Chemosensory basis of behavioural plasticity in response to deterrent plant chemicals in the larva of the Small Cabbage White butterfly Pieris rapae

Behavioural and electrophysiological responsiveness to three chemically different secondary plant substances was studied in larvae of Pieris rapae L. (Lepidoptera: Pieridae). Three groups of caterpillars were studied that during their larval development were exposed to different rearing diets: an artificial diet or one of two host-plants, cabbage, Brassica oleracea, or nasturtium, Tropaeolum majus. In dual-choice leaf disc assays, caterpillars reared on cabbage were strongly deterred by the phenolic chlorogenic acid, the flavonol glycoside naringin and the alkaloid strychnine. However, behavioural plasticity was found in caterpillars reared on nasturtium or artificial diet in that these did not discriminate against chlorogenic acid. Caterpillars reared on the artificial diet were also significantly less sensitive to naringin and strychnine in the behavioural assay. Electrophysiological studies of the maxillary sensilla styloconica revealed that the deterrent neuron in the medial sensillum, but not in the lateral sensillum, of cabbage-reared caterpillars was more sensitive than the same neuron type of caterpillars reared on nasturtium or artificial diet. We conclude that (1) the diet-induced behavioural habituation to deterrents can at least partly be explained by chemosensory desensitisation of a generalist type of maxillary deterrent neuron; (2) behavioural cross-habituation to the three structurally diverse deterrent compounds can be traced back to cross-sensitivity for these compounds in the same gustatory neuron.     

Effects of plant nutrition on the balance of insect relevant cardenolides and glucosinolates in Erysimum cheiranthoides

The possible effects of environmental stress on plant chemistry that are important to herbivorous insects were examined by growing a wild crucifer, Erysimum cheiranthoides, under different nutrient regimes. Oviposition by the cabbage butterfly, Pieris rapae, is thought to be affected by the balance of glucosinolates (stimulants) and cardenolides (deterrents) at the surface of leaves. E. cheiranthoides seedlings were provided with three levels of nitrogen and two levels of sulfur for a period of 15 days before analysis of semiochemicals in whole leaf tissue and at the surface of the foliage. The ratio of cardenolides to glucosinolates in the plants at elevated C/N ratios followed the carbon/nutrient balance hypothesis. However, a high nitrogen supply enhanced biomass production to the extent that concentrations of secondary compounds were unchanged or reduced. The concentration of glucosinolates (glucoiberin and glucocheirolin) at the surface was positively related to whole tissue levels. However, cardenolide (erysimoside and erychroside) concentrations, which were highest in leaf tissue of nitrogen-deficient plants, had the lowest surface levels on foliage of these plants. Possible reasons for differential expression of cardenolides and glucosinolates in a plant as a result of nutrient deficiency are discussed.     

Tarsal contact chemoreceptor response to glucosinolates and cardenolides mediating oviposition in Pieris rape

Abstract. Water-soluble extracts of a host crucifer (Brassica oleracea L.) and non-host crucifer (Erysimum cheiranthoides L.) and isolated pure cardenolides and glucosinolates were tested on Pieris rapae L. (Lepidoptera: Pieridae) butterflies in oviposition assays and by electrophysiological recordings from the contact-chemoreceptor sensilla of the prothoracic tarsi. Ten different glucosinolates stimulated oviposition to varying degree when put on non-host plant leaves. The most active compounds were glucobrassiein and gluconasturtiin (methylindole and phenylethyl aglycone), whereas glucocapparin, sinalbin. glucotropaeolin, sinigrin and glucoalyssinin had intermediate activity. Glucocheirolin, glucoerucin and glucoiberin (sulphur-containing aglycone) were significantly less active. Of eight cardenolides applied to host-plant leaves (100 μg each), four glycosides deterred oviposition strongly (erysimoside, erychroside, cymarin and K-strophanthin-β). Erycordin, helveticoside, digitoxin and strophanthidin had little or no deterrent activity. Sensilla located laterally on the prothoracic tarsi of female butterflies contained one receptor cell sensitive to sucrose. None of the tested extracts or pure compounds stimulated any cell in these sensilla. In contrast, the cells in the medial sensilla showed little or no sensitivity to sucrose. One cell was found to be sensitive to cardenolide glycosides. The threshold for one of the most active compounds, erychroside, was about 0.1 μg/ml (1.5 times 10^-7 M). Two receptor cells, characterized by spikes of differing amplitude, were sensitive to glucosinolates. One of these, with larger amplitude spikes, seemed to be the same as the cell sensitive to cardenolides. The threshold for the most active glucosinolates, glucobrassicin and gluconasturtiin was estimated to be below 0.1 mg/ml (2 times 10^-4 M). The neural activity of both classes of compounds, cardenolides and glucosinolates, was significantly correlated with their behavioural activity as deterrents or stimulants.     

Continuous exposure to the deterrents cis‐jasmone and methyl jasmonate does not alter the behavioural responses of Frankliniella occidentalis

Behavioural responses of Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), a generalist, cell sap‐feeding insect species with piercing‐sucking mouthparts, after continuous exposure to two deterrent secondary plant compounds are investigated. We compared in choice assays on bean leaf discs, the settling, feeding, and oviposition preferences of F. occidentalis females that had no experience with the two fatty acid derivatives methyl jasmonate and cis‐jasmone before testing (naïve thrips) vs. females that had been exposed to the deterrent compounds before testing (experienced thrips). The thrips were exposed to the deterrents at low or high concentrations for varied time periods and subsequently tested on bean leaf discs treated with the respective deterrent at either a low or a high concentration. Frankliniella occidentalis females avoided settling on the deterrent‐treated bean leaf discs for an observation period of 6 h, independent of their previous experience. Our results demonstrate that feeding and oviposition deterrence of the jasmonates to the thrips were not altered by continuous exposure of the thrips to the jasmonates. Habituation was not induced, neither by exposure to the low concentration of the deterrents nor by exposure to the high concentration. These results indicate that the risk of habituation to two volatile deterrent compounds after repeated exposure is not evident in F. occidentalis. This makes the two compounds potential candidates to be integrated in pest management strategies.     

Flavonoids from cabbage are feeding stimulants for diamondback moth larvae additional to glucosinolates: Chemoreception and behaviour

In caterpillars two styloconic contact chemoreceptors on the maxillary galea are assumed to contain the main taste receptors involved in host plant selection. The diamondback moth, Plutella xylostella L. is a specialist feeder of plants in the Brassicaceae, a plant family characterized by the biosynthesis of glucosinolates. We used pea (Pisum sativum L., Leguminosae) as a neutral non-host for a dual-choice leaf disc assay to quantify feeding stimulation by glucosinolates and flavonoids. Increasing concentrations of sinigrin resulted in significant preferences for sinigrin-treated leaf discs, with a threshold between 1 and 3 M. Millimolar concentrations of four of the five flavonol triglucosides likewise elicited a significant preference for flavonoid-treated leaf discs. A mixture of four flavonoids and sinigrin was significantly preferred over sinigrin-treated leaf discs alone. Vigorous unicellular electrophysiological responses of medial maxillary styloconic taste sensilla were observed in response to five glucosinolates (glucocapparin, sinigrin, glucobrassicin, glucoiberin, and gluconasturtiin). This medial taste neuron responded in a dose-dependent manner to a concentration series of sinigrin, with a threshold of response of ca. 1 M. The lateral sensillum styloconicum contained a neuron sensitive to sucrose, glucose, and fructose. However, no responses in the two types of maxillary styloconic sensilla to the phagostimulatory flavonoids could be detected, suggesting that other taste organs mediate chemoreception of flavonoids. We conclude that diamondback moth larvae employ a combination of biosynthetically distinct categories of feeding stimulants which allows for a higher degree of discriminatory ability than when this would be based on glucosinolates alone.     

Experience-based food consumption by larvae of Pieris rapae: addiction to glucosinolates?

When newly hatched larvae of P. rapae were transferred to cowpea foliage, they readily accepted this non-host as food, whereas later instars that had fed on cabbage rejected cowpea. However, when cowpea leaf discs were treated with aqueous extracts of cabbage foliage, they were accepted by cabbage-reared larvae. Experiments were conducted to determine whether larvae reared on one host plant would be stimulated to feed by extracts of other hosts. Larvae reared on Brassica juncea, Cleome spinosa, Tropaeolum majus, Sinapis alba, Alliaria petiolata, Barbarea vulgaris and cabbage (Brassica oleracea) were offered extracts of each of the other host plants on cowpea discs in choice assays. Larvae were generally stimulated to feed by extracts of all the alternate hosts, but quantitative differences in consumption occurred. In most cases, levels of discrimination between treatment and control cowpea discs showed no significant preference for extracts of the previously experienced plant. Since the test plants (and their extracts) contain glucosinolates of widely different structures, a general addiction to glucosinolates was suggested. A single glucosinolate, sinigrin, was sufficient to elicit feeding by cabbage-reared larvae. The time required for individual neonates to become addicted to glucosinolates as they fed on cabbage, as measured by refusal of cowpea, varied from 6 to 30 hours. Bioassays of cowpea extracts failed to show any deterrent activity and, therefore, supported the conclusion that addiction to glucosinolates is responsible for the fixation of P. rapae larvae on their host plants.     

Identification of glucosinolates on the leaf surface of plants from the Cruciferae and other closely related species

Leaf-surface extracts prepared from 18 non-cultivated (wild) plant species, derived from the Capparidaceae, Cruciferae, Resedaceae and Tropaeolaceae were ranked for their ability to stimulate oviposition by the cabbage root fly, and analysed for glucosinolates. A total of 28 different glucosinolates were identified. A clear relationship was detected between the indolyl-, benzyl- and the total glucosinolate composition on the leaf surface and oviposition preference by cabbage root fly females. However, as the results are not fully explained by differences in leaf surface glucosinolates, other important oviposition deterrents and stimuli on the leaf surface of these wild crucifers must also be present.     

菜粉蝶与其寄主植物金莲花

观察了用金莲花(TropaeolummajusL.)饲养的菜粉蝶PierisrapaeL.的发育情况,表明金莲花能使菜粉蝶完成世代发育,是其理想的寄主植物。国外有研究表明:将金莲花上取食的幼虫人为地转移到十字花科或其它寄主植物上,幼虫可正常取食,并最后发育为成虫;反之,将十字花科植物上取食的2龄至5龄的幼虫转移到金莲花上,则拒食外迁,最后全都饿死。然而,若将小麦胚芽人工饲料上饲养的幼虫转移到金莲花上,则幼虫正常取食。这是由于菜粉蝶幼虫在十字花科植物上取食的1龄阶段发展了对金莲花植株内所含的取食抑制化合物的敏感性,而在小麦胚芽人工饲料上取食的幼虫因一直接触取食抑制化合物而不对金莲花所含的取食抑制化合物产生敏感性之故。化学提取和昆虫行为实验证明,在金莲花中所含的取食抑制化合物的主要成分是绿原酸。     

The effect of modifying the glucosinolate content of leaves of oilseed rape (Brassica napus ssp. oleifera) on its interaction with specialist and generalist pests

Twenty eight Brassica napus lines were developed which had contrasting leaf glucosinolate profiles to those found in commercial oilseed rape cultivars. The lines varied both in the total amount of aliphatic glucosinolates and in the ratio of different side chain structures. The lines were used in field experiments to assess the manner by which glucosinolates mediate the interactions between Brasssica and specialist pests (Psylliodes chrysocephala and Pieris rapae) and generalist pests (pigeons and slugs). Increases in the level of glucosinolates resulted in greater damage by adult flea beetles (P. chrysocephala) and a greater incidence of Pieris rapae larvae, but reduced the extent of grazing by pigeons and slugs. Decreasing the side chain length of aliphatic glucosinolates and reducing the extent of hydroxylation of butenyl glucosinolates increased the extent of adult flea beetle feeding. The implications of modifying the glucosinolate content of the leaves of oilseed rape and the role of these secondary metabolites in plant/herbivore interactions are discussed.     

Effectiveness of the defence mechanism of the turnip sawfly, Athalia rosae (Hymenoptera: Tenthredinidae), against predation by lizards

The turnip sawfly, Athalia rosae Linnaeus, is a pest on cruciferous crops. Larvae sequester secondary plant compounds, namely glucosinolates, in their haemolymph. When attacked, their integument is easily disrupted and a droplet of haemolymph is exuded ('easy bleeding'). This has been shown to be an effective, chemical-based, defence against invertebrate predators. The efficiency of this proposed defence was tested against a vertebrate predator, using groups of the iguanid lizard Anolis carolinensis Voigt as a model predator. Caterpillars of Pieris rapae Linnaeus and Pieris brassicae Linnaeus served as control prey species that do not sequester glucosinolates. Lizards attacked far fewer sawfly larvae than pierid caterpillars. Several of the sawfly larvae were rejected after an initial attack, demonstrating unpalatability to the lizards, while the Pieris larvae were not rejected. However, P. rapae larvae topically treated with extracts of haemolymph of A. rosae had no deterrent effect on the lizards and no avoidance learning occurred over a period of two weeks. Adult sawflies do not easy bleed but have glucosinolates carried over from the larval stage. Lizards attacked them at a higher rate than larvae and they were never rejected. The results suggest that for the defensive effectiveness of the pest sawfly species against vertebrates the chemical cue is not necessarily sufficient. Movement and colour may be important additional factors triggering the behaviour of vertebrate predators.     

Cross habituation to feeding deterrents and acceptance of a marginal host plant byPieris rapae larvae

Sensitivity of the cabbage butterfly,Pieris rapae L. to feeding deterrents was compared for larvae reared on different food sources under laboratory conditions. Since cabbage-reared larvae normally reject nasturtium,Tropaeolum majus L., the effects of previous exposure to allelochemicals on larval acceptance or rejection of this plant were also examined. When compared with cabbage-reared larvae, nasturtium-reared larvae were less sensitive to feeding deterrents including cymarin, erysimoside and 2-O-β-d-glucosyl cucurbitacin E. Nasturtium-reared larvae were insensitive to chlorogenic acid, which was deterrent to cabbage-reared larvae. Feeding by larvae reared on a wheat germ diet was not deterred by these compounds. The results indicate that dietary experience can extensively affect larval sensitivity to feeding deterrents and that cross habituation of larvae to deterrents occurs in response to certain chemical constituents of nasturtium and wheat germ diet. Digitoxin, however, proved to be an exception. Larvae reared on either nasturtium or wheat germ diet were as sensitive to digitoxin as those reared on cabbage. Previous results have shown that rejectionof nasturtium by cabbage-reared larvae is due to the presence of strong feeding deterrents in this plant. However, more than 50% of 2nd instar larvae reared from neonate on cabbage leaves treated with strophanthidin, cymarin, erysimoside, digitoxigenin and digitoxin accepted nasturtium as a food source. 2-O-β-d-glucosyl cucurbitacin E, 2-O-β-d-glucosyl cucurbitacin I and rutin were also active in causing larvae to feed on nasturtium. Thus dietary exposure to unrelated plant chemicals can profoundly affect insect acceptance of a plant that contains feeding deterrents.     

Absence of red-light enhancement of phototropism in pea seedlings at limiting irradiances of blue light

The effect of different light qualities (blue, green, white, red and far-red) on ethylene production in leaf discs and flower petal discs of Begonia × hiemalis cv. Schwabenland Red was studied. All the light qualities, except far-red, reduced the ACC-conversion to ethylene in leaf discs by about 70% at a photosynthetic photon flux density (PPFD) of 20 mol m^–2s^–1.Blue and green light were less inhibitory than white and red light at lower PPFD. In all treatments far-red light at 0.5 mol m^–2s^–1 of photon flux density (PFD) stimulated the ACC-conversion to ethylene in leaf discs by about 60–90% compared to the dark-incubated control. White and red light strongly inhibited the -naphthalene-acetic acid (NAA) stimulated ethylene synthesis in leaf discs. The results may suggest that the ethylene production is controlled by phytochrome in the leaves but not in the petals. Lack of coaction of any light quality with silver ions on ethylene production in leaf and petal discs was also observed.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - EFE ethylene forming enzyme - NAA -naphthalene-acetic acid - PFD photon flux density - PPFD photosynthetic photon flux density - RH relative air humidity - SAM S-adenosylmethionine - STS silver thiosulphate     

Food mixing by generalist grasshoppers: Plant secondary compounds structure the pattern of feeding

Plant secondary compounds can prevent feeding by phytophagous insects or, if ingested, can be poisonous to them. Less attention has been paid to the additional effects they have on feeding behavior when they are only weakly deterrent or not deterrent at all. Experiments were carried out on the generalist grass-hopperSchistocerca americana. Individuals were presented either with two cakes of high-quality artificial food with a single deterrent compound added or with two cakes, each with a different added deterrent compound. The deterrents consisted of single plant secondary compounds that were either marginally or strongly deterrent. There were profound differences in feeding behavior between those individuals given identical and those given different cake types, including longer feeding bouts on single cakes when a choice of different cake types was available. The behavioral effects demonstrate that the presence of secondary compounds in one food can influence the patterns of feeding on other available foods and suggest that such chemicals could impact foraging activities in a complex manner.     

Natural Plant Chemicals Acting as Oviposition Deterrents on Cabbage Butterflies (Pieris btassicae (L.), P. rapae (L.) and P. napi (L.))

In several growers' reports Solatium lycopersicum, Sambucus nigra. Thymus vulgaris, Salvia officinalis, Artemisia absinthium, A. abrotanum , and Allium cepa are said to decrease the oviposition of Pieris brassicae, P. rapae and P. napi. In the present study the butterflies were fed with honey automats and reared throughout the year in artificial light in an insec-tarium. In a dual-choice chamber with a slow throughflow of air two equally sized cabbage leaves were placed on opposite sides. Significantly fewer eggs were layed on the cabbage leaf on which extracts of the mentioned plants had been applied. Ten butterflies were used in each experiment. The chemoreceptors and the chemicals involved are not identified but the inhibitory substances are surely secondary plant substances. Acceptance or rejection of secondary plant metabolics determines the complicated food relationships between plants and insects. The use of secondary plant substances for ecological control of insect pests is proposed.     

Ability of a generalist insect,Schistocerca gregaria,to overcome thioglucoside defense in desert plants: tolerance or adaptation?

Desert locusts, Schistocerca gregaria (Forskål) (Orthoptera, Acrididae), are generalist herbivores that, in the Sahara desert, may at times feed only on Schouwia purpurea (Forskål) (Brassicaceae), that is 10 times richer in thioglucosides than currently observed in other crucifers (>100 moles/g d.w.). Thioglucosides, when ingested, release products that are usually toxic to generalist insects.We studied the short-term (8 days) and long-term (21–26 days) consequences of a Schouwia-only diet on the digestion of these insects. The response was compared to the effects of a diet of Brassica oleracea, a crucifer well consumed in laboratory rearing conditions (7 moles/g glucosinolates d.w.).We found that the production of a myrosinase was induced in the midgut as early as 8 days following exposure to glucosinolates. No negative short-term effects were observed on the growth of the insect, but the activity of -glucosidases decreased in the midgut. The long-term exposure to the Schouwia diet affected activities of -glucosidases and -galactosidases, growth and assimilation efficiency. The limited adaptation of the desert locust to plant glucosides is compensated by an ability to tolerate high concentrations of allelochemicals for a short period.     

The effect of varying alkaloid concentrations on the feeding behavior of gypsy moth larvae, Lymantria dispar (L.) (Lepidoptera: Lymantriidae)

Nine alkaloids (acridine, aristolochic acid, atropine, berberine, caffeine, nicotine, scopolamine, sparteine, and strychnine) were evaluated as feeding deterrents for gypsy moth larvae (Lymantria dispar (L.); Lepidoptera: Lymantriidae). Our aim was to determine and compare the taste threshold concentrations, as well as the ED₅₀ values, of the nine alkaloids to determine their potency as feeding deterrents. The alkaloids were applied to disks cut from red oak leaves (Quercus rubra) (L.), a plant species highly favored by larvae of this polyphagous insect species. We used two-choice feeding bioassays to test a broad range of biologically relevant alkaloid concentrations spanning five logarithmic steps. We observed increasing feeding deterrent responses for all the alkaloids tested and found that the alkaloids tested exhibited different deterrency threshold concentrations ranging from 0.1 to 10 mM. In conclusion, it appears that this generalist insect species bears a relatively high sensitivity to these alkaloids, which confirms behavioral observations that it avoids foliage containing alkaloids. Berberine and aristolochic acid were found to have the lowest ED₅₀ values and were the most potent antifeedants. Handling Editor: Joseph Dickens.     

Differential selection of host plants by twoPieris species: the role of oviposition stimulants and deterrents

Oviposition responses ofPieris rapae L. andP. napi oleracea Harris to nine crucifers, one Capparidaceae and one Tropaeolaceae were directly compared under controlled conditions. Chemical fractions from these plants were also tested on both insects for the presence of oviposition stimulants or deterrents. The results showed that plant chemistry is a key factor in differential selection of potential hosts by thesePieris species. Some plant species were equally acceptable to bothPieris species. However,P. rapae preferred cabbage over most test plants whereasP. napi oleracea strongly preferred plant species that were avoided byP. rapae. The observed preferences were explained in most cases by the presence of stimulants and deterrents in extracts of the plants. The twoPieris species have apparently evolved differential sensitivities to the chemical stimuli that trigger or deter oviposition. The balance of positively and negatively interpreted sensory signals evoked by plant chemicals obviously plays an important role in acceptance or rejection of a plant by both species. The role of specific glucosinolates and differing structure-activity relationships is suggested.     

The role of an epipharyngeal sensillum in the perception of feeding deterrents by Leptinotarsa decemlineata larvae

An epipharyngeal taste sensillum in Leptinotarsa decemlineata larvae was studied. Electron microscopy showed that the sensillum is innervated by five neurons. Electrophysiological experiments showed that one of these cells responds to water, a second to sucrose and a third to two feeding deterrents that were also effective in a behavioural test. Receptor cells sensitive to feeding deterrents were not previously reported for L.␣decemlineata larvae or adults. The response of the sucrose-sensitive cell was strongly inhibited by one of the two feeding deterrents and only slightly by the other feeding deterrent. The relationship between the behavioural and electrophysiological results is discussed in order to elucidate the neural code of feeding deterrents in L. decemlineata larvae. We conclude that probably both the response of the deterrent cell and peripheral interactions exerted by feeding deterrents on the sucrose-sensitive cell determine the potency of feeding deterrents. The present results provide a physiological basis for the hypothesis that the presence or absence of feeding deterrents in potential food plants is a decisive cue in food plant selection by L. decemlineata larvae. Accepted: 25 March 1998     

Specificity of induced resistance in wild radish: causes and consequences for two specialist and two generalist caterpillars

Inducible plant resistance against herbivores is becoming a paradigm of plant–herbivore ecology. Fundamental to understanding induced resistance and its evolutionary ecology is specificity of "induction" and "effects". Specificity in the induction of resistance refers to whether plant damage by various herbivores causes the same response in plants. Specificity in the effects of induced resistance refers to whether induction has the same consequences (i.e., reduced preference or performance) for various herbivores. I examined both specificity of induction and effect employing four lepidopteran herbivores and wild radish plants, a system for which fitness benefits and costs of induction have been documented for the plant. Variation in the specificity of induction and effects of induced plant resistance was found; however, this variation was not associated with diet specialization in the herbivores (i.e., specialists vs generalists). Induction caused by Plutella (specialist) and Spodoptera (generalist) resulted in general resistance to all of the herbivores, induction caused by Pieris (specialist) induced resistance only to Spodoptera (generalist) and Pieris , and plant damage by Trichoplusia (generalist) failed to induce resistance and reduce the performance of any of the herbivores. To the contrary, plants damaged by Trichoplusia supported enhanced growth of subsequently feeding Trichoplusia compared to uninduced controls. These results add a novel level of complexity to interactions between plants and leaf chewing caterpillars. Within the same guild of feeders, some herbivores cause strong induced resistance, no induced resistance, or induced susceptibility. Similarly, caterpillar species were variable in the level to which induced resistance affected their performance. Such interactions limit the possibility of pairwise coevolution between plants and herbivores, and suggest that coevolution can only be diffuse.