DEFENSIVE MECHANISMS OF PLANTS BASED ON SECONDARY METABOLITES

Diana Ioana Alexan, anul II, Universitatea de Vest Timişoara, Facultatea de Chimie,
Biologie, Geografie, specializarea Biologie

Abstract

The plants are surrounded, in any environment, by biotic and abiotic stressors. In order to
protect themselves from the attack of herbivorous, fungal or bacterial infections, they
synthesize secondary metabolites. The secondary metabolites may be terpenes, phenolic
compounds or compounds containing N and S derived directly from amino acids. The
content of this review about defensive byproducts aims to classify and explain the role and
mechanism of their operation.

Key words: secondary metabolites, terpenes, phenolics, defense mechanism, phytoalexins,

environmental stress.

Introducere

In any ecosystem, plants are exposed to stress and damage caused by biotic and abiotic
environmental factors. Pests such as viruses, bacteria, fungi or consumers, grazing insects and
mammals are the main cause of the decline in the success rate of plant cultures. To protect these
factors, plants synthesize secondary metabolites. In superior plants, secondary metabolites are the
synthesized substances from the primary metabolites (carbohydrates, lipids and amino acids)
(Ramakrishna & Gokare, 2011). These protective substances are: terpenes, phenolic compounds,
alkaloids, sterols, nitrogen and sulfur compounds.

Most commonly, a variety of stress factors attack the plant simultaneously, so secondary
metabolites are not specifically produced against a particular pathogen, but depending on the need
to resist more hazards. Thus, in addition to the defense component with these metabolites, plants
use the formation of barriers to polymers against pathogens or the synthesis of enzymes capable of
degrading the cell wall of pathogens (Hammond et al., 1996). After eliminating a first infection,
plants often acquire immunity to support subsequent microbial attacks (Putnam & Heisey, 1983,
Putnam & Tang, 1986).

Natural plant products protect them against predators and pathogens, against abiotic stress
(UV rays, temperature variations, alkalinity or acidity of the soil, salinity), but also facilitate
communication processes with other plants. Secondary metabolites also contribute to the formation
of plant characteristics such as taste, odor, or specific color (Ravishankar & Venkataram, 1990). On
the other hand, they are unusable for growth and development. Moreover, it is known that the
synthesis of secondary metabolites is very expensive, reducing the growth and propagation
capacity, or that their toxins could affect the plant that produces them and the environment
(Gershenzon, 1994). One way to lower energy costs would be for biosynthesis of defensive
substances to be initiated after pathogens or consumers attack. The risk of this strategy is that the
attack may be too rapid or its effects too devastating in order for the plant to be able to defend itself
in an induced manner.
 Compounds that have a negative effect on the development, multiplication or survival of
other foreign organisms have all the status of toxins, playing a role in defense mechanisms
(Gershenzon & Wittstock, 2002). These may be secondary metabolites that are constitutive
substances, called phytoanticipins, or induced, newly formed as an immune response, called
phytoalexins. Phytoanticipins are considered the first barrier to pathogens, phytoalexins being
subsequently synthesized, a process that may take several days after initiation of the attack
(Grayer & Harborne, 1994).

This article describes the types of secondary metabolites and their role in defending
mechanisms against biotic and abiotic environmental factors. Metabolites can be divided into
various groups, depending on their chemical composition, defensive action, or storage site at the
level of the plant organisms. The chemical composition classifies them in 3 major classes: products
containing terpenes, containing phenols and those containing nitrogen and sulfur.

Terpenes

It is the largest class of secondary defense compounds and acts against the attack of insect
and mammalian herbivores, representing toxins for them. These compounds are composed of 5-C
isopentanoic units, their biosynthesis being made from acetyl-coA and glycolic intermediates
(Gerhenzon et al., 1991, Grayson, 1998, Fraga 1988, Croteas 1988, Loomis and Croteas 1980,
Robinson 1980). Terpenes are divided into 5 subclasses:

1.1. Monoterpenes (C10) are toxic agents for insects. Monoterpene esters, called
pyrethroids, are located in the leaves and flowers of some species of the genus
Chrysanthemum. They have neurotoxin action on wasps, moths or beetles and are
used in commercialised insecticides because of their minimal negative effect on the
environment and mammals (Turlings et al., 1995). Monoterpenes are also present in
coniferous species (fir, pine) accumulated in needles and trunks resin channels. In
these gymnosperms, the terpenes are in the form of: limonene, myrecen, α and β-
pinene (Turlings et al., 1995).

1.2. Sesquiterpenes (C15) are cyclic esters that protect plants of various
pathogens, insects and herbivores (Picman, 1986). A complex of 4 sesquiterpene
phytoalexins was identified in a cotton plant resistant to a Verticillium dahliae
infection (Garas & Waiss, 1986).

1.3. Diterpenes (C20). An example of a substance is the abietic acid, present in

the resinous channels of pines and fruit trees. It chemically discourages insects that
feed themselves by perforating the channels (Bardley et al., 1992). Forbol, the
diterpenic ester found in Euphorbiaceae, produces skin irritation and internal
intoxication in consuming mammals. Diterpenes can also have direct positive effects
on plant functions that contain them. Thus, the chlorophyll phytate increases the
efficiency of the photosynthesis process by optimally fixing CO2 and maximizing
biomass production (Jagendorf, 1967).
 1.4. Triterpenes (C30), represented by steroidal alcohols (sterols), play an
important role in maintaining the permeability of small molecules at the plasma level
as they decrease the movement of fatty acid chains (Mazid et al., 2011). Triterpena
called limnoid is a complex of bitter substances present in citrus. It poisoned
herbivores which attack plants from the Rutaceae family (Mordue & Blackwell,
1993).
1.5. Politerpenes ((C5) n) have high molecular weight. This subclass includes
tetraterpenes. Carotenoid pigments and rubber (polymer up to 15,000 units of
isopentenil) are such tetraterpene, their action providing plant protection and good
metabolic processes. The rubber located in the lattice channels heals wounds
produced at plant organs and protects them from herbivores (Eisner et al., 1995,
Klein, 1987).

2. Phenolic compounds

Phenols are derived from the synthesis of shikimic acid and aromatic amino acids. It is
known that the production of phenolic compounds with a role in plant defense is closely correlated
and directly proportional to the amount of light to which plant organisms are exposed (Chalker &
Fnchgami, 1989). Also, the amount of ozone in the atmosphere supports their biosynthesis
(Saviranta et al., 2010).

2.1. Coumarins are simple phenolic compounds, acting against herbivores and
fungi. Derived from shikimic acid (Murray et al., 1982), they are highly active and have high
antimicrobial potency, protecting plants from bacteria and fungi (Brooker et al., 2008). Their
halogenated derivatives are used in the exploration of new compounds with antifungal
activity due to pesticidal behavior. Thus, simple 7-hydroxy coumarins have defensive action
against the Orobanche cernua parasite, inhibiting its germination and stopping its vascular
communication at the root of the host (Serghini et al., 2001).

2.2. Furano-coumarins are spread within the Umbelliferae family (carve, celery,
etc.) and are not toxins. These substances are activated by the ultraviolet rays of Type A,
penetrate inside the DNA molecule of the pathogens, blocking the transcription process and
ultimately killing the cells (Rice, 1987).

2.3 Lignin: is a phenolic polymer having three monomers: coniferyl, coumaryl

and synapil, the distribution of their individual quantity being different from one plant to
another and even in the same cell wall composition (Lewis & Yamamoto, 1990). The
protection afforded by this substance consists in the hardness it gives to the cell walls of the
plant cell, making it harder for herbivores to feed. Lignin is also indigestible for certain
insects and animals (Mader & Fisher, 1982). Plant lignification is also a response to
infection and injury.

2.4. Flavonoids are phenolic derivatives that do not play the role of toxins, but
support the plant by performing pigmentation and defenses against harmful B-rays.
Blavones and flavonols accumulate in the epidermis, thus absorbing the visible and
uninterrupted wavelengths of UV-B rays (Lake et al., 1983).
 2.5. Isoflavonoids are derived from naringerin, an intermediate of flavonol.
Biosynthesis and their presence in leguminous plants has the role of helping to form the
nitrogen fixation nodules at the root level, made by symbiosis with Rhizobium bacterial
genus (Sreevidya et al., 2006).

2.6. Tannins are phenolic, toxic polymers that have rejecting effects for many
herbivores. Because of the ability of these metabolites to bind proteins, they come in
contact with those in saliva for animal users, and provide an unpleasant, pungent and
astringent sensation at the level of their mouth. Thus grazing livestock such as cattle, sheep
or goats avoids plants with high content of tannins (Oates et al., 1980).

3. Metabolites containing sulfur

Amino acid derivatives are known for their antimicrobial action. This class includes:

3.1. Glutathione (GSH) is a tripeptide and brings a considerable amount of

organic sulfur to the plants. It is considered a mobile sulfur reservoir with a role in plant
growth and development (Kang et al., 2007). Its antioxidant action is present in response to
oxidative stress. Some plants contain specialized cells with enzymatic activity for the
biosynthesis of glutathione and other phytochelatins, all of which are beneficial for the
detoxification of heavy metals (Gutierez-Alcala et al., 2000). The GSH also functions as a
messenger about information about fungal attacks, which is rapidly accumulating. Thus,
information is transmitted to uninfected tissues (Foyer & Rennenberg, 2000).

3.2. Glucosinolates (GSL) are a group of glucosides accumulated in higher plants

of the Brassicaceae family (mustard, cabbage, radishes, etc.), which they protect against
consumers and parasites. These metabolites are released as volatile, toxic and unpleasant
substances for predators (Mitchen, 1992). Volatile odors in GSL are catalysed by
smorinase, resulting from glucose breakdown, pungent products, isothiocyanates and
nitriles that irritate the mucous membranes of the animals.

3.3. Phytoalexins are secondary metabolites synthesized following a fungal or

bacterial attack or pathogen-induced stress (Van Etten et al., 1994). The phytoalexins of the
Brassicaceae family have a special ring structure (indole) and a sulfur atom. They are
synthesized by cruciferous plants (Gross et al., 1993), but are products different from
glucosinolates characteristic of the same families (Harborne, 1999).

3.4. Defensins, thionins and lectins are plant proteins. These do not have the role
of organic sulfur accumulation, just like the above-mentioned metabolites, but act as
inhibitors for a wide range of mushrooms. (Thomma et al., 2002). Defenses have both an
antifungal and antibacterial role. Tionins are responsible for the action against infections, as
observed by Kang and Buchenauer by their accumulation in the cell wall of the wheat
pepper cells infected with Fusarium culmorum. Lectins are proteins that bind to certain
carbohydrates. Their defensive action occurs after ingestion when it blocks the absorption of
nutrients in the digestive system of the consumers (Peumansend Van Damme, 1995).
4. Metabolites containing nitrogen

These metabolites are derived from common amino acids and act as toxins. They
belong to this class: alkaloids, cyanogenic glucosides and non-protein amino acids.

4.1. Alkaloids are spread 20% in vascular plants, especially in herbaceous dicots,
being less common in gynosperm and monocotyledons (Hegnauer et al., 1988). These
secondary metabolites are synthesized from aspartic acid, lysine, tyrosine and tryptophan,
all of which are natural amino acids (Pearce et al., 1991). Alcaloids cause most deaths
among mammals, because the ingestion of plants containing them, such as the Lupinus and
Delphinium genus, has extremely toxic effects (Keeler, 1975). They either interfere with
consumer neurotransmitters or affect vital functions such as protein synthesis or membrane
transport (Creelman & Mullet, 1997).

4.2. Cioanogenic glycosides release hydrogen cyanide (HCN) and H2S, poisonous
volatile substances, when the plants containing them are crushed. They are common
among families Gramineae, Rosaceae and Fabaceae (Seigler, 1991). Examples of such
substances are: amigdaline present in apricot, peach, cherry and durine seeds of Sorghum
bicolor. Black bean (Phaseolus lunatus) is known to be a model plant in the production of
secondary metabolites due to the impressive cianogenicity capacity it has (Ballhom et al.,
2009).

4.3. Non-protein amino acids are either incorporated into proteins, or exist as
defensive products in free form (Johnson et al., 1989). Once ingested, it binds to existing
proteins in the herbivore system, leading to the formation of non-functional proteins (eg.
canavanine binds to arginine).

Conclusion

This paper describes the mechanisms of plant defense against pathogens, consumers and
abiotic factors, based on the biosynthesis of secondary metabolites.

Although this synthesis consumes a lot of the plant's resources, it is extremely useful,
chemically protecting plants from natural hazards, improving some functions, thus contributing to
their longevity.

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