Terpenoids: Lower

Jeffrey B Harborne, University of Reading, UK

Lower terpenoids are formed in plants either from acetyl-coenzyme A via mevalonate or more directly from glucose via 1-deoxy- D-xylulose. Monoterpenoids are produced from the condensation of two isoprene units, and sesquiterpenoids from three. Lower terpenoids are secreted in glandular hairs on leaves or in the scent glands of flowers. They provide many of the fragrant odours of plants. The best-known sesquiterpenoid is abscisic acid, an important plant hormone involved in the opening and closure of leaf stomata.
. Iridoids

Secondary article
Article Contents
. Biosynthesis . Monoterpenes

. Sesquiterpenoids . Abscisic Acid as a Plant Growth Substance . Sesquiterpene Lactones: Occurrence and Biological Properties

Biosynthesis
All terpenoids are formed by head-to-tail condensation of 5-carbon isoprene precursors, dimethylallyl diphosphate (DMAP) and isopentenyl diphosphate (IPP). Monoterpenoids are formed from two such units (Figure 1) and sesquiterpenoids from three such units. These lower terpenoids are relatively volatile compared to higher terpenoids, which require four or more isoprene units for their formation. At one time, it appeared as though the two key C5 intermediates, DMAP and IPP, were always formed via mevalonate from the condensation of three acetyl-coenzyme A units, with subsequent loss of one carbon atom by decarboxylation. More recent experiments have indicated the existence of an alternative route to DMAP and IPP

from glucose via the key intermediate 1-deoxy-d -xylulose and its 5-phosphate. This nonmevalonate pathway is particularly associated with the leaf plastids but can occur elsewhere. Tracer feeding experiments have conrmed that both pathways to C5 intermediates operate in the biosynthesis of the lower terpenoids (Lichtenthaler, 1999). In monoterpenoid biosynthesis, the rst 10-carbon intermediate, formed from the union of DMAP and IPP, is geranyl diphosphate (Figure 1). This may undergo further enzymatic modication to yield acyclic monoterpenes such as geraniol itself, a principle in the oil of geranium, lemon grass and rose, and linalool, from the oil of coriander. However, many monoterpenoids are monocyclic (e.g. limonene) or bicyclic (e.g. a-pinene) and therefore require a cyclizing enzyme to complete their biosynthesis.

O HO CH3 OPP Dimethylallyl diphosphate HO HO + HO HO CH3 OH

Mevalonate OPP Isopentenyl diphosphate 3 acetyl-coenzyme A

1-Deoxy-D-xylulose

Glucose

OPP cyclase

HO

Geranyl diphosphate

Limonene

trans-Carveol

Carvone

Figure 1 Biosynthesis of monoterpenoids formation of carvone in Mentha spicata.

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Terpenoids: Lower

Currently more than 20 monoterpenoid cyclases have been identied and are characterized in terms of the cyclic product formed. A series of enzymatic oxidations and reductions then come into operation to complete the synthesis of the many dierent known monoterpenes. In a typical case, carvone, a major component of the oil of the mint Mentha spicata, is formed from geranyl diphosphate by a cyclase that converts it to limonene, and this is oxidized in a two-stage process via trans-carveol to carvone (Figure 1). A nal step in the biosynthesis of monoterpenoids in fruits may be the conjugation through a free hydroxyl group (as in geraniol) with glucose to give a glucoside. Such a glucoside may undergo catabolism during the ripening process, with the release of the free monoterpene, as part of the attractive odour of that fruit. The biosynthesis of iridoids or monoterpene lactones follows the same general pathway as for monoterpenes, but several further enzymatic steps may be necessary. The biosynthesis of the iridoid aucubin in Plantago major is illustrated in Figure 2. A key step is the oxidation of geranyl diphosphate to 10-hydroxygeraniol. This is subsequently further oxidized to the corresponding diketone, 10oxogeranial, which rst cyclizes to 8-epiiridodial and then lactonizes to 8-epideoxyloganic acid. Several further modications, together with the linking of a glucose residue, produce the nal product, aucubin. The biosynthesis of sesquiterpenoids begins with the formation of farnesyl diphosphate from the condensation of geranyl pyrophosphate and IPP. This then undergoes a variety of enzymatic modications to produce the many sesquiterpenoids known in nature (Charlwood and

Banthorpe, 1991). Here, attention will be concentrated on the biosynthesis of abscisic acid, a sesquiterpene acid with growth-regulating properties. Ever since its discovery in 1964, it has been assumed to be formed from farnesyl diphosphate, according to the scheme shown in Figure 3. However, an alternative pathway involving the degradation of a carotenoid, violaxanthin, and the intermediacy of 2-cis-xanthoxin has been proposed more recently. While it is possible that both of these routes to abscisic acid operate in plants, the latest evidence from feeding and other experiments indicates that the degradative route via 2-cisxanthoxin is the favoured one (Parry, 1993).

Violaxanthin (C40 carotenoid)

O HO 2-cis-Xanthoxin

CHO

OPP

OH O Abscisic acid CO2 H

Farnesyl diphosphate

Figure 3 Alternative pathways of abscisic acid biosynthesis.

Geranyl diphosphate

OH OH O

10-Hydroxygeraniol CO2 H

10-Oxogeranial

8-Epiiridodial

O OGlc 8-Epideoxyloganic acid

HO

O OGlc Aucubin

HO

Figure 2 Biosynthesis of iridoids formation of aucubin in Plantago major.

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Terpenoids: Lower

Monoterpenes
Monoterpenes, together with sesquiterpenes and aromatics, are components of plant essential oils. They tend to accumulate in members of certain families, such as the Labiatae, Pinaceae, Rutaceae and Umbelliferae, from which they are commercially produced. They are employed in avouring food and in perfumery. Some monoterpenes are ubiquitous in their natural occurrence and can be found in small amounts in the volatile secretions of most plants. Monoterpenes are particularly associated in the plant with specialized secretory structures, such as oil cells, glandular hairs and resin ducts. Their main functions in plants are for attracting pollinators to owers, and most sweet-smelling oral scents are likely to contain a variety of monoterpene constituents. For example, limonene is a dominant odour constituent of Citrus ower, whereas geraniol is dominant in some rose petals. The role of monoterpenes as leaf constituents is less clear-cut, but there is increasing evidence that leaves rich in monoterpenoid constituents are protected from herbivores. For example, camphor occurs in the leaves of white spruce, Picea glauca, and feeding experiments show that this compound repels snowshoe hares from browsing. Other mammals such as red deer avoid feeding on terpene-rich leaves. Some Australian fauna, e.g. possums and koala bears, are able to eat eucalypt leaves containing limonene, cineole, piperitone and terpineol because they have become specially adapted. They avoid any deleterious nutritional eects by absorbing the terpenes from the stomach and small intestine and then detoxifying them via the liver (Harborne, 1993). With over 700 known structures, monoterpenoids can be classied into four arbitrary categories: acyclic, monocyclic, bicyclic and irregular. Typical acyclic monoterpenes are geraniol itself, biosynthetically the simplest (see Figure 1), and then linalool, nerol and citronellol. The last occurs both free and in ester form in many plant oils, e.g. in Boronia citriodora (Rutaceae), and is especially prized by the perfumery industry. Monocyclic terpenes include limonene, a-terpineol and terpinolene, together with the two typical mint leaf oils, menthol and menthone. Bicyclic monoterpenes, which are regularly present in plant essential oils, include a-pinene, b-pinene, borneol and thujone. a- and b-Pinene occur richly in the oleoresin of Pinus palustris and other Pinus spp., and are obtained commercially from these bark oleoresins. Among irregular monoterpenes are the tropolones of gymnosperm heartwoods, e.g. g-thujaplicin, while the pyrethrins of Tanacetum cinerariifolium have extended use in agriculture for their insecticidal properties. Much eort has been expended on the synthesis of monoterpenoids in plant cell culture and, after many frustrating failures, some successes have been achieved. For example, hairy root cultures of ginger, Zingiber ocinale, will produce geraniol and neral in reasonable

yield. Likewise, shoot organ cultures of Pelargonium fragrans will form a-pinene, b-pinene and sabinene (Charlwood et al., 1990). Also, the gymnosperm Pinus radiata in callus culture synthesizes a- and b-pinene at similar levels to those in the intact needles (Banthorpe et al., 1986). Besides their widespread occurrence in plants, monoterpenoids are occasionally found in insects as pheromones and as defence agents. Pine bark beetles use myrcene, ipsdienol and verbenone as aggregation pheromones. The beetles may borrow the monoterpenes from the pines they feed on, or alternatively synthezise them de novo. Again, compounds such as citral and citronellol are relatively nonspecic toxicants synthesized in the defensive secretions of ants or termites. Their odours may be sucient to deter an attacker, while the vapour may have an irritating eect on the predators skin (Harborne, 1993).

Iridoids
The iridoids are a group of bitter-tasting monoterpenoid lactones that have a restricted occurrence in dicotyledonous angiosperms. They are found in about 70 families belonging to some 13 orders (Jensen et al., 1975). Typical iridoid-containing plants are found in the Labiatae, Plantaginaceae, Scrophulariaceae and Valerianaceae. Iridoids are lactones, commonly with a glucose attachment to the hydroxyl of the lactone ring. A typical structure is loganin (Figure 4), which occurs in Strychnos nux-vomica fruit to the extent of 45% dry weight. Iridoid aglycones, after hydrolysis of the sugar, are highly unstable and usually disintegrate. A second group of iridoids have the ve-membered ring of carboxylic iridoids opened, giving rise to seco-iridoids, which have as a result an additional aldehyde function. The seco-iridoid derived from loganin is seco-loganin (Figure 4),
H HO H O CH3 H OGlc CO2Me HO2 C H2C CO2Me

O OGlc Seco-loganin OH H

Loganin

O H H CH3 O

O HOH2 C H

O OGlc

cis-trans-Nepetalactone
Figure 4 Structures of four iridoids.

Catalpol

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Terpenoids: Lower

a widespread substance in the Caprifoliaceae. Secoiridoids have another role in plant metabolism as biosynthetic precursors of terpene alkaloids. Thus secologanin can condense with the amino acid tryptophan to give rise to the alkaloid corynantheine in Corynanthe johimbe (Rubiaceae). A few volatile iridoids without glucose attachment are present in plants, a notable example being nepetalactone (Figure 4), the active principle of catmint Nepeta cataria. The plant has a peculiar attraction to members of the cat family. However, the purpose of its production in the plant is more likely to be related to its insect-repellent properties. Structures similar to nepetalactone occur in the defensive secretions of ants, stick insects and beetles (Harborne, 1993). Plants containing iridoids have been used in folk medicine in the treatment of inammation and as a bitter tonic. Valepotriates, iridoids in Valeriana, are sedative agents. Iridoids can be toxic and, in medieval times, bread contaminated with iridoid-containing Rhinanthus seed caused human deaths. Iridoids have been ingested from food plants by various buttery larvae and hence provide the adult with protection from bird predation. For example, catalpol from Plantago species is sequestered and stored in this way by Euphydryas butteries in North America (Harborne and Tomas-Barberan, 1991).

animals and are encountered in insect defence secretions. Some insect pheromones are sesquiterpenoid in nature. This applies to (E)-b-farnesene, an alarm pheromone of aphids. It is also relevant here that one class of insect hormone, juvenile hormone, is sesquiterpenoid in nature and such hormones (e.g. JH III) and hormone-mimics (e.g. juvabione) have been encountered in some quantity in certain plants (Harborne, 1993).

Abscisic Acid as a Plant Growth Substance

Abscisic acid (ABA) was rst discovered in 1964 as a dormancy factor in plants. It accumulates, for example, in dormant potato tubers and the dormant buds on trees. It has little to do with leaf abscission, as the name incorrectly hints, and it was only later that its regulatory role in stomatal opening was established. ABA is a sesquiterpene acid, with additional keto and hydroxyl functions. It occurs naturally as the optically active S-( 1 )-form (Figure 3). The unnatural R-( 2 )-ABA has been synthesized but this has no eect whatsoever on stomatal opening. Two conjugates of ABA are regularly found with it, the glucose ester and the 1-glucoside. There are several related metabolites known, including phaseic acid (Figure 6) formed from ABA during inactivation, and xanthoxin (Figure 3) which is a more potent growth inhibitor than ABA. ABA is present universally in owering plants. However, it is absent from liverworts, where its role may be taken by the stilbenoid lunularic acid. All organs of higher plants that have been analysed so far show the presence of this hormone, but the concentrations vary considerably, ranging from about 10 mg kg 2 1 in ripening fruit (e.g. avocado, rose hip) to about 10 mg kg 2 1 in water plants. The availability of mutant plants decient in ABA has provided new information on the eects of this key plant hormone. Mutants of tomato plants lacking ABA lose water faster than they can replace it because their stomata cannot close. They are permanently wilted. When ABA is supplied externally to these wilty mutants, their stomata close and they become turgid (Neil and Horgan, 1985). This, combined with many earlier results, conrms that the regulation of stomatal opening is the best-established function for ABA. The involvement of ABA in the control of plant dormancy is another established role. In the case of trees and shrubs, there is still some uncertainty, although the ABA content of buds falls as the intensity of dormancy decreases in the spring. It is clear that ABA has a well dened role in seed dormancy. Thus, seeds of maize mutants decient in ABA germinate prematurely on the cob. They are known as viviparous mutants and normal seed maturation can only be restored by adding ABA.

Sesquiterpenoids
The sesquiterpenoids are chemically dened by their formation from three isoprene units via the common C15 precursor farnesyl diphosphate (see Figure 3). They cooccur with monoterpenoids in plant essential oils and can usually be distinguished by their higher boiling points. There are three main groups, according to whether they are acyclic (e.g. farnesol), monocyclic (e.g. bisabolol) or bicyclic (e.g. b-cadinene) (Figure 5). Some are simple unsaturated hydrocarbons, but most have other functional groups as well. The derived sesquiterpenoid abscisic acid is a plant hormone and is discussed separately below. Sesquiterpenoids of one large group also have a lactone function, and these are also considered later. The main occurrence of sesquiterpenes is in plant essential oils, and some structures such as bisabolol, caryophyllene and b-cadinene are widely present in leaf oils of plants in the Labiatae, Rutaceae, Myrtaceae and Pinaceae. Other compounds are of more restricted occurrence. Carotol, for example, is characteristic of the carrot genus Daucus in the Umbelliferae. Another sesquiterpenoid, rishitin, is conned to the Solanaceae, and here it is only produced after a plant such as the potato is infected by microorganisms (Bailey and Manseld, 1982). Besides occurring in higher plants, sesquiterpenes are well represented in bryophytes (Asakawa, 1992) and in microorganisms. They are found additionally in marine
4

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Terpenoids: Lower

HO

CH3 H

-Bisabolol CH3

-Cadinene H

HO Carotol

-Caryophyllene

(E )--Farnesene CH2 OH Farnesol HO

CH3 H

HO

Juvabione
Figure 5 Structures of some plant sesquiterpenoids.

Rishitin

O O Phaseic acid

CO2 H

Hence, ABA is essential for seed maturation, stimulating the accumulation of seed proteins during the dormant phase (Milborrow and Netting, 1991). Current research in ABA is centred on its more general involvement in the adaptation of plants to abiotic environmental stresses. There is evidence for reversible protein phosphorylation and for modication of cytosolic calcium levels as intermediates in an ABA signal transduction cascade (Leung and Giraudat, 1998).

OH O

CO2 Glc

Sesquiterpene Lactones: Occurrence and Biological Properties

Sesquiterpene lactones are chemically distinct from other sesquiterpenoids by the presence of a g-lactone system.
5

ABA glucose ester

Figure 6 Structures of two abscisic acid metabolites.

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Terpenoids: Lower

Many have antitumour activity but their considerable cytotoxicity has so far prevented any useful anticancer applications. They have a variety of other biological properties, as will be described later. These lactones are classied biogenetically, according to the carbocyclic skeleton present, into four groups: germacranolides with a 10-membered ring, e.g. alatolide; eudesmanolides with two fused six-membered rings, e.g. alantolactone; guaianolides with fused ve- and sevenmembered rings and a methyl at C4, e.g. artabsin; and pseudoguaianolides, as guaianolides but with a methyl at C5 (compare artabsin and ambrosin, Figure 7). Besides these four main types, there are a variety of other lactones, formed by further modication of the carbon skeleton during biosynthesis. Germacranolides are recognized as the most primitive type and other skeletal classes can be derived biogenetically from them (Seaman, 1982). Unfortunately, very little biosynthetic study has been devoted to these lactones, so that we know very little about the exact intermediates involved. Other structural modications that can take place include hydroxylation, dimerization, glycosylation and the introduction of chlorine or aromatic substituents. At least 4000 lactones have been described and the majority of them have been obtained from a single plant family, the Compositae, where they are characteristic (Seaman, 1982). They have been reported occasionally in about 16 other angiosperm families, the only other major source being the Umbelliferae. Additionally, they have been found once in the gymnosperms, in Cupressaceae, and from a few fungi and from liverworts. In the Compositae, these lactones are found particularly in aerial parts, leaves and owering heads in concentrations of about 5% dry weight. They are often located in leaf trichomes or in surface wax. Occasionally, their occurrence extends to the roots, as in chicory, Cichorium intybus, where they are present in the latex. Complex mixtures are the rule rather than the exception and from 3 to 15 components may be found in a given plant tissue. The main roles assigned to sesquiterpene lactones today is as defensive agents against herbivory and microbial invasion. For example, 10-deoxylactucin and lactupicrin, the major lactones of chicory, Cichorium intybus, occur in sucient quantity to deter insect feeding and are bitter enough to protect the plant from mammalian browsing (Rees and Harborne, 1985). In general, lactones exhibit signicant antifeedant properties against locusts and army worms and reduce the survival of insect larvae and adults. Certain lactones (e.g. geigerin) are toxic to livestock and others (e.g. parthenin) are well known to cause allergic contact dermatitis (as in Parthenium hysterophorus). Additionally, many sesquiterpene lactones have been shown to have both antibacterial and antifungal activity. Parthenolide, which occurs in feverfew, Tanacetum parthenium, is recognized as the antifungal principle of this plant
6

CH2 OH O O H CH2 OH H H CH2 O Alatolide (germacranolide)

CH3 CH3 CH3 H O O H CH2

CH3

Alantolactone (eudesmanolide)

HO

CH3

H H H H

CH3

H CH3 CH3 H O Artabsin (guaianolide) O

H CH3 O O Ambrosin (pseudoguaianolide) CH2

CH3

H H O CH2 O

CH3 H

H H CH2 OH H O

O CH3 HO H H O H CH3

8-Deoxylactucin (guaianolide)

Geigerin (guaianolide)

H HO H

CH3

CH3

CH3 H O CH2 O H H O O Parthenolide (germacranolide) H CH3

CH3 O

Parthenin (pseudoguaianolide)

Figure 7 Structure of representative sesquiterpene lactones.

(Blakeman and Atkinson, 1979). The same compound, incidentally, is responsible for the antimigraine properties of feverfew, one of the few lactone-containing members of the Compositae useful in medicine.

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Terpenoids: Lower

References
Asakawa Y (1992) Chemical constituents of the Hepaticae. Fortschritte der Chemie Organischer Naturstae 42: 1286. Bailey JA and Manseld JW (eds) (1982) Phytoalexins. Glasgow: Blackie. Banthorpe DV, Brench SA, Njar VCO, Osborne MG and Watson DG (1986) Ability of plant callus cultures to synthesise and accumulate lower terpenoids. Phytochemistry 25: 629636. Blakeman JP and Atkinson P (1979) Antimicrobial properties and possible role in hostpathogen interactions of parthenolide, a sesquiterpene lactone isolated from the glands of Chrysanthemum parthenium. Physiological Plant Pathology 15: 183192. Charlwood BV and Banthorpe DV (eds) (1991) Methods in Plant Biochemistry, vol. 7, Terpenoids. London: Academic Press. Charlwood BV, Charlwood KA and Molina-Torres J (1990) Accumulation of secondary compounds by organised plant cultures. Proceedings of the Phytochemical Society of Europe 30: 167200. Harborne JB (1993) Introduction to Ecological Biochemistry. London: Academic Press. Harborne JB and Tomas-Barberan FA (eds) (1991) Ecological Chemistry and Biochemistry of Plant Terpenoids. Oxford: Clarendon Press. Jensen SR, Nielsen BJ and Dahlgren R (1975) Iridoid compounds, their occurrence and systematic importance in angiosperms. Botaniska Notiser 128: 148180. Leung J and Giraudat J (1998) ABA signal transduction. Annual Review of Plant Physiology, Plant Molecular Biology 49: 199222. Lichtenthaler HK (1999) The 1-deoxy-d -xylulose pathway of isoprenoid biosynthesis in plants. Annual Review of Plant Physiology, Plant Molecular Biology 50: 4766. Milborrow BV and Netting AG (1991) Abscisic acid and derivatives. In: Charlwood BV and Netting AG (eds) Methods in Plant Biochemistry, vol 7, Terpenoids, pp. 213262. London: Academic Press. Neil SJ and Horgan R (1985) ABA production and water relations in wilty tomato mutants subjected to water deciency. Journal of Experimental Botany 36: 12221231.

Parry AD (1993) Abscisic acid metabolism. In: PJ Lee (ed.) Methods in Plant Biochemistry, vol 9, Enzymes of Secondary Metabolism, pp. 381 402. London: Academic Press. Rees SB and Harborne JB (1985) The role of sesquiterpene lactones and phenolics in the chemical defence of the chicory plant. Phytochemistry 24: 22252231. Seaman FC (1982) Sesquiterpene lactones as taxonomic characters in the Asteraceae. The Botanical Review 48: 121595.

Further Reading
Charlwood BV and Banthorpe DV (eds) (1991) Methods in Plant Biochemistry, vol. 7, Terpenoids. London: Academic Press. Harborne JB (1993) Introduction to Ecological Biochemistry. London: Academic Press. Harborne JB and Tomas-Barberan FA (eds) (1991) Ecological Chemistry and Biochemistry of Plant Terpenoids. Oxford: Clarendon Press. Jensen SR, Nielsen BJ and Dahlgren R (1975) Iridoid compounds, their occurrence and systematic importance in angiosperms. Botaniska Notiser 128: 148180. Leung J and Giraudat J (1998) ABA signal transduction. Annual Review of Plant Physiology, Plant Molecular Biology 49: 199222. Lichtenthaler HK (1999) The 1-deoxy-d -xylulose pathway of isoprenoid biosynthesis in plants. Annual Review of Plant Physiology, Plant Molecular Biology 50: 4766. Milborrow BV and Netting AG (1991) Abscisic acid and derivatives. In: Charlwood BV and Netting AG (eds) Methods in Plant Biochemistry, vol 7, Terpenoids, pp. 213262. London: Academic Press. Parry AD (1993) Abscisic acid metabolism. In: PJ Lea (ed.) Methods in Plant Biochemistry, vol 9, Enzymes of Secondary Metabolism, pp. 381 402. London: Academic Press. Seaman FC (1982) Sesquiterpene lactones as taxonomic characters in the Asteraceae. The Botanical Review 48: 121595.

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