Cyanogenesis in Higher Plants and Insects

Matthias Lechtenberg, University of Muenster, Muenster, Germany

Cyanogenesis describes the ability of plants to store cyanogenic glycosides, which, on tissue damage, undergo hydrolysis with the release of toxic hydrogen cyanide. This defensive mechanism is widely distributed in plants and also occurs in some insects.

Secondary article
Article Contents
. Introduction . Structures . Cyanogenesis: Degradation of Cyanogenic Glycosides . Related Structures . Distribution . Biogenesis in Plants . Toxicology

Introduction
The term cyanogenesis describes the ability of organisms to liberate free prussic acid after the breakdown of hydrogen cyanide (HCN)-storing compounds (so called cyanogens) by catalysis of cleaving enzymes. Often the disruption of tissue or wounding initiates this process by abolition of the compartmentation of substrates (cyanogenic glycosides, cyanogenic lipids) and enzymes (b-glycosidase, hydroxynitrile lyases, esterases). Cyanogenic compounds are mostly regarded as defence compounds. Cyanogenesis has been described for over 2500 species of higher plants and lots of examples are known from insects. Many basic foodstus contain cyanogenic glycosides or breakdown products.

. Plant Insect Interaction

cyclopentenyl glycine group and the nicotinic acid group.

Figure 2 shows one prominent example of each group with

the amino acid precursor.

Cyanogenesis: Degradation of Cyanogenic Glycosides

The action of specic plant or insect b-glucosidases on cyanogenic glycosides leads to fairly unstable cyanohydrins which may decompose enzymatically (catalysed by a hydroxynitrile lyase) or nonenzymatically (depending on the pH) into HCN and a corresponding carbonyl compound. This process is called cyanogenesis. Figure 3 shows the decomposition of linamarin. Cyanogenesis involves large amounts of HCN liberated after the breakdown of cyanogens; the occurrence of low amounts of prussic acid is due to the normal metabolism of plants, for instance during the formation of ethylene or the action of horseradish peroxidase on amino acids.

Structures
Figure 1 shows the general formula for cyanogenic glyco-

sides. All known cyanogenic glycosides are derivatives of a-hydroxynitriles (cyanohydrins). These unstable compounds are stabilized by b-glycosidic bonded sugars or sugar chains. In all documented cases b-d -glucose is the rst sugar attached to the aglycone. As the R1 and R2 residues are often dierent, two epimeric forms are probable. Usually, both forms are known from natural sources but they seldom occur in the same plant (insect) or even in related species (Nahrstedt, 1992). Today, more than 60 cyanogenic glycosides are known from higher plants (Seigler and Brinker, 1993). The structures are classied into biogenetic groups according to their (in most cases putative) precursor amino acids. Six major groups can be derived: the phenylalanine group (including cyanogenic glycosides bearing a meta-hydroxylation at their aromatic ring), the tyrosine group, the valine/isoleucine group, the leucine group, the
CN C Sugar O R1 R2

Related Structures
Closely related to the cyanogenic glycosides of the leucine group is the small group of cyanogenic lipids. Four types are known today. Instead of glycosylation, in cyanolipids the a-hydroxy group of the cyanohydrin is estericated with fatty acids. A further structurally related group consists of the noncyanogenic nitrile glucosides. Often, the misleading term cyanoglucosides is used. These compounds are bglucosides of b- or g-hydroxynitriles. After hydrolysis of the glucosidic linkage, no free cyanohydrins arise. Nevertheless, these nitriles seem to be able to liberate HCN under certain conditions. Thus sarmentosin epoxide (structurally related to cyanogenic glycosides of the valine/isoleucine group) or osmaronin epoxide (related to the leucine group) are responsible for the weak cyanogenesis of Sedum
1

Figure 1 General formula for known cyanogenic glycosides.

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Cyanogenesis in Higher Plants and Insects

1. Phenylalanine group 2. Tyrosine group

3. Valine/isoleucine group

4. Leucine group

5. Cyclopentenyl glycine group

6. Nicotinic acid group

N N C H OGlc

OGlc OCH3 C

Linamarin N C OGlc N H C OGlc N C OGlc

GlcO HO N CH3 O

R Prunasin (R= H) Taxiphyllin (R= OH) Lotaustralin Heterodendin Deidaclin

Acalyphin

OH

OH

H2N Valine

H2N O OH O OH O OH HOOC H2N R Phenylalanine (R= H) Tyrosine (R= OH) Isoleucine Leucine Cyclopentenylglycine H2N H2N N Nicotinic acid

Figure 2 Prominent examples of each biogenetic group of cyanogenic glycosides (blue) with amino acid precursors (red).

OH HO HO O O OH CH3 CN CH3

sarmentosum (Crassulaceae) and Osmaronia cerasiformis (Rosaceae) respectively. The cyclohex(en)ylcyanomethylene glucosides form the largest group of noncyanogenic nitrile glucosides and are putatively derived from tyrosine. A third group consists of two compounds usually discussed together with the cyanogenic glycosides: the free cyanohydrins 4-glucosyloxy-mandelonitrile and its 4-Ocaeic acid ester (trivial name: nandinin), both apparently derived from tyrosine.

(1) H2O OH HO CH3 CN + CH3 HO HO O OH OH

Distribution
Cyanogenesis has been recorded in all major vascular plant taxa in at least 550 genera and 130 families. As most reports are based on simple qualitative tests with HCN test strips (e.g. FeiglAnger or picrate test papers), in most cases the responsible cyanogenic principle remains undetermined. Among the division Pteridophyta, the ferns accumulate cyanogenic glycosides of the phenylalanine group. Within the Spermatophyta, in members of the gymnosperms only taxiphyllin (compare with Figure 2) has been reported to date. Within the Magnoliophytina, cyananogenesis occurs in many families. The dicots are very heterogeneous with

(2)

O CH3 + HCN H3C

Figure 3 Enzymatic hydrolysis of linamarin: (1) b-glucosidase; (2) hydroxynitrile lyase.

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Cyanogenesis in Higher Plants and Insects

regard to the reported structures, whereas cyanogenic members of the monocots are characterized mainly by tyrosine-derived glycosides. Sometimes cyanogenic and acyanogenic phenotypes of the same species occur. This polymorphism has been well studied for white clover (Trifolium repens). The expression of cyanogenesis in white clover is controlled by two independent gene loci. Cyanogenesis has been described in bacteria, cyanobacteria, microalgae and fungi. It is also known in arthropods: the occurrence of cyanogenic glycosides and noncyanogenic nitrile glycosides has been reported in Lepidoptera and Coleoptera.

optimization of its detoxication. Other examples for prominent cyanogenic food plants are lima beans, ax seeds, bamboo shoots, sorghum, bitter almonds, passion fruits and apricot kernels (Nahrstedt, 1993; Jones, 1998).

Plant Insect Interaction

As mentioned above, cyanogenic glycosides are considered to be defence compounds for plants and insects. An interesting example of a herbivore that feeds on a cyanogenic plant and uses cyanogenic glycosides as protecting agents is found in lepidopterous insects (Insecta: Lepidoptera). Some moths of the genus Zygaena are able to accumulate linamarin and lotaustralin. The larvae feed on Lotus corniculatus, which contains the same compounds. Incorporation experiments have shown that the larvae are able to synthesize both linamarin and lotaustralin, or, alternatively, may sequester them from their host plant.

Biogenesis in Plants
Among the biosynthetic studies of cyanogenic glycosides, the biosynthesis of dhurrin (2S-b-d -glucopyranosyloxy-2(4-hydroxy)phenylacetonitrile) in Sorghum bicolor is one of the best investigated examples. l-Tyrosine is the biogenetic precursor of dhurrin. Two steps in the biosynthesis of dhurrin are catalysed by multifunctional membrane-bound cytochrome P450 enzymes. The rst (P450tyr) catalyses the conversion of tyrosine to Z-phydroxyphenylacetaldoxime, the second (P450ox) the conversion of Z-p-hydroxyphenylacetaldoxime to p-hydroxymandelonitrile. In vivo, this cyanohydrin is converted into the cyanogenic glycoside dhurrin by a soluble glucosyltransferase (Mller and Seigler, 1999; Selmar, 1999).

References
Jones DA (1998) Why are so many food plants cyanogenic? Phytochemistry 47(2): 155162. Mller BL and Seigler DS (1999) Biosynthesis of cyanogenic glycosides, cyanolipids, and related compounds. In: Singh B (ed.) Plant Amino Acids: Biochemistry and Biotechnology, pp. 563609. New York: Marcel Dekker. Nahrstedt A (1992) The biology of the cyanogenic glycosides: new developments. In: Mengel K and Pilbeam DJ (eds) Nitrogen Metabolism of Plants, pp. 249269. Oxford: Clarendon Press. Nahrstedt A (1993) Cyanogenesis and foodplants. In: van Beek TA and Breteler H (eds) Phytochemistry and Agriculture Proceedings of the Phytochemical Society of Europe, pp. 107129. Oxford: Clarendon Press. Seigler DS and Brinker AM (1993) Characterisation of cyanogenic glycosides, cyanolipids, nitroglycosides, organic nitro compounds and nitrile glucosides from plants. In: Dey PM and Harborne JB (series eds) Methods in Plant Biochemistry, vol. 8. Waterman PG (ed.) Alkaloids and Sulphur Compounds, pp. 51131. London: Academic Press. Selmar D (1999) Biosynthesis of cyanogenic glycosides, glucosinolates and nonprotein amino acids. In: Wink M (ed.) Biochemistry of Plant Secondary Metabolism, pp. 79150. Sheeld: Academic Press.

Toxicology
The cyanogenic glycosides are potential toxins because of their ability to liberate HCN after hydrolysis. Thus symptoms of acute toxicity of cyanogenic glycosides after ingestion of cyanogenic plant material correlate with those of an acute HCN intoxication. Sometimes intact cyanogenic glycosides show only low acute toxicity because of their slow hydrolysis under the conditions in the gastrointestinal tract. Additionally, detoxication mechanisms of humans and animals are able to detoxify up to 60 mg HCN per h. Nevertheless raised plasma levels of the human detoxication products rhodanide and cyanocobalamin may lead to severe disease. Thus the daily consumption of even subacute amounts of cyanogenic glycosides with cyanogenic food plants leads to chronic cyanide intoxication. The most important cyanogenic food plant is cassava (Manihot esculenta), which provides energy to more than 500 million people, and great eorts are made in the

Further Reading
Ballantyne B and Marrs TC (eds) (1987) Clinical and Experimental Toxicology of Cyanides. Bristol: Wright. Lechtenberg M and Nahrstedt A (1999) Cyanogenic glycosides. In: Ikan R (ed.) Naturally Occurring Glycosides, pp. 147191. Chichester: John Wiley. Vennesland B, Conn EE, Knowles CJ, Westley J and Wissing F (eds) (1981) Cyanide in Biology. London: Academic Press.

ENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Macmillan Publishers Ltd, Nature Publishing Group / www.els.net