity generally increases during the
environmentel,nutrient,light and fungal
infections.
• There are cases in he plant where there are
several copies of the PAL are present.
• The reactions further involves the addition of
the hydroxl groups and other substituents.
• The Trans-Cinnamic acid,p-Coumaric acid, and
their derivative so formed are simple phenolic
compounds called phenyl-propanoids.
• These simple phenolics have property to get
activated in presence of light.
• Furanocoumarins-
- Posses the property to resist the phototoxicity.
- Under the influence of sunlight i.e UV-A(320-
400nm) the furanocoumarin gets activated.
- Activated furanocoumarin attach it’self to the
double helix of the DNA and bind to the
cytosine and thymine thus blocks the
transcription and repair pathways.
 Allelopathy

Phenolic compounds like “Caffeic acid” and

“Ferulic acid” generally lead to the inhibition
of seed germination in nearby species a
phenomenon known as the Allelopathy.
• Lignin-
- Highly branched phenyl-propanoid
compound.
- The lignin is generally bound covalently to the
polysaccharide and the cellulose.
- Thus difficult to isolate & so structure not
been correctly elucidated.
- Made up of three phenyl-propanoid phenols
- Coniferyl
- Coumaryl
- Sinapyl
• These components are added enzymatically
that involves the use of the free radicals.
Structural analysis generally showed the
C-O-C, C-C bonding in the three dimensional
orientation.
• Lignin works as a protective agent in plant by-
1. Providing the physical strength that repels the
herb feeder.

2. By binding to the protein and cellulose and so

reducing their digestibility in their feeders.

3. By accumulating at the site of wounding and

infection.
 Flavinoids
• The largest class of the phenolics.

• The structure generallly consists of the 15

carbons.

• Out of which 12 present as the two benzene

ring and the rest three are present as the
bridge carbons.
 FLAVONOIDE GROUPS
• Generally there are four major classes of the
flavnoides-
1.Anthocynin
2.Flavones
3.Flavonols
4.Isoflavones
 Anthocynins
• There are generally two types of the plant
pigments-
1.Carotenoides- Yellow,red,orange colored
2.Flavonoides- With a range of color
-Anthocynins are glycosides with sugar at the
position 3.
-Anthocynins without the sugar are known as
the Proanthocynidines.
Flavones and Flavonols as Phtoprotectants

• Flavones and Flavonols functions as to have

protection against the UV-B(280-320 nm).

• These pigments generally accumulates at the

epidermal cells and so absorb the harmful
radiation and allows only the photosyenthetic
radiations.
 Isoflavones
• Isoflavnoides generally posses the ring B is
shifted.

• Isoflavones are generally the antimicrobial

agents.
• Found mostly in the legumes.
• They generally posses the anti-cancer and
anti-estrogenic activity.
 Tannins
• Tannins is a category of plant phenolic that
generally been utilized for the leather
syenthesis.

• Tannin generally bind to the collagen protein

increasing their resistance to the heat, water
and microbes.
 Classes of Tannins
• Tannins are generally been classified in two
categories-
1. Condensed tannins.
2.Hydrolyzable tannins.
- Condensed tannins- Made from the
polymerization of the flavonoid units.
- Found in woody plants, they could be hydrolyzed
to their anthocynidines, thus are known as pro-
anthocyanidins.
• Hydrolyzable Tannin- They are generally
heterogeneous polymers containing the
Phenolic acids specially the Gallic acid and the
simple sugar.

• They are generally much more easily

hydrolyzed; needs dilute acid.
• The Tannin have the property to bind the
proteins nomspecifically thereby inhabiting their
proper biological action.
• In the gut of many herbivores the tannin is
hydrolyzed to their respective quinoid form.
• Quinoid forms are generally strong electrophiles
that reacts with the nucleophiles –NH2 and –SH
of the protein.
• The Tannin also inactivates the herbivore
digestion by forming the complex aggregates
that are difficult to digest.

• However some of the Rodents and Rabbits are

much resistant to the Tannin toxicity as they
posses the proteins with a high Proline
content.
Alkaloids; N containing compounds
• Alkaloids are group of compounds that are
alkaline at the.
• Contains the N atom that is generally
protonated.
• Protonated N lead to a characterstic positive
charge to the alkaloids.
• Alkaloids are generally derived from some of
amino acids i.e Lysine ,Tyrosine and Tryptophan.
• Stychrine, Conine, Atropine are the classical
examples of the Alkaloids.
• Alkaloids generally effects the nervous system
specially chemical transmitters, ,membrane
transport,protein syenthesis and many other
enzyme activities.
 Cynogenic glycosides
• Cynogenic glycosides are the one of the
compounds of Alkaloid category.
• Found commonly in the Legumes and Grasses.
• Cynogenic glycosides are not posinous in
themselves but they breaks down into the
components that are poisonous.
• Cynogenic glycosides generally releases the
poisonous gas HCN.
 Steps involved in Cynogenic glycoside
Breaking
• The Cynogenic glycosides are generally break
down into toxic components in two steps-
1.The sugar is cleaved by glycosidase produces the
α-Hydroxynitrile or cynohydrin.

2.The cynohydrin is cleaved by Hydroxynitrile lyase

to liberate the HCN.
THE HCN GENERALLY BINDS TO THE METALLOPROTEIN LIKE CYTOCHROME
OXIDASE,A KEY ENZYME IN RESPIRATION.
• Plant are generally resistant to their own
Cynogenic glycosides because the enzyme are
separated in different compartments.

• Cynogenic glycosides are generally feeding

detterents.
 GLUCOSINOLATES
• Glucosinolates is another category of the
Alkaloid Glycosides.
• Found commonly in the Brassicaceae family.
• The sugar moiety is generally attached to the
Sulphur group
• Generally the compounds are responsible for
the typical smell of some vegetables like
Cabbage, Broccoli and Redishes.
 Glucosinolate breakdown
• Glucosinolate breakdown is a two step process-
1. The first step involves the breaking of Sugar-
Sulphur bond to form a Aglycone,the step is
catalyzed by enzyme Thioglucosidase or
Myrosinase.
2. Depending on to the condition the resulting
aglycone is rearranged and loses the sulphate
and gives the Isothiocynates and nitriles.
• Isothiocynates and nitriles are generally the
defensive compounds due to their toxicity and
feeding repellent property.

• Like cynogenic glycosides the enzymes are

seprated in different compartments.
 Non protein amino acids as defence
components
• There are many amino acid in addition to the
standard 20 amino acids these amino acids are
called Unusual amino acids.
• These amino acids are generally not incorporated
into the protein and present in free form.
• Canavanine-
- A unusual amino acid and a close analogue of the
Arginine.
- Canavanine sometime mistakenly
incorporated into the protein.
- This canavanine is identified by the herbivore
enzyme that usually bind to the Arginine
t-RNA.
- The unusual appearance of the Canavanine in
place of Arginine lead to inactivation of
protein.
 Defence responses in plants
• There are generally two types of plant responses to
the pathogenic attacks-
1. Constitutive Defence response .
2. Induced defence response.

Constitutive plant response-

-Are the mechanism that are always present.
-Responsible elements are generally stored
compounds or their precursors.
• Induced plant defence response-
-Are initiated only after the actual damage
occurs.
-Thus, is a kind of rehabilitation action.
 Elicitors;Induction of series of action

• The insect derived compounds that are

responsible for the wound response by plant.
• A generalized group have been identified acts
as the elicitors, these compounds known
commonly as Fatty acid-Amino acid.
• The biosynthesis of these compounds
generally depends upon the plant as source of
fatty acid and insect as amino acid source.
• The fatty acid component generally includes
the Linoleic acid (18:2) and Linolenic acid (18:3).
• Plant part when ingested by insect then,
enzyme in the gut generally conjugated the
insect derived amino acid commonly Glutamine
with plant derived fatty acid.
• Further processing of this conjugate is found in
some insect species.
 Elicitors Induces; Jasmonic acid
• Octadecanoid pathway- when a elicitor is
introduced then it results in a signal transduction
network
• The major signal transduction network is
generally Ocatdecanoid pathway (Chloroplast and
peroxisome).
• In the plants a major response to the insect attack
is formation of jasmonic acid formation through
the well known Octadecanoid signaling pathway.
 Role of Jasmonic acid
• Jasmonic acid generally plays it’s part by
Increasing the transcription of the genes
involved in the defence.
A CLOSE OBSERVATION OF ARABIDOPSIS MUTANT LACKING THE ACTIVE
GENE SHOWED A PRONOUNCED ATTACK BY INSECT THAT DON’T ATTACK
ARABIDOPSIS GENERALLY.
Plant proteins and their role In defence
• There are some plant proteins that have some
role in defence-
α Amylase inhabitors- Generally blocks the action
of the starch digesting enzyme α-amylase.
Lectins- Lectins are generally carbohydrate
binders, they binds to the carbohydrate
associated with the membrane and inhabit the
nutrient uptake.
• Proteinase inhabitors-
- Proteinsase inhabitors are generally best known
anti-digestive proteins.
- Found in case of the legumes tomatoes and
other plants.
- After entering the herbivore digestive system
they binds to the active site of the protein
hydrolyzing enzymes, thereby inhabiting them.
 Systemic Defence
• In case of tomatoes the rapid accumulation of
the