Genetic component of variation in chemical defense ofOreina gloriosa (Coleoptera: Chrysomelidae)

Defensive secretions of adultOreina gloriosa, liberated at the surface of the pronotum and elytra, contain a complex mixture of cardenolides, and ethanolamine. Proportions and concentrations of constituents determined by reverse-phase HPLC show considerable variation among individual beetles. Heritabilities of proportions of five main components were estimated by mother-offspring regression providing a validation of the less reliable full-sib correlation estimates. Average heritabilities based on the two methods were 0.51 and 0.58, respectively, estimated by using offspring of two age groups. Regression estimates of 2- and 10-week-old offspring differed significantly for one secretion constituent (RT16). Heritability estimates of concentrations of 16 secretion components were calculated by full-sib correlation analysis. Average heritability was 0.45, indicating a significant genetic component. Estimates did not differ significantly between the two age groups. We also estimated heritabilities of concentrations by a two-way model including data from offspring of both age groups. Heritability estimates based on this model are thought to correspond approximately to estimates based on samples from natural populations. The average of these estimates was lower (h ² =0.31) than the average heritability of each age group separately (h ² =0.45), suggesting a developmental effect on variation in chemical defense ofO. gloriosa.     

Production of cardenolides versus sequestration of pyrrolizidine alkaloids in larvae ofOreina species (Coleoptera,Chrysomelidae)

Adult leaf beetles of the genusOreina are known to be defended either by autogenously produced cardenolides or by pyrrolizidine alkaloids (PAs) sequestered from the food plant, or both. In this paper we analyze larvae of differentOreina species and show that the larvae contain the same defensive toxins as the adults in quantities similar to those released in the adults' secretion. Both classes of toxins are found in the body and hemolymph of the larvae, despite their different origins and later distribution in the adults. Larvae of sequestering species differed in their PA patterns, even though they fed on the same food plants. The concentration in first-instar larvae of a PA-sequestering species was similar to that in fourth-instar larvae. In all stages examined, the amount of PAs per larva did not greatly exceed the estimated uptake of one day. Eggs of two oviparous species contained large concentrations of the adult's toxins, while neonates of a sequestering larviparous species had no PAs.     

Distribution of autogenous and host-derived chemical defenses inOreina leaf beetles (Coleoptera: Chrysomelidae)

The pronotal and elytral defensive secretions of 10Oreina species were analyzed. Species feeding on Apiaceae, i.e.,O. frigida andO. viridis, or on Cardueae (Asteraceae), i.e.,O. bidentata, O. coerulea, andO. virgulata, produce species-specific complex mixtures of autogenous cardenolides.O. melanocephala, which feeds onDoronicum clusii (Senecioneae, Asteraceae), devoid of pyrrolizidine alkaloids (PAs) in its leaves, secretes, at best, traces of cardenolides. Sequestration of host-plant PAs was observed in all the other species when feeding on Senecioneae containing these alkaloids in their leaves.O. cacaliae is the only species that secretes host-derived PA N-oxides and no autogenous cardenolides. Differences were observed in the secretions of specimens collected in various localities, because of local differences in the vegetation. The other species, such asO. elongata, O. intricata, andO. speciosissima, have a mixed defensive strategy and are able both to synthesize de novo cardenolides and to sequester plant PA N-oxides. This allows a great flexibility in defense, especially inO. elongata andO. speciosissima, which feed on both PA and non-PA plants. Populations of these species were found exclusively producing cardenolides, or exclusively sequestering PA N-oxides, or still doing both, depending on the local availability of food-plants. Differences were observed between species in their ability to sequester different plant PA N-oxides and to transform them. Therefore sympatric species demonstrate differences in the composition of their host-derived secretions, also resulting from differences in host-plant preference. Finally, within-population individual differences were observed because of local plant heterogeneity in PAs. To some extent these intrapopulation variations in chemical defense are tempered by mixing diet and by the long-term storage of PA N-oxides in the insect body that are used to refill the defensive glands.     

Composition of larval secretion ofChrysomela lapponica (Coleoptera,Chrysomelidae) and its dependence on host plant

The defensive secretion ofChrysomela lapponica larvae, which is produced by nine pairs of exocrine dorsal glands, has been chemically analyzed. TheC. lapponica larvae were kept in the laboratory on leaves of either birch (Betula pendula), alder (Alnus glutinosa), or willow (Salix fragilis). Larvae developed normally on birch and willow, whereas those on alder died within a few days. GC-MS analyses of the secretion of larvae on birch and willow revealed that the composition of this secretion differs distinctly from the known ones of several otherChrysomela species feeding exclusively on Salicaceae. In the exocrine secretion of larvae on birch, 69 compounds were identified, which included the main components isobutyric acid, 2-methylbutyric acid, and esters of the two. Several of the esters have not been reported previously from nature. The alcoholic components of the esters may be hydrolysis products ofBetula glycosides. Most components of the secretion of larvae feeding on birch were also found in the secretion of larvae feeding on willow. In addition, major amounts of benzoic acid and salicylalcohol were present in the secretion of the larvae feeding on willow.C. lapponica obviously acquires salicylalcohol by hydrolysis of salicin from willow leaves. However, in contrast to otherChrysomela species,C. lapponica larvae oxidize only traces of salicylalcohol to salicylaldehyde. The repellent activity of single authentic compounds of the secretion of larvae feeding on birch and willow, respectively, was tested in laboratory bioassays with ants (Myrmica sabuleti). Biosynthetic pathways to some identified compounds are suggested and discussed under evolutionary and functional aspects.     

Trirhabda canadensis (Coleoptera: Chrysomelidae) responses to plant odors

The responses of the goldenrod leaf beetleTrirhabda canadensis to host and nonhost volatile odors were tested in a Y-tube olfactometer in the laboratory. Beetles preferred host to nonhost odors and were sensitive to concentrations of host odor. Beetles distinguished between host and nonhost volatiles of only one of the two nonhostSolidago species; host volatiles were preferred to all nonhost volatiles at the family and order levels. In other words, all nonhosts above the genus level had similar effects on beetle responses. Although the odors of most nonhosts were neutral (i.e., neither attractive nor repellent) to the beetles as tested against air, this neutrality disappeared if the odors of two or more nonhosts were added to the host odor and beetles were given a choice between this mixture and pure host odor. Given this choice, they strongly preferred pure host odor, which suggests that diversity of odors per se is unattractive to the beetles. Beetles walked rather than flew to locate their hosts in the field, and their movements suggest that they used olfactory cues to locate hosts.     

Distribution of adult defense glands in chrysomelids (Coleoptera: Chrysomelidae) and its significance in the evolution of defense mechanisms within the family

Defense glands were examined in the adults of 65 species belonging to 10 different subfamilies. They were found in the pronota and elytra of members of the subfamilies Criocerinae, Chrysomelinae, Galerucinae, and Alticinae. It is suggested that these glands appeared monophyletically in the course of evolution and that the absence of glands in several species of the two most evolved subfamilies is a secondary event, explained by the presence of alternative efficient defensive behaviors: reflex bleeding in the Galerucinae and escape mechanism of jumping in the flea beetles. It is also suggested that a large distribution of the glands at the surface of the beetles is a primitive condition and that in the course of evolution only the glands most efficiently located along the edges of the pronotum and elytra were maintained. Such evolution has occurred several times. Alternative and complementary defensive mechanisms are also listed and discussed.     

Evidence of an aggregation pheromone in the flea beetle,Phyllotreta Cruciferae (Goeze) (Coleoptera: Chrysomelidae)

Laboratory olfactometer bioassays and field trapping experiments showed that the flea beetle,Phyllotreta cruciferae (Goeze), was highly attracted by oilseed rape(Brassica napus L.) when flea beetles were on the plant. This attraction was mediated by a flea beetle-produced aggregation pheromone based upon: (1) Oilseed rape damaged mechanically, or byP. cruciferae, or by diamondback moth,Plutella xylostella (L.), did not attractP. cruciferae. (2) Contact with the plants or feeding was required for the production of aggregation pheromone because oilseed rape alone was not attractive when separated from flea beetles by a screen. (3) Equal numbers of males and females were attracted.     

Odors influence choice of oviposition sites byDiabrotica virgifera virgifera (Coleoptera: Chrysomelidae)

FemaleDiabrotica virgifera virgifera LeConte were allowed to choose between oviposition substrates that were and those that were not associated with potential sources of semiochemicals. Females deposited over five times more eggs on moist towelettes that were treated with homogenates of female abdomens than on towelettes treated with distilled water. Similar results were obtained when screening separated the homogenates from the towelettes, indicating that odors alone could elicit the response. In contrast, females did not choose towelettes that had previously been used for oviposition or towelettes containing eggs over unused towelettes. Further tests with homogenates of abdomens and a bacteriostatic agent (sorbate) indicated that the females were probably responding to bacterial odors rather than an oviposition-enhancing pheromone. Four strains of bacteria were isolated from a homogenate of female abdomens; females deposited 4 to 16 times more eggs on substrates with odors of the bacteria than on substrates with odors of uninoculated nutrient agar. In no-choice tests, bacterial odors did not increase the number of eggs deposited per female beetle; however, in choice tests with dishes that tended to retain any beetles that entered, there were more eggs per female (but not more beetles) after 24 hr in dishes with bacterial odors than in those without the odors. Females also chose dishes with odors of excised maize (Zea mays L.) roots or elevated levels of carbon dioxide over control dishes.Mention of a commercial or proprietary product does not constitute an endorsement for its use by USDA.     

Economics of chemical defense in chrysomelinae

Chemical defense in chrysomelid larvae (subtribe Chrysomelina and Phyllodectina) is reviewed. Most species secrete autogenous monoterpenes. The diversity of their secretion is interpreted as a mechanism to reduce adaptation by predacious arthropods. The consequences of a host plant shift to the Salicacae are explored. Salicin from these host plants is used as a precursor for the salicylaldehyde secreted by the larvae of many species. This offers several advantages. It provides the larvae with an inexpensive and efficient defense. The recovery of the glucose moiety of the salicin contributes significantly to the larval energy budget. Adults sequester salicin in the eggs at concentrations which are toxic to ants. Owing to this maternal provisioning, neonate larvae produce salicylaldehyde from hatching onwards, whereas other species secreting monoterpenes are not protected at hatching. The secretion of salicylaldehyde by different species is considered to be chemical mimicry reinforcing visual aposematic signals.     

Coevolutionary adaptations of rootworm beetles (Coleoptera: Chrysomelidae) to cucurbitacins

The cucurbitacins are oxygenated tetracyclic triterpenoids produced as secondary plant compounds by nearly all genera of Cucurbitaceae. The very bitter and toxic cucurbitacins are effective semiochemicals acting ecologically as allomones to protect the Cucurbitaceae from attack by a variety of invertebrate and vertebrate herbivores. For the Luperini (Coleoptera: Chrysomelidae: Galerucinae) the cucurbitacins have become kairomones for host selection, affecting the behavior of this large group of 1500 species of Aulacophorina (Old World) and Diabroticina (New World) by arrest and compulsive feeding. When feeding on bitter cucurbits these beetles sequester large amounts of cucurbitacins in their blood and tissues, and these act as allomones to deter predation. Specific detoxification and excretory mechanisms of the Diabroticina enable these beetles to avoid the toxic effects of the cucurbitacins.     

Electroantennogram responses ofTrirhabda bacharides (weber) (Coleoptera: Chrysomelidae) to plant volatiles

Electroantennograms (EAGs) were recorded fromTrirhabda bacharides Le Conte (Coleoptera: Chrysomelidae) females to 28 odorants and volatiles emanating from leaves of 26 plant species including three known host plants,Baccharis halimifolia L.,B. neglecta Britt, andB. salicina T. & G. (Compositae: Asterae). Antennal receptors were highly responsive to components of the green leaf volatile complex, especially 6-carbon saturated and monounsaturated alcohols andtrans-2-hexenal. EAGs elicited by heptanal were greater than those elicited by any other saturated aldehyde. Oxygenated monoterpenes were more active than monoterpene hydrocarbons. -Bisabolol was the most active sesquiterpene. In general, EAGs to volatiles emanating fromBaccharis and other composite species were greater than those elicited by species from other families. Furthermore, when one considers only plants occurring in the environs ofT bacharides hostB. neglecta, EAGs elicited by its host plant were at least two times greater than those elicited by other coinhabiting plants.Mention of a trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products or vendors that may also be suitable.     

Anthraquinones in different developmental stages ofGaleruca tanaceti (Coleoptera,Chrysomelidae)

The overwintering eggs and the larvae of the leaf beetleGaleruca tanaceti (L.) contain hydroxylated anthraquinones. In both developmental stages, l,8-dihydroxy-3-methylanthraquinone (= chrysophanol) and 1,8-di-hydroxyanthraquinone (= chrysazin) were detected by GC-MS and GC-FTIR analyses. In the eggs, chrysazin was found only in traces. Anthraquinones were also present in ovaries and hemolymph of gravid females, which were investigated in order to examine the incorporation of these substances into the eggs. Neither in acidified nor in nonacidified extracts of the host plantsTanacetum vulgäre L. andAchillea millefolium L. were anthraquinones found. The activity of these anthraquinones as chemical defense substances was proved in bioassays with the antMyrmica ruginodis NYL. Further possible biological significances of anthraquinones are discussed.     

Impact of acidic deposition onEncelia farinosa gray (Compositae: Asteraceae) and feeding preferences ofTrirhabda geminata horn (Coleoptera: Chrysomelidae)

Container grownEncelia farinosa were exposed to three 3-hr episodes of acidic fog (pH 2.5) typical of events in southern California. Adults and larvae of the specialist leaf-feeding herbivore,Trirhabda geminata, preferred to feed on the acidic-treated foliage compared to control fogged (pH 6.3–6.5) foliage. Previous feeding damage on the plants did not affect feeding preference. The acidic-fogged foliage was significantly higher in total nitrogen and soluble protein but not different from control-treated tissue in water content. Stress on native populations of this drought-deciduous shrub caused by atmospheric pollutants may also result in altered feeding ecology of the beetle.     

Effect of allyl isothiocyanate on field behavior of crucifer-feeding flea beetles (Coleoptera: Chrysomelidae)

When water traps baited with allyl isothiocyanate (AIC)diffusing through polyvinyl chloride (PVC) and rubber membranes were used to monitor four species of crucifer-feeding flea beetle adults in a rutabaga field at L'Assomption, Que. in 1980–1981, differential responses to AIC were observed.Phyllotreta cruciferae was more attracted to AIC thanP. striolata, whereas the behavior ofPsylliodes punctulata was not affected by the presence of AIC. The traps with the PVC membrane caught significantly more flea beetles than the traps with the rubber membrane in 1980, but caught a similar number in 1981. Sticky traps covered with AIC mixed with Tangletrap® caught significantly more flea beetles than control sticky traps.     

Defensive adaptations of eggs and adults ofGastrophysa cyanea (Coleoptera: Chrysomelidae)

Egg clusters and adults ofGastrophysa cyanea are conspicuous and, like their larvae, are chemically protected. The eggs owe their bright yellow color primarily to -carotene and, in addition, contain substantial quantities of oleic acid. At natural concentrations oleic acid effectively deters many species of ants from feeding. The use of fatty acids as deterrents against ants is discussed as a possible widespread phenomenon among insects. During defensive confrontations, adults ofG. cyanea exhibit avoidance behavior and may also feign death. In addition, the adults may autohemmorhage or secrete a fluid from elytral or pronotal pores in response to traumatic stimuli. The secretions are effective against ants and contain a mixture of hydrocarbons as well as terpenoid components. The pattern of ontogenetic modification in the defensive chemical repertoire ofG. cyanea is discussed.     

Attraction of adultDiabrotica (Coleoptera: Chrysomelidae) to corn silks and analysis of the host-finding response

Volatile chemicals from corn silks attractedDiabrotica virgifera virgifera LeConte andD. barberi (Smith and Lawrence). The behavioral response of both species of beetles to the host plant was typified by four distinct phases: perception, random movement, orientation to the source, and search with reorientation. The perception phase was composed of stationary behaviors, while the random, orientation, and search phases were composed of directed and nondirected movements. Each of the movement phases had a characteristic response pattern composed of the ratio of upwind, lateral, and downwind walking and flight movements, which affected net displacement of the beetle in the flight tunnel. The perception phase occurred within and between the other phases and was responsible for initiating changes from one movement phase to another (based on the presence or absence of volatiles from corn silks). Host finding was flexible, and the response pattern fit a flow-chart type of response, rather than a single stereotyped sequence of behaviors.     

Identification of female-produced sex pheromone from banded cucumber beetle,Diabrotica balteata leconte (Coleoptera: Chrysomelidae)

A sex pheromone produced by female banded cucumber beetle adults,Diabrotica balteata LeConte, was isolated from volatiles trapped on Porapak Q and identified as 6,12-dimethylpentadecan-2-one. The structure was elucidated by spectroscopic analyses and confirmed by synthesis. The synthesized racemic compound was equal to the purified natural pheromone in eliciting responses by banded cucumber beetle males to field traps. A doseresponse characteristic was demonstrated for the racemic material formulated on filter paper or rubber septa and placed in field traps. The absolute configuration at the C-6 and C-12 positions was not established.This article reports the results of research only. Mention of a proprietary product does not constitute an endorsement or the recommendation for its use by USDA.This research was initiated by Dr. P.L. Guss, deceased, Research Chemist, Northern Grain Insects Research Laboratory, ARS, USDA, Brookings, South Dakota. Dr. Guss conducted the preliminary purifications and bioassays that led to the isolation and identification of this pheromone.     

Use of chemical variation and predation as plant defenses byEncelia farinosa against a specialist herbivore

Larvae of the monophagous herbivore,Trirhabda geminata, selectively eat particular plants and plant parts of its natural host,Encelia farinosa. Measurements of leaf damage and larval positions on branches through time support this observation. Time-lapse movie photography revealed that larvae are sufficiently mobile to search most of a plant in a 48-hr period and that aggregations were the result of larval activity and not directly the result of oviposition. Experiments withT. geminata larvae on artificial diets containing a range of natural concentrations of chemical extracts fromE. farinosa leaves showed that the larvae grew significantly slower and had a lower overall survivorship at the high concentration. Combining the results of all choice tests, larvae appeared unable to distinguish between high- and low-concentration agar diets. Considered individually, larval preferences for natural production concentrations changed as the season progressed. Early-season larvae preferred low-concentration leaves, while late-season larvae preferred high-concentrations. Measurements of chemical and nitrogen content of leaves selected by larvae in the field confirmed this pattern. Percent parasitism in field-collected larvae increased with season as the larval population decreased. This combination of slowed growth and increasing parasitism and predation is a putative defense strategy ofEncelia farinosa to prevent adaptation by a specialist herbivore to the total range of compounds elaborated.     

Evidence for chemical defense in tropical green algaCaulerpa ashmeadii (Caulerpaceae: Chlorophyta): Isolation of new bioactive sesquiterpenoids

Results of field feeding preference studies with 12 species of tropical green algae of the genusCaulerpa showed thatC. ashmeadii was preferred least by herbivorous fishes. Chemical investigations ofC. ashmeadii demonstrated the presence of high concentrations of sesquiterpenoid metabolites. The chemical isolation and structural elucidation of five majorC. ashmeadii metabolites, as well as the results of field feeding preference, antimicrobial, and ichthyotoxicity assays demonstrating the biological activities of these metabolites are reported here.     

Inter- and intrapopulation variation of the pheromone,ipsdienol produced by male pine engravers,Ips pini (Say) (Coleoptera: Scolytidae)

We determined the chirality of ipsdienol in individual male pine engravers,Ips pini (Say), from New York, California, and two localities in British Columbia (BC). Both quantity and chirality of ipsdienol varied significantly between and within populations ofI. pini. Beetles from California and southeastern BC produced primarily (R)-(–)-ipsdienol with mean ratios of (S)-(+) : (R)-(–) of 9 : 91 and 11 : 89, respectively, while beetles from New York produced primarily (S)-(+)-ipsdienol with a mean (S)-(+) : (R)-(–) ratio of 57 : 43. A population from southwestern BC was unlike any other known western population, producing primarily (S)-(+)-ipsdienol with a mean (S)-(+) : (R)-(–) ratio of 66 : 34. In contrast to the unimodal chirality profiles for ipsdienol production in populations from California and southeastern BC, the profiles of the populations from southwestern BC and New York were bimodal, with a common mode at approximately 44 : 56 (S)-(+) : (R)-(–). Bimodality in the profiles of ipsdienol chirality in two populations ofI. pini and remarkably high levels of intrapopulation variation in pheromone chirality in all four populations suggest that evolutionary change in pheromone channels of communication could occur, possibly in response to artificial selection pressures such as mass trapping.