Behavioural and sensory responses to some neem compounds by Pieris brassicae larvae

Abstract. The efficacy of a commercial product (Margosan-O) to inhibit feeding in Pieris brassicae L. (Lepidoptera: Pieridae) larvae was compared to that of three pure triterpenoids of botanical origin: azadirachtin and salannin (occurring in Melia azedurach ), and toosendanin (occurring in M. toosendan ). In dual-choice behaviour tests the order of antifeedant efficacy was: Margosan-O > toosendanin > salannin > azadirachtin. The ranking order of their capacity to stimulate the deterrent receptor located in the medial sensillum styloconicum appears to be: Margosan-O = toosendanin > salannin > azadirachtin. In addition to stimulating the medial deterrent cell, toosendanin inhibits the sugar receptor and the glucosinolate receptor, both located in the lateral sensillum styloconicum. The amino acid receptor in this sensillum, however, is not affected. In contrast, neither azadirachtin nor salannin influences the sensitivity of any of the receptor cells in this sensillum.
The adaptation rate of the deterrent receptor is low in comparison to that of the two sugar receptors.
A significant correlation is found between the antifeedant indices of the four neem compounds tested at three different concentrations, and the responses of the deterrent receptor to these compounds at similar concentrations. It is concluded that when this insect species feeds on its natural foodplant (i.e. cabbage), the deterrent effect of neem compounds is mediated solely via the medial deterrent receptor, whereas inhibitory effects on the sugar and glucosinolate receptors do not play a significant role.     

Behavioural and sensory responses to drimane antifeedants in Pieris brassicae larvae

Fifteen drimane compounds were tested for their feeding inhibiting activity in larvae of Pieris brassicae L. (Lepidoptera: Pieridae) when applied to leaf material of the host plant Brassica oleracea L. The antifeedant efficacy of the drimanes was related to their molecular structure in order to identify important functional groups. Of the drimanes tested, those with a lactone group on the B-ring were the most effective feeding inhibitors. Additionally, the sensory responses to 13 of the drimanes were measured. Neural activity was evoked in the deterrent cell in the medial sensillum styloconicum. Also, inhibition of sensory responses to feeding stimulants was found. Results of behavioural and electrophysiological tests were correlated in an attempt to elucidate the sensory code underlying feeding inhibition by drimanes in Pieris brassicae. It was concluded that the response of the deterrent cell in the medial sensillum styloconicum contributes significantly to inhibition of feeding behaviour in larvae of Pieris brassicae.     

Comparative study on the responses of maxillary sensilla styloconica of cotton bollwormHelicoverpa armigera and Oriental tobacco budwormH. assulta larvae to phytochemicals

Using the electro-physiological technique, the sensory mechanisms of maxillary sensilla styloconica to stimulants and deterrents were explored on two closely related species, the generalistHelicoverpa armigera and the specialistH. assulta. The results showed that: (i) in both species, cells sensitive to sucrose and azadirachtin were mainly in the lateral sensillum styloconicum, and those to inositol were in the medial sensillum styloconicum; (ii) sensitivity of medial sensillum styloconicum inH. assulta to inositol was higher than that inH. armigera; (iii) among 6 tested deterrents, only azadirachtin evoked high impulse discharge from the lateral sensillum styloconicum in both insects; (iv) the deterrents could disturb stimulants evoking impulse discharge from maxillary sensilla styloconica of both species in different degrees: To sucrose evoking impulses on lateral sensillum styloconicum, forH. armigera capsaicin had a strong inhibition and gossypol had a weak inhibition, while forH. assulta tannic acid, gossypol, and tomatine all had strong inhibition except nicotine and capsaicin; to inositol evoking impulses on medial sensilum styloconicum, forH. armigera inhibition of tomatine was strong but that of gossypol was weak; and forH. assulta inhibition of gossypol was strong but that of nicotine was weak.     

家蚕味觉电生理反应的个体差异

为探讨家蚕Bombyx mori人工饲料饲养发育不齐的生理原因,从同一蚕品种中挑选出对人工饲料摄食性不同的个体,用电生理学方法测定了下颚瘤状体味觉感器对4种代表性物质（蔗糖、肌醇、大豆粉提取物和柠檬酸）的电生理反应。结果表明,栓锥感器Ss-Ⅰ对蔗糖等促食物质的反应以及栓锥感器Ss-Ⅱ对大豆粉提取物等阻食物质的反应,均存在明显的个体差异。在临界浓度下,低摄食性个体的放电脉冲频率显著高于高摄食性个体。说明低摄食性蚕的味觉反应比高摄食性蚕敏感。     

Comparative study on the responses of maxillary sensilla styloconica of cotton bollwormHelicoverpa armigera and Oriental tobacco budwormH. assulta larvae to phytochemicals

Using the electro-physiological technique, the sensory mechanisms of maxillary sensilla styloconica to stimulants and deterrents were explored on two closely related species, the generalist Helicoverpa armigera and the specialist H. assulta. The results showed that: (i) in both species, cells sensitive to sucrose and azadirachtin were mainly in the lateral sensillum styloconicum, and those to inositol were in the medial sensillum styloconicum; (ii) sensitivity of medial sensillum styloconicum in H. assulta to inositol was higher than that in H. armigera; (iii) among 6 tested deterrents, only azadirachtin evoked high impulse discharge from the lateral sensillum styloconicum in both insects; (iv) the deterrents could disturb stimulants evoking impulse discharge from maxillary sensilla styloconica of both species in different degrees: To sucrose evoking impulses on lateral sensillum styloconicum, for H. armigera capsaicin had a strong inhibition and gossypol had a weak inhibition, while for H. assulta tann     

An electrophysiological analysis of the effect of phagostimulant mixtures on the responses of a deterrent-sensitive cell of gypsy moth larvae, Lymantria dispar (L.)

Gypsy moth larvae, Lymantria dispar (L.), are polyphagous feeders. The medial styloconic sensillum of this species bears a taste receptor cell that responds to alkaloids and another that responds to the sugar alcohol, inositol. The lateral styloconic sensillum bears a taste receptor cell that is sensitive to the sugar, sucrose. We tested the effect of two phagostimulants, namely sucrose and inositol, on the response of the deterrent-sensitive cell and found that both phagostimulants suppressed its response, equally, while their combination was significantly more effective. We also tested the effect of two alkaloids (i.e., strychnine and caffeine), which deter feeding in this species, on the response of the inositol- and sucrose-sensitive cells. Although both of these deterrents had no effect in suppressing the response of the sucrose-sensitive cell, they both had an effect in suppressing the inositol-sensitive cell. We also found that sucrose suppressed the response of the inositol-sensitive cell, whereas inositol had no significant effect on the response of the sucrose-sensitive cell. In this paper, we examined the effect of mixtures of these compounds to determine the nature of their interaction. In the context of host–plant interactions and, for example, host recognition, whereby host plant acceptability depends on the total sensory impression acquired from responses to multiple plant components rather than the presence or absence of single stimulant or deterrent compounds, this study could have a direct bearing in the development of natural compounds (i.e., alkaloids) for pest control and crop protection. It will also contribute to our understanding of the neural basis of the feeding behavior of this insect.     

草地贪夜蛾幼虫对四种刺激物质的味觉感受和取食选择

【目的】为了筛选有效的草地贪夜蛾Spodoptera frugiperda幼虫取食激食素和抑制剂并探究其味觉感受机理,为生态防治草地贪夜蛾提供理论和实践上的依据。【方法】利用单感受器记录法测定草地贪夜蛾5龄第2天幼虫下颚外颚叶上中栓锥感器和侧栓锥感器对不同浓度的蔗糖、黑芥子苷、单宁酸和盐酸奎宁4种刺激物质的电生理反应,并采用二项叶碟法测定草地贪夜蛾幼虫对这些刺激物质的取食选择行为。【结果】草地贪夜蛾幼虫中栓锥感器和侧栓锥感器内均存在对蔗糖、黑芥子苷和单宁酸敏感的味觉受体神经元,但是神经元的活性随着刺激物的种类及浓度而变化。其中,两类感器内神经元对蔗糖和黑芥子苷的反应均呈现典型的浓度梯度反应。中栓锥感器内存在对盐酸奎宁敏感的味觉受体神经元,但是呈现逆浓度梯度的反应模式,侧栓锥感器内不存在对盐酸奎宁敏感的神经元。蔗糖显著诱导幼虫的取食行为,而盐酸奎宁、黑芥子苷和单宁酸均抑制幼虫的取食行为,且都呈现浓度梯度的抑制活性。【结论】草地贪夜蛾幼虫中栓锥感器和侧栓锥感器内均存在对取食激食素和抑制剂敏感的味觉受体神经元,但是两类感器不论在反应谱上还是敏感性上均存在差异。蔗糖可以作为取食激食素,盐酸奎宁...     

The Anti-Botulism Triterpenoid Toosendanin Elicits Calcium Increase and Exocytosis in Rat Sensory Neurons

Toosendanin, a triterpenoid from Melia toosendan Sieb et Zucc, has been found before to be an effective anti-botulism agent, with a bi-phasic effect at both motor nerve endings and central synapse: an initial facilitation followed by prolonged depression. Initial facilitation may be due to activation of voltage-dependent calcium channels plus inhibition of potassium channels, but the depression is not fully understood. Toosendanin has no effect on intracellular calcium or secretion in the non-excitable pancreatic acinar cells, ruling out general toosendanin inhibition of exocytosis. In this study, toosendanin effects on sensory neurons isolated from rat nodose ganglia were investigated. It was found that toosendanin stimulated increases in cytosolic calcium and neuronal exocytosis dose dependently. Experiments with membrane potential indicator bis-(1,3-dibutylbarbituric acid)trimethine oxonol found that toosendanin hyperpolarized capsaicin-insensitive but depolarized capsaicin-sensitive neurons; high potassium-induced calcium increase was much smaller in hyperpolarizing neurons than in depolarizing neurons, whereas no difference was found for potassium-induced depolarization in these two types of neurons. In neurons showing spontaneous calcium oscillations, toosendanin increased the oscillatory amplitude but not frequency. Toosendanin-induced calcium increase was decreased in calcium-free buffer, by nifedipine, and by transient receptor potential vanilloid 1 (TRPV1) antagonist capsazepine. Simultaneous measurements of cytosolic and endoplasmic reticulum (ER) calcium showed an increase in cytosolic but a decrease in ER calcium, indicating that toosendanin triggered ER calcium release. These data together indicate that toosendanin modulates sensory neurons, but had opposite effects on membrane potential depending on the presence or absence of capsaicin receptor/TRPV 1 channel.     

Morphological and electrophysiological characteristics of the epipharyngeal sensilla of the silkworm, Bombyx mori

One pair of gustatory sensilla was found on the epipharynx ofBombyx mori larvae, and some morphological and electrophysiological characteristics of the epipharyngeal sensilla were investigated. They are sensilla coeloconica composed of a small papilla with a pore at the tip and a swelling of cuticle encircling the papilla. Three bipolar neurons innervate each sensillum. One neuron is an inositol receptor which responds to inositol only. Another cell responds with action potentials of relatively large amplitude to some feeding deterrent substances, such as strychnine nitrate. The thresholds of these cells for inositol and strychnine nitrate are approximately 10⁻⁴ M and 10⁻⁷ M, respectively. At least two kinds of spikes can be observed when these sensilla are stimulated with some salts and acids. Dose-response relationships and time courses of responses to inositol and strychnine nitrate were also examined in this study.     

Behavioral and chemosensory responses to a host recognition cue by larvae of <Emphasis Type="Italic">Pieris rapae</Emphasis>

Larvae of the cabbage white Pieris rapae are specialists on plants belonging to the family Brassicaceae (Cruciferae). Adult females have been shown to use the glucosinolate gluconasturtiin (phenylethylglucosinolate) as a recognition cue for cruciferous plants, so they can identify an appropriate host for oviposition (Huang and Renwick in J Chem Ecol 20:1025–1037, 1994). Here, we report our results from a study of the role of this glucosinolate in feeding preferences of P. rapae larvae. The larvae were allowed to choose between leaf disks from the non-host cowpea Vigna sinensis (Fabaceae) that were treated with pure gluconasturtiin in solvent, or solvent alone. Our results showed that gluconasturtiin is a feeding stimulant for P. rapae larvae. A series of chemosensory ablations revealed that this response is mediated by one set of taste sensilla, the sensilla styloconica. Electrophysiological tip recordings revealed two neurons in the lateral sensillum styloconicum that are sensitive to gluconasturtiin. These neurons show significantly higher firing frequencies with 4 mM gluconasturtiin added to the recording pipette than for recording solution alone. We propose that the sensitivity to gluconasturtiin shown by these two taste neurons is an important contributor to the animals behavioral preference for this compound.     

Some properties of gustation in the larva of Pieris brassicae

The effects of some chemical substances on feeding response were studied in the larvae of Pieris brassicae L. Some synergistic effects between feeding stimulants were found. The larvae appeared to be highly sensitive to deterrents. In the medial sensillum styloconicum of the maxillae a chemoreceptor is present which is stimulated by a wide range of specific feeding inhibitors among which many belong to the alkaloids. Some physiological properties of the deterrent sensitive receptor are given and its significance in food selection behaviour is discussed.

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Résumé Les effets de quelques substances chimiques sur la prise de nourriture ont été étudiés chez la chenille de Pieris brassicae. Des effets synergiques se manifestent entre certains acides aminés ou certaines vitamines et le saccharose, lorsque ces produits sont à des concentrations appropriées. Par ailleurs les chenilles se montrent très sensibles à la présence de substances inhibant la prise de nourriture. Un neurone chimiorécepteur appartenant au sensillum styloconicum médian des maxilles se révèle sensible à une grande variété de ces substances inhibitrices, dont la plupart appartiennent aux alcaloides stéroides. Ce récepteur est sensible aux composés voisins de l'ecdysone présents dans certaines plantes, et son rôle physiologique est discuté, en rapport avec le comportement alimentaire et le choix de la nourriture.

     

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     

Triterpenoid saponins stimulate the sugar taste receptor cell through a G protein-mediated mechanism in the blowfly,Phormia regina

The blowfly has taste chemosensilla on the labellum. The sensory receptor cells in the chemosensillum are highly specialized for the tastes of sugar, salt and water, respectively. Previously we introduced chromosaponin I (CSI) and glycyrrhizin (GL), as sweet substances for the blowfly, Phormia regina. Application of these triterpenoid saponins induced feeding responses as well as impulses of the sugar taste receptor cell in the LL-type sensillum at a much lower concentration than that of sucrose. In the present paper, we show the involvement of G protein-mediated cascade in the CSI- and GL-responses as well as in sugar responses. CSI activates the sugar signal transduction cascade after penetrating through the membrane. On the other hand, GL exerts dual effects to stimulate the sugar signal transduction possibly by activating it inside the cell and also by interacting with the pyranose sugar receptor site. A non hydrolyzable G protein inhibitor guanosine 5′-O-(2-thiodiphosphate), GDPβS, markedly decreased the responses of the sugar receptor cell to the two triterpenoid saponins as well as the response to sucrose and fructose. These results suggest that CSI and GL are direct activators of G protein.     

Gustatory perception of phytoecdysteroids in Plodia interpunctella larvae

Phytoecdysteroids are steroidal compounds produced by various plants that disrupt growth and development of insects eating them. They exhibit an insecticidal activity on a number of insect pests such as Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae). In this study, we further evaluated whether phytoecdysteroids deter larvae of this species from feeding, by using four phytoecdysteroid molecules, commonly occurring in plants: 20‐hydroxyecdysone (20E), ponasterone A (PonA), polypodine B (PolB), and makisterone A (MakA). Fourth instar P. interpunctella avoided contact with food pellets treated with these phytoecdysteroids in a dose‐dependent way (2–30 mm ). In order to test whether this avoidance was mediated by taste sensitivity, we recorded the responses of taste neurons located in the lateral and medial sensilla styloconica of the galea. At least one neuron responded to each of these compounds in both sensilla. The neuron located in the medial sensillum had a detection threshold of 10^?6 m for PonA, 10^?4 m for 20E and PolB, and 10^?3 m for MakA. The lateral sensillum neuron responded with less intensity and its detection threshold was 10 times higher than that of the medial sensillum neuron. These results indicate that phytoecdysteroids are detected as deterrent stimuli by P. interpunctella larvae and that small structural differences significantly affect their biological activity.     

Responses of the African Armyworm and three species of borers to carbohydrates and phenolic substances: an electro- and behavioural physiological study

Electrophysiological recordings were made from the maxillary sensilla styloconica of fifth instars of the African Armyworm, Spodoptera exempta (Wlk.) (Lep.: Noctuidae), and the borers (Lep.: Pyralidae) Eldana saccharina Wlk. and Maruca testulalis (Geyer), and fifth and sixth instars of the borer Chilo partellus (Swinhoe). Stimuli were different concentrations of 13 carbohydrates and 11 phenolic substances. It appeared that a sucrose sensitive cell is present in the lateral styloconic sensilla of Spodoptera, Maruca and Chilo, and in both the lateral and medial sensilla of Eldana. The sucrose cell of fifth-instar Chilo is more sensitive than that of sixth instars. The additional responses of the medial cells to the phagostimulant sucrose are supposed to have contributed to the widespread infestation of sugarcane by Eldana. The medial sensilla of the Armyworm contain a cell type responding to both meso-inositol and D-ribose. Chilo appeared to be the only one of the three borer species able to perceive phenolic substances. It has a cell responsive to chlorogenic acid in its medial sensilla, which is a bit more sensitive in fifth- than in sixth-instar larvae. The presence of this cell may account for the positive correlation between total phenols content and resistance of sorghum cultivars to this borer. Spodoptera can perceive four phenolic acids. Its lateral sensilla contain a cell responsive to chlorogenic acid and its medial sensilla a cell responsive to 4-hydroxybenzoic acid, gallic acid as well as vanillic acid. Feeding experiments showed that the latter substances inhibit feeding in the Armyworm.So far, in the borers adequate stimuli could be determined for only tow or three of the four taste cells present in each sensillum styloconicum. This indicates that we are still unaware of chemicals that may be important to these caterpillars. On the other hand, for the sensilla styloconica of Spodoptera, electrophysiology suggests the presence of five or six taste cell types, whereas only four taste cells are known from morphological studies.     

Deterrence coding by a larval Manduca chemosensory neurone mediating rejection of a non-host plant, Canna generalis L.

Abstract. The physiological basis of phagodeterrence was studied electrophy-siologically and behaviourally in the phytophagous caterpillars Manduca sexta and Manduca quinquemaculata. The model unacceptable non-host plant was the canna lily, Canna generalis. A strongly deterrent extract was obtained from fresh leaves of canna by extraction with hot ethanol or ethyl acetate in a blender. Behavioural rejection of these extracts was similar to that of fresh leaves, although less intense. In contrast, blender extracts using other solvents, as well as leaf surface rinses, were phagostimulant or neutral. Chromatographic fractionation of the deterrent ethanolic extract showed the active principles to be moderately polar and separable into two fractions. Previous ablation experiments had shown that the medial maxillary styloconica and epipharyngeal sensilla are the two most important chemosensory organs in mediating behavioural rejection of canna leaves; if only one of these organs is spared, the animal completely rejects canna. We investigated the neural responses of the medial styloconica and their contribution to the sensory coding responsible for this phagodeterrence. The active fractions of the deterrent ethanolic extract elicited a vigorous response from one chemosensory neurone in the medial styloconica. This neurone is distinguishable from others in the medial styloconica by its unique temporal response parameters and the characteristic shape changes of its action potentials. The response frequency of this neurone correlates with the degree of phagodeterrence in a dose-dependent manner. Threshold deterrence occurs at a concentration of extract (1%) that elicits firing in this neurone at a rate of c. 50 spikes/s peak instantaneous frequency and 30 total spikes in the first Is. We conclude that this is a ’deterrent neurone’ in the sense that vigorous response from this neurone is a sufficient sensory code for behavioural rejection of canna. Thus input from a single sensory neurone is capable of blocking feeding, since only one (unilateral) medial styloconicum is needed to mediate this rejection.     

Taste cell responses in the polyphagous arctiid, Grammia geneura: towards a general pattern for caterpillars

The caterpillars of Grammia geneura are polyphagous as individuals. Electrophysiological responses of its medial and lateral galeal styloconic sensilla to 21 amino acids, 6 carbohydrates, 10 chemically diverse plant secondary compounds and two inorganic salts were examined. In the medial sensillum, a single cell responded to 8 amino acids, 3 carbohydrates, and the iridoid, catalpol, which is present in a favored hostplant. In the lateral sensillum, one cell responded to amino acids and another to fructose. Two cells in each sensillum responded to secondary compounds and it is suggested that the same cells are stimulated by inorganic salts. There was no evidence of a separate salt-sensitive cell. Phenylalanine stimulated a deterrent cell in the medial sensillum and was behaviorally deterrent. Some essential amino acids did not stimulate any cells and it is suggested that a small number of amino acids (sometimes non-essential) may serve as indicators of nutrient quality. Sugars probably serve as the primary phagostimulants because they are in relatively high concentrations in plants. It is proposed that taste receptor cells should be categorized primarily by their behavioral effects as phagostimulatory or deterrent, rather than their specific ranges of responsiveness. This would emphasize the basic similarities across taxa.     

Chemoreception and feeding behaviour in a caterpillar: towards a model of brain functioning in insects

The quantitative relationship between chemoreceptor activity during food intake and total amount of food ingested per unit of time has been studied in last instar larvae of Pieris brassicae. Data on sensory responses and feeding intensity, derived from a study by Blom (1978), show a strong correlation between receptor activity during 1 sec periods of stimulation and food intake during 24 h. It is concluded that nerve impulses from chemoreceptors, which signal the presence of different phagostimulants, are summated algebraically in the central nervous system. Impulses from feeding deterrent receptors counteract the effects of phagostimulants. One impulse from a deterrent receptor neutralizes 2.5 impulses from phagostimulant receptors. Based on the relationships observed a simple model of a feeding centre in the central nervous system has been constructed (Fig. 8).