Plant Defences Against Herbivore and Insect Attack

Christopher L Schardl, University of Kentucky, Lexington, Kentucky, USA

Plants deploy a number of defences against herbivore and insect attack. A primary defence is the production of toxins to poison the attackers. Other strategies include the development of thorns and tough unpalatable tissue to deter herbivores, and the recruitment of parasitoids and predators that attack herbivores.

Secondary article
Article Contents
. Introduction . Types of Herbivores and Interactions . Types of Defences . Coevolutionary Dynamics . Plant Secondary Chemistry . Interactions of Chemical Defences . Temporal and Developmental Defences . Biotechnology

Introduction
The story of Adam and Eve links eating a forbidden fruit with death in the cycle of human existence (Genesis 3:1 24). Indeed, to avoid poisoning, herbivores and omnivores such as humans must be cautious in choosing which plants and plant parts to eat. A prevailing hypothesis is that the nearly ubiquitous antiherbivore metabolites in plants were selected by coevolution with herbivores. Likewise, the ability of a herbivore to detoxify or tolerate toxins of food plants is essential for the herbivores survival. Plants may largely escape herbivory if they are imbued with an especially toxic cocktail, although the plants must also have mechanisms for protection from their own toxins. Other defences, such as thorns and tough or unpalatable tissues, and recruitment of parasitoids and predators that attack herbivores, all work in conjunction and synergy with chemical defences. Plants survive only if they are able at least partly to deter or avoid herbivory. Without defences, plants would be driven to extinction, with the consequent extinction of myriad herbivores, predators, parasites and saprotrophs. Thus, the familiar species of our biosphere and the fossil record of longstanding associations of plants with animals indicate a dynamic balance struck between plants and those that eat them. brate toxins that protect their host plants (Bush et al., 1997). A counterexample is bark beetles and their fungal symbionts, which act in concert to feed upon conifers. Three-organism interactions such as highlighted in these examples barely hint at the complexities underlying the food webs for which plants are the base. Such complex webs, which also include predators and parasites of the herbivores, may be highly important for ecosystem stability (Siemann, 1998).

Types of Defences
Chemical defences are ubiquitous and probably essential for plant survival, but other defences are also important, varied, and not always obvious. Plants accumulate tough polymers such as cellulose, lignin, tannins and silicates, which reduce palatability. Also, by minimizing the nutritive value of their tissues plants may force a herbivore (particularly an insect) to consume more. Though this strategy may not seem advantageous, it actually forces the herbivore to ingest larger amounts of plant toxins. Thorns, barbs, stings and sticky resins exuded from resin ducts, lactifers, or trichomes physically interfere with herbivory and trap or kill herbivores. Lectins and proteinase inhibitors produced in response to grazing interfere with digestion. A signicant protective strategy is to recruit as allies parasitoids and predators that attack herbivores. This is especially eective against insects. Plants under attack can release a cocktail of volatile chemicals that act as cues for these allies, which then attack, consume and reduce the populations of herbivores (De Moraes et al., 1998). An advantage to this strategy is that it may require relatively little energy and nutrient resources to produce a highly eective attractant.

Types of Herbivores and Interactions

Any organism that consumes plant tissue is a herbivore. The most intensely studied are vertebrates and insects, but nematodes, molluscs, mites and parasitic microorganisms are also important. These organisms can act together to facilitate the consumption of a plant, or can act against one another in surprising ways that help protect the plant from devastation. Interesting examples of the latter phenomenon include plant mutualisms with ants and with endophytic fungi. Stinging ants directly or indirectly (via aphids) depend on plants for food and are erce protectors of the resource. Symbiotic microorganisms (endophytes) also depend entirely on host plants for nutrition, but often invest considerable resources in anti-insect and antiverte-

Coevolutionary Dynamics
Herbivory often results in signicant reduction in tness of a plant species, so plants come under strong selection to
1

ENCYCLOPEDIA OF LIFE SCIENCES / & 2002 Macmillan Publishers Ltd, Nature Publishing Group / www.els.net

Plant Defences Against Herbivore and Insect Attack

elaborate defences. Then, of course, the herbivores that depend on plants must develop resistances and tolerances to the defences. Thus emerges evolutionary dynamics that take as many twists as there are potential strategies and counterstrategies (Figure 1). As plants utilize combinations of chemical, physical and developmental (phenological) defences, and as they recruit allies, herbivores adapt to greater or lesser degrees to all of these defences. Moreover, adapted herbivores often benet from the original plant defences. Notably a herbivore that has adapted to a class of plant toxins may utilize the same toxins against predators, may appropriate the toxins as olfactory cues for its preferred food plant (kairomones), or may even use them as attractants for mates (pheromones). The coevolutionary dynamics of plant defence and herbivore adaptations necessitate the multifarious nature of plant defences, and probably drive diversication of forms, species and chemistries of plants.

between plant species with similar secondary chemistries (Becerra, 1997). It is dicult to demonstrate that the functions of chemicals produced by plants are to reduce or prevent herbivory. This is because herbivores must have adapted to the defences of at least some plants to at least some degree or they would have become extinct long ago. Nevertheless, the observation that so many plant secondary metabolites are active against so many animals and pathogenic microorganisms strongly suggests a defensive role. Information about some of the more common classes of antiherbivore chemicals in vascular plants is given below.

Alkaloids
Alkaloids include any of a wide variety of chemicals that incorporate amines. Since many neurotransmitters are biogenic amines, many alkaloids have neurotropic eects ranging from extreme bitterness to causing stupor or death. Activities on multiple neuroreceptors are common. An example is the ergot alkaloid class (Figure 2) characteristic of the plant family Convolvulaceae and protective symbionts (endophytes) in grasses. They aect the mammalian central nervous system and interfere with appetite, water homeostasis, temperature regulation, and even fertility. The same alkaloids acting on the peripheral nervous system cause miscarriages and, by reducing blood ow, anoxia and gangrene of the limbs (Bush et al., 1997). Other alkaloids, such as pyrrolizidines and steroidal glycoalkaloids, are directly toxic to cells. These are normally conned to the vacuole of the plant cell to prevent autotoxicity. Herbivores break open the vacuoles during feeding, releasing the toxins that then damage both herbivore and plant tissues. Plants are far more capable than animals of recovering from these insults, so the animals generally lose this chemical battle.

Plant Secondary Chemistry

Plants generally produce chemicals with toxicity to a wide variety of potential herbivores, or which deter herbivores from feeding. These chemicals are almost certainly of major importance for defence against generalist herbivores, and even for restricting the degree of damage caused by specialist herbivores exhibiting some resistance to the chemical defences. Dierent plant groups (e.g. families) tend to have dierent chemical characteristics. Thus, alteration in chemical proles appears to be a slow evolutionary process. Likewise, when herbivores shift diets to new food plants, such shifts are most likely to be

Plants New phenotype

Herbivores Generalist feeding

Terpenoids, steroids and other prenyl derivatives

Metabolites derived from ve-carbon units called isoprenes are extremely varied in their properties and biological activities. Steroid and terpenoid hormones are critical in animal development and signalling, making the receptors for these compounds obvious targets for antiherbivore chemicals. Particularly interesting examples are the phytojuvenoids (Figure 2). These plant compounds mimic juvenile hormones and can thereby stop or cause inappropriate development of herbivorous insects. However, like the alkaloids, some isoprene-derived defences are toxic directly to cells. These include saponins, which are sugar-linked terpenoids, steroids or sterol alkaloids with broad toxicity to animals and microbes. Some plant pathogenic fungi have saponin-detoxifying enzymes that

Protection from generalist feeders (feeding deterrents, attractants for predators, etc.)

New resistance, tolerance or avoidance mechanisms

Adaptive radiation

Adaptive radiation Kairomones Antipredator defences

Elaboration of defences: Additional new defences Adaptive variance of expression and timing of defences

Further adaptations

Figure 1 A simplified scheme for coevolution between plants and their herbivores.

ENCYCLOPEDIA OF LIFE SCIENCES / & 2002 Macmillan Publishers Ltd, Nature Publishing Group / www.els.net

Plant Defences Against Herbivore and Insect Attack

OH O N H
O

N N O

Ergovaline

N H CH 3 O H O
H H O OH

Juvocimene I

O H CH 3 O
HO

O OH O CH 3 O
Juvenile hormone III Myrcene

Labriformidin

O
HO HO

O OH HO

OH OH

Gossypol

Psoralen

Figure 2 Examples of chemical defences of plants against herbivores. Shown are: ergovaline, an alkaloid from the fungal endophyte Epichloe festucae that enhances host resistance to herbivores (Bush et al., 1997); juvenile hormone III from sedge and juvocimene I from sweet basil, phytojuvenoids that interfere with insect development; labriformidin, a toxic cardenolide from milkweed; myrcene, one of the monoterpene components of pine resin; gossypol, a sesquiterpene dimer from cotton; and psoralen, a furanocoumarin from celery that damages DNA when exposed to ultraviolet light. Branches connect carbon atoms except where otherwise indicated, hydrogen atoms are not shown except where needed to indicate stereochemistry, and hexagons with circles indicate aromatic six-carbon rings.

are essential for survival on host plants that produce those saponins (Bowyer et al., 1995). Some of the best known interactions of plants and herbivores involve steroid toxins such as the cardenolides (Figure 2). Because of their pharmacological properties at low doses, sugar-linked cardenolides such as digitalis (actually a cardenolide mixture) are called cardiac glycosides. Cardenolide activity and toxicity is due to their eects on ion transport across cell membranes. In the milkweeds (Calotropis and Asclepias species of the Asclepiadaceae) the cardenolides are dissolved in the sticky terpenoid latex stored in specialized cells called lactifers. Some insects have intricate feeding habits that

minimize exposure to this toxic latex. Furthermore, insects that feed on cardenolide-laden plants often secrete or sequester these toxins as potent deterrents to predators. This appropriation of a plant defence is exemplied by the monarch buttery (Danaus plexippus) whose larvae feed on milkweeds, accumulate cardenolides, and display bright colours that warn away potential predators. Terpenoids and related compounds comprise a huge group of chemicals with numerous biological activities. One of the most apparent classes is the resin-producing terpenoids of pines and other conifers (Figure 2). These resins directly interfere with feeding or even trap herbivores. Also included in this class are important toxins such
3

ENCYCLOPEDIA OF LIFE SCIENCES / & 2002 Macmillan Publishers Ltd, Nature Publishing Group / www.els.net

Plant Defences Against Herbivore and Insect Attack

as gossypol (Figure 2) in cotton and the iridoids common among angiosperms.

Cyanide and glucosinolates

Cyanogens are known from all major plant groups. The cyanogenic glycosides, if unaltered, are relatively nontoxic. However, removal of a sugar moiety by action of a glycosidase enzyme leaves unstable intermediates whose subsequent spontaneous breakdown results in cyanide release. The cyanide poisons respiration and can thereby cause death. Glucosinolates, like cyanogens, are nontoxic forms compartmentalized away from the myrosinase enzymes that cause release of their toxic components. When plant tissue is damaged and vacuoles are ruptured, glucosinolates come in contact with the cytosolic myrosinases that then catalyse their cleavage into a sugar moiety (usually glucose) and an unstable intermediate. The intermediate rapidly breaks down to form sulfate and mustard oils of the form R-SCN (where R represents any of various linked chemical groups), R-CN 1 S, or related structures. Considerable evidence indicates that glucosinolates are toxic to a broad range of mammalian and insect herbivores as well as plant-pathogenic fungi and water moulds (Oomycetes).

Production of more than one toxin, particularly if they are not chemically related, can be an eective counterstrategy to a herbivores resistance against any single toxin. Dierent chemical classes often act in synergy against herbivores. Also, developmental changes in chemical proles can reduce herbivory by restricting the time period that herbivores adapted to the rst defence can continue to feed on the plant. Changes in proles over the age of the plant are frequently observed. Also, herbivory may change the expression of biochemical pathways so that those that are initially prevalent are turned o and those that are quiescent at rst are turned on. Thus, a herbivore may begin to feed on a plant with chemical proles to which it is resistant; the chemical prole may then change to one less favourable for the herbivore.

Temporal and Developmental Defences

Growing plant tissues have high nutritive value and tend to be heavily grazed by herbivores, so they must be well protected. Typically, young tissues have higher concentrations of chemical toxins whereas older tissues are tougher and less palatable. Polymers such as tannins and lignin, terpenoid resins, and silicate deposits contribute to toughness. Also, in some plants the chemical toxins change with development so that herbivores that begin feeding on younger tissues may be unable to cope with the older tissues. Coordinated and rapid ushes of new growth may outpace herbivores. Similarly, mast fruiting can outpace seedeaters. Bamboos are famous for worldwide coordination of their owering and seed production. This is thought to be a trait selected because bamboo seeds are highly nutritious and are not well protected by chemical defences. The sudden abundance of bamboo seeds in mast fruiting would outpace the herbivores and allow a large number to establish a new generation of plants (Janzen, 1976).

Coumarins
Coumarins are derivatives of phenylalanine. As such they are related to the major components of lignin and suberin, and branch early from the pathway to avonoids and tannins. Furanocoumarins such as psoralen (Figure 2) are common in the plant family Apiaceae. Furanocoumarins are phototoxic, forming toxic intermediates in the presence of light. Ultraviolet light in particular causes furanocoumarins to become highly reactive and crosslink DNA. Thus, these compounds tend to have very broad toxicity to animals and microorganisms.

Interactions of Chemical Defences

The advantage of a defence would appear to be obvious: energy and photosynthetic assimilates of the plant remain a resource for survival and reproduction of the plant rather than of the secondary consumers. However, the defence itself requires expenditures of these resources. Thus, if a plant suers relatively little herbivory the cost of the defence may not be recovered in increased tness. Furthermore, if one of several potential herbivores is undeterred by the plants defences, the advantage of the defence might not be realized. Thus, several strategies must be employed that are more sophisticated than simple assault with toxic chemicals.
4

Biotechnology
A major objective of biotechnology is to modify crops and horticultural plants for enhanced resistance to herbivores and pathogens, preferably with the benet to the environment that chemical sprays will be reduced. To introduce new antiherbivore activities it will be necessary to identify all genes required for their production. Therefore, the rst such introductions have been of genes for single antiherbivore proteins. Various subspecies of the bacterium Bacillus thuringiensis produce a variety of d-endotoxins (Bt toxins) with broad or narrow activities against insects (Hofte and Whiteley, 1989). The rst report of Bt toxin in transgenic plants was in 1987 (Vaeck et al., 1987). Now signicant acreage of soya beans, cotton and corn are

ENCYCLOPEDIA OF LIFE SCIENCES / & 2002 Macmillan Publishers Ltd, Nature Publishing Group / www.els.net

Plant Defences Against Herbivore and Insect Attack

projected to be composed of such transgenic plants. An important concern is that overuse of particular Bt toxins in transgenics, or excessive reliance on Bt toxins in general, will select resistant insects (Tabashnik et al., 1998). The concerns point to a well-founded but sometimes ignored principle: reliance on individual defences against potentially devastating herbivores is dangerous, so multiple and varied defences should be sought and employed. Reliance on single proteins like Bt toxins is very limiting. Secondary metabolites will continue to be important where they occur naturally and potentially where they can be introduced by plant transformation. However, it is a daunting prospect to identify and move multiple genes for defensive chemicals from one plant or endophyte into a distantly related plant. Nevertheless, the observation that coevolutionary relationships between plants and herbivores tend to track chemical relationships (Becerra, 1997) suggests that expression of new chemical classes in transgenic crops would be highly eective against their specialized herbivores. Though transformation of plants with new defensive genes is potentially powerful, it is unwise to opt routinely for this approach. Furthermore, an abundance of other possibilities are available or yet to be discovered. Deployment of parasites, pathogens or endophytes against herbivores, and identifying novel pest resistance genes in wild or regional germplasms, are two approaches with considerable promise. Keeping in mind the value of multiple strategies, a thorough understanding of the multifarious defences naturally employed by plants is crucial when devising such defences for agricultural and horticultural plants. After all, in nature many remarkable

and often subtle defence mechanisms underlie the survival of diverse plants and the organisms that depend on them.

References
Becerra JX (1997) Insects on plants: macroevolutionary chemical trends in host use. Science 276: 253256. Bowyer P, Clarke BR, Lunness P, Daniels MJ and Osbourn AE (1995) Host range of a plant pathogenic fungus determined by a saponin detoxifying enzyme. Science 267: 371374. Bush LP, Wilkinson HH and Schardl CL (1997) Bioprotective alkaloids of grassfungal endophyte symbioses. Plant Physiology 114: 17. De Moraes CM, Lewis WJ, Par PW, Alborn HT and Tumlinson JH (1998) Herbivore-infested plants selectively attract parasitoids. Nature 393: 570573. Hofte H and Whiteley HR (1989) Insecticidal crystal proteins of Bacillus thuringiensis. Microbiological Reviews 53: 242255. Janzen DH (1976) Why bamboos wait so long to ower. Annual Review of Ecology and Systematics 7: 347391. Siemann E (1998) Experimental tests of eects of plant productivity and diversity on grassland arthropod diversity. Ecology 79: 20572070. Tabashnik BE, Liu YB, Malvar T et al. (1998) Insect resistance to Bacillus thuringiensis: uniform or diverse? Philosophical Transactions of the Royal Society of London B: Biological Sciences 353: 17511756. Vaeck M, Reynaerts A, Ho fte H et al. (1987) Transgenic plants protected from insect attack. Nature 328: 3337.

Further Reading
Coley PD and Barone JA (1996) Herbivory and plant defenses in tropical forests. Annual Review of Ecology and Systematics 27: 305335. Rosenthal GA and Berenbaum MR (eds) (1992) Herbivores: Their Interactions with Secondary Plant Metabolites, vols III. San Diego, CA and London: Academic Press.

ENCYCLOPEDIA OF LIFE SCIENCES / & 2002 Macmillan Publishers Ltd, Nature Publishing Group / www.els.net