Antifeedant and toxic activity of Trichilia americana extract against the larvae of Spodoptera litura

The biological activity of a crude methanolic extract of Trichilia americana (Sesse and Mocino) Pennington (Meliaceae) was assessed using the Asian armyworm, Spodoptera litura (Fabr.) (Lepidoptera: Noctuidae). The extract exhibited strong antifeedant activity in a choice leaf disc bioassay with 0.18 g cm^–2 extract deterring feeding by 50%. In nutritional assays, the crude extract reduced growth, consumption and the utilisation of ingested and digested food in a dose-dependent manner when fed to larvae, suggesting both antifeedant and toxic activities. When relative growth rates were plotted against relative consumption rates, the growth efficiency of the S. litura fed on diet containing T. americana crude extract was significantly less than that of control larvae. This result further indicates that the extract acts as both an antifeedant and chronic toxin. Toxicity is only seen following ingestion and was not observed following topical application or injection into the hemocoel. Larvae reared initially on extract-containing diet then transferred to control diet showed nutritional indices comparable to those of larvae fed continuously on control diet. This suggests that the extract is not permanently damaging the insect's digestive tract. The mode-of-action of the extract as a chronic toxin remains unknown.     

The effect of neem allelochemicals on nutritional physiology of larval Spodoptera litura

Neem allelochemicals azadirachtin, salannin, nimbinene and nimbin were administered to different larval instars of the tobacco armyworm, Spodoptera litura orally in artificial diet, topically or via injection. Nutritional analyses revealed strong antifeedant and growth regulatory effects of azadirachtin which were independent of each other. While salannin and nimbinene induced concentration dependent feeding deterrence only; nimbin was inactive to the 1000 ppm level against this insect species. One of the causes of the reduced growth rate of azadirachtin treated insects was due to an increase in the costs associated with growth. This was relative to a drastic reduction in the activity of gut trypsin. Salannin and nimbinene, however, did not interfere with the trypsin activity of the gut. These results and those from nutritional studies suggest that salannin and nimbinene have no toxicity mediated effects on S. litura larvae and antifeedant activity is a result of the effects on deterrent and other chemoreceptors. The fact that azadirachtin directly or indirectly inhibits the secretion of trypsin by the enzyme-secreting cells of the gut is discussed.     

Bioefficacy and mode-of-action of aglaroxin A from Aglaia elaeagnoidea (syn. A. roxburghiana) against Helicoverpa armigera and Spodoptera litura

The bioefficacy of aglaroxin A from Aglaia elaeagnoidea (syn. A. roxburghiana) was assessed using the gram pod borer, Helicoverpa armigera (Hübner), and Asian armyworm, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae). The compound exhibited strong growth inhibition in a diet bioassay, with 0.67 p.p.m: and 0.78 p.p.m. of the compound reducing growth by 50% in H. armigera and S. litura neonate larvae, respectively, whereas a growth inhibition of 95% was achieved at 2.36 p.p.m: and 2.41 p.p.m., respectively; this was comparable to azadirachtin treatments used as a control. Aglaroxin A was toxic to various stadia. Nutritional analysis revealed the antifeedant properties of the compound; however, nutritional indices indicated that the reduction in growth of the larvae was not entirely due to starvation, but partly due to the toxic effects of the ingested compound. This was further confirmed in topical treatments. When relative growth rate was plotted against relative consumption rate, the growth efficiency of larvae fed on a diet containing aglaroxin A was significantly less than that of control larvae. These results further indicate that aglaroxin A acts as both antifeedant and chronic toxin. Morphologically deformed or partially pupated insects were obtained after 5th instar larvae were treated with aglaroxin A. Such developmental inhibition during ecdysis was not due to depletion of the moulting hormone, as treated larvae, when provided with exogenous 20‐hydroxyecdysone, did not show any recovery from the effect. However, it is obvious from the present findings that aglaroxin A activity does not absolutely follow the pattern of azadirachtin or the more related compound rocaglamide known in lepidopterans.     

Repellent,antifeedant and toxic effects of Ageratum conyzoides (Linnaeus) (Asteraceae) extract against Helicovepra armigera (Hübner) (Lepidoptera: Noctuidae)

Repellent, antifeedant and toxic effect of crude hexane extract of Ageratum conyzoides were investigated against Helicoverpa armigera. In orientation bioassay, the extract exhibited dose-dependent repellency against neonates. Extract significantly increased the mortality and decreased growth of different larval stages when administrated orally in artificial diet. EC₅₀ value was at 0.11% for larval growth inhibition. Toxicity of the extract was manifested by high mortality of first instar larvae after 7 days of feeding on diet containing 0.05–0.4% of extract with LC₅₀ of 0.17%. Under choice bioassay, extract showed strong antifeedant activity against fifth instar larvae with DI₅₀ of 0.21%. In nutritional bioassay, extract significantly reduced RCR, RGR, ECI and ECD of fifth instar larvae with increased AD. When RGR were plotted against RCR, the growth efficiency of larvae fed on treated diet was significantly lower than the control fed larvae suggesting the antifeedant and toxic effect of extract.     

Selective Feeding of <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> (Hübner) and <Emphasis Type="Italic">Spodoptera litura</Emphasis> (Fabricius) on Meridic Diet with <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> Toxins

Laboratory experiments were conducted to evaluate the behavior of Helicoverpa armigera (Hübner) and Spodoptera litura (Fabricius) larvae on meridic diet with different concentrations of Bt spray formulation Delfin or isolated Cry1Ac protein or the foliage and bolls from transgenic cotton, Bollgard hybrid RCH-317 Bt. Both insect species selectively fed on nontreated diet compared with the diet treated with Delfin. While H. armigera exhibited concentration response with Cry1Ac, this protein did not affect S. litura larvae. In general Helicoverpa selected diet with low concentrations (EC₂₀ and EC₅₀ levels) of Cry1Ac compared with higher concentrations of Cry1Ac. In order to develop appropriate management strategies, a thorough understanding of the behavioral mechanisms leading to the responses of insects to the proteins in transgenic varieties is required. Thus, based on results of the insects fed individually on the leaf discs or bolls from transgenic cotton plants alone or under choice situation with meridic diet revealed that H. armigera larvae preferred meridic diet to transgenic leaves or bolls expressing Cry1Ac protein. H. armigera larvae preferred meridic diet to plant material; more than 70% larvae were seen on the meridic diet, and average larval weight gain was in the range of 121.7–130.5 mg. However, in case of S. litura the larvae showed no significant discrimination between meridic diet and the leaf discs. In fact more than 60% larvae preferred leaf discs for feeding, though Cry1Ac expression in leaf discs was in the range of 0.9–2.18 μg/g. Thus differences in behavioral response could potentially impact the level of efficacy of crop cultivars that have been genetically engineered to produce these proteins.     

Antifeedant and larvicidal activities of Rhein isolated from the flowers of Cassia fistula L.

Antifeedant and larvicidal activities of rhein (1,8-dihydroxyanthraquinone-3-carboxylic acid) isolated from the ethyl acetate extract of Cassia fistula flower were studied against lepidopteron pests Spodoptera litura and Helicoverpa armigera. Significant antifeedant activity was observed against H. armigera (76.13%) at 1000 ppm concentration. Rhein exhibited larvicidal activity against H. armigera (67.5), S. litura (36.25%) and the LC₅₀ values was 606.50 ppm for H. armigera and 1192.55 ppm for S.litura. The survived larvae produced malformed adults.     

Insect feeding deterrents from the medicinal plant Tylophora asthmatica

Alkaloid extractables of Tylophora asthmatica (Asclepiadaceae) have been tested for the antifeedant activity against Spodoptera litura by incorporating them in semisynthetic and cellulose diets. The alkaloid extractables at 0.01% concentration inhibited feeding of the larvae. Three compounds, tylophorine, tylophorinine and pergularinine, were isolated; and their EC₅₀ values were found to be 2.9, 8.6 and 12.00 ppm, respectively.     

Avidin expressed in transgenic tobacco leaves confers resistance to two noctuid pests,Helicoverpa armigera and Spodoptera litura

Fertile transgenic tobacco plants with leaves expressing avidin in the vacuole have been produced and shown to halt growth and cause mortality in larvae of two noctuid lepidopterans, Helicoverpa armigera and Spodoptera litura. Late first instar H. armigera larvae and neonate (<12-h-old) S. litura larvae placed on leaves excised from T₀ tobacco expressing avidin at 3.1–4.6M (moles/kg of fresh leaf tissue) had very poor growth over their first 8 days on the leaves, significant numbers had died by days 11 or 12 and all were dead by day 22 (H. armigera) or day 25 (S. litura). Similar results were obtained when late first instar H. armigera larvae were placed on leaves from T₁ plants expressing avidin at six different average concentrations, ranging from 3.7 to 17.3M. Two larvae on the lowest expressing leaves survived to pupation, but there was total mortality among the other groups and no relationship between avidin concentration and the effects on the larvae. Synergistic effects between avidin-expressing tobacco plants and a purified Bt toxin, Cry1Ba, were demonstrated. Late instar H. armigera larvae fed with leaves from T₂ plants expressing avidin at average concentrations of either <5.3 or >12.9M, and painted with Cry1Ba protein at a rate equivalent to an expression level of 0.5% of total leaf protein, died significantly faster than larvae given either of the two treatments alone. Larvae fed with avidin-expressing leaves painted with the protease inhibitor, aprotinin, at a rate equivalent to 1% of total leaf protein had mortality similar to those given avidin-leaves alone. There was no evidence of antagonism between these two proteins.     

Cloning and expression inEscherichia coli of entomotoxic protein gene fromBacillus thuringiensis subspecieskurstaki

A plasmid borne larvicidal crystal protein gene from B.thuringiensis subspecieskurstaki was cloned inEscherichia coli using a specific 20-mer oligonucleotide probe. The gene expressed inE. coli at a high level. TransgenicE. coli cells produced large irregular bodies which looked bright under phase contrast microscopy. The phase bright bodies released by sonic disruption of cells could be pelleted by centrifugation. Toxicity trials on the larvae ofSpodoptera litura showed that the pellet was antifeedant and toxic to the larvae. The supernatant was only mildly antifeedant. Even short term feeding of larvae on the toxin delayed the onset of pupation.     

An improved leaf-disk antifeedant bioassay and its application for the screening of Hokkaido plants

In the course of our search for new insect antifeedant substances from plants, we have devised an improved antifeedant leaf-disk bioassay. Our system allows an accurate measurement of consumed disk surfaces, using a video camera interfaced with a computer. The scanned image of the leaf disks is stored, and the eaten areas are measured with the help of a video-image analysis software. This new method allows for precise quantification of insect antifeedant activity tested on leaf material. This method was applied to the screening of Hokkaido plants for antifeedant activity. Out of 206 samples, seven demonstrated significant activity against larvae of the tobacco cutworm,Spodoptera litura.     

Antifeedant effects of azadirachtin and structurally related compounds on lepidopterous larvae

The antifeedant activity of azadirachtin, azadirachtin-derivatives and related limonoids was assessed in choice and no-choice bioassays against four species of Lepidoptera: Spodoptera littoralis, Spodoptera frugiperda, Heliothis virescens and Heliothis armigera. The choice bioassay showed that the feeding behaviour of S. littoralis was affected by more of the compounds than that of either S. frugiperda or H. virescens. H. armigera was the least affected. Azadirachtin and dihydroazadirachtin were the most potent of the 40 compounds tested. The results showed that hydrogenation of the C-22,23 double bond did not decrease antifeedant activity and the nature of the substitutes at C-1, C-3 and C-11 were important. Molecules with bulky substitutes at either C-22 or C-23 were usually ineffective antifeedants as were compounds lacking an epoxide. Compounds recorded as active antifeedants in the choice bioassay were not always as active in the no-choice test. The value of the bioassays in assessing the mode of action of the compounds is discussed.

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Résumé L'activité phagodissuadante de l'azadirachtine, de ses dérivés et des limonoïdes voisins sur 4 espèces de lépidoptères: Spodoptera littoralis, S. frugiperda, Heliothis virescens et H. armigera a été évaluée par des expériences avec et sans choix. Les expériences de choix ont montré que le comportement alimentaire de S. littoralis était modifié par plus de substances que celui de S. frugiperda ou H. virescens. Celui de H. armigera était le moins modifié. Les 2 substances les plus puissantes parmi les 40 examinées, ont été l'azadirachtine et le dihydroazadirachtine. Ces résultats montrent que l'hydrogénation de la double liaison C-22,23 ne réduit pas l'activité phagodissuadante et que la nature des substitutions en C-1, C-3 et C-11 sont importantes. Les molécules avec des substitutions volumineuses en C-22 ou C-23 sont généralement des phagodissuadants aussi inefficaces que ceux ayant perdu un époxide. Les substances notées comme phagodissuadants actifs dans les expériences de choix ne sont pas toujours aussi actives en absence de choix. La valeur des tests dans l'évaluation du mode d'action des substances est discuté.

     

Synergistic interaction between sublethal doses of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> and <Emphasis Type="Italic">Campoletis chlorideae</Emphasis> in managing <Emphasis Type="Italic">Helicoverpa armigera</Emphasis>

Campoletis chlorideae Uchida (Hymenoptera: Ichneumonidae) is an important solitary larval endoparasitoid of the tomato fruit borer Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) in India. The interaction between Bacillus thuringiensis subspecies kurstaki (Btk) HD-1 and C. chlorideae was studied under laboratory condition to explore their compatibility in managing H. armigera. The results had indicated that the growth and development of H. armigera was affected in a dose-dependent manner upon feeding on sublethal doses of Btk HD-1-treated diets. There were no larval survivors in lethal doses of Btk HD-1 (LC₇₀ and LC₉₀). The growth and survival of the parasitoid were normal when the host larvae were fed with sublethal doses or subjected to short time exposure to lethal doses of Btk HD-1. However, the parasitoid offsprings developed slowly and pupal as well as adult period, adult weight and adult emergence rate were reduced significantly if the parasitoid was developing inside a severely Bt intoxicated host larvae. There were no evident differences in longevity of parasitoid adults that were fed on honey solution containing different concentrations of Btk HD-1 as compared to adults fed only on honey solution. This indicates no direct adverse effect of Btk HD-1 on C. chlorideae. Further, the gravid female parasitoid did not discriminate Btk HD-1 intoxicated and normal H. armigera larvae for oviposition. The result implies that spore crystal formulation of Btk HD-1 can be effectively used in a synergistic manner along with existing natural or prereleased population of C. chlorideae in organic farming or as components in biointensive IPM module for managing H. armigera.     

Secretory avocado idioblast oil cells: evidence of their defensive role against a non-adapted insect herbivore

We tested the hypothesis that avocado idioblast oil cells play a defensive role against herbivorous insects. Toxicities of the intact avocado idioblast oil cells and the extracted idioblast oil were compared for three insect herbivores. Spodoptera exigua (Hübner) larvae are generalists that do not feed on avocados. By contrast, Sabulodes aegrotata (Guenée) and Pseudoplusia includens (Walker) larvae are generalist herbivores that readily feed on avocados. All bioassays were performed at a naturally occurring concentration of idioblast oil cells (2% w/w). Choice experiments showed that S. exigua larvae avoided diet treated with avocado idioblast oil cells and consume more control than treated diet. In contrast, idioblast oil cells had no significant antifeedant effects on the adapted S. aegrotata and P. includens larvae. Subsequent experiments designed to assess resistance mechanisms separated pre-ingestive (behavioral) and post-ingestive (physiological) effects of the avocado idioblast oil cells, and the extracted idioblast oil, on the two adapted herbivores. Post-ingestive adaptation was the mechanism that allows feeding. Because the impact of the avocado idioblast oil cells was greatest on the performance of non-adapted S. exigua, additional experiments determined that larvae fed diet containing the oil cells had higher mortality and reduced larval growth compared to controls. Developmental times were significantly prolonged for the survivors. Thus, increased mortality, reduced developmental rates, and antifeedant activity in the non-adapted insect indicate that defense against non-adapted herbivores may be an important function of idioblast cells in avocados.     

Toxicity of the limonoid allelochemical cedrelone to noctuid larvae

The limonoid cedrelone, a constituent of Cedrela toona and C. odorata (Meliaceae) was evaluated as a larval growth inhibitor for the polyphagous noctuids Peridroma saucia and Mamestra configurata by examining its effects on development and feeding behaviour. Cedrelone significantly inhibited growth of different larval instars when administered orally (in artificial diet), topically or via injection. Nutritional analyses revealed that both growth inhibition and reduced consumption are a consequence of postingestive malaise rather than a peripherally-mediated antifeedant effect. Cedrelone administered via injection to sixth instar P. saucia larvae also inhibited growth, delayed development, and resulted in considerable larval mortality. However, all larvae which survived treatment pupated successfully. The present study suggests that the toxicity of cedrelone does not likely involve the endocrine system.     

Influence of transgenic cottons with <Emphasis Type="Italic">Bacillus thuringiensis cry1Ac</Emphasis> gene on the natural enemies of <Emphasis Type="Italic">Helicoverpa armigera</Emphasis>

Transgenic cotton has been released for cultivation in several parts of the world to increase crop productivity. However, concerns have been raised regarding the possible undesirable effects of genetically modified crops on non-target organisms in the eco-system. Therefore, we studied the effects of transgenic cottons with cry1Ac gene from Bacillus thuringiensis Berliner (Bt) on the natural enemies of cotton bollworm/legume pod borer, Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) under field and laboratory conditions. There was no apparent effect of transgenic cotton on the relative abundance of predatory spiders (Clubiona sp. and Neoscona sp.), coccinellid (Cheilomenes sexmaculatus Fab.), and the chrysopid (Chrysoperla carnea Stephens). However, the abundance of spiders, coccinellids, and chrysopids was quite low in insecticide protected plots towards end of the cropping season. There was a significant reduction in cocoon formation and adult emergence of the ichneumonid parasitoid, Campoletis chlorideae Uchnida reared on H. armigera larvae fed on the leaves of transgenic cottons before and after parasitization. However, no Bt toxins were detected in H. armigera larvae and the parasitoid cocoons with enzyme linked immunosorbent assay. Reduction in cocoon formation was because of early mortality of the H. armigera larvae due to Bt toxins in the leaves of transgenic cotton. There was a slight reduction in adult weight and fecundity, and prolongation of the larval period when the parasitoid was raised on H. armigera larvae fed on the leaves of transgenic cotton before and after parasitization. Survival and development of C. chlorideae was also poor when H. armigera larvae were fed on the leaves of cotton hybrid Mech 184. The adverse effects of transgenic cotton on survival and development of C. chlorideae were largely due to early mortality, and possibly poor nutritional quality of H. armigera larvae due to toxic effects of the transgene.     

Oviposition and reproductive performance of a generalist parasitoid (Trichogramma pretiosum) exposed to host species that differ in their physical characteristics

The response of generalist egg parasitoids to alternative natural hosts that are present simultaneously is not well known. We investigated the behavior of Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) in relation to two field hosts Helicoverpa armigera Hübner and Spodoptera litura Fabricius, in choice and no choice tests. We quantified the effects of natal host species and post-emergence adult age on the oviposition preference of the parasitoids. H. armigera eggs were consistently preferred over S. litura eggs, regardless of the natal host and adult age. When only S. litura eggs were available as hosts, they were parasitized at statistically similar rates to H. armigera eggs (average of 17 ± 2.7 vs. 13 ± 3.0, H. armigera to S. litura). The adult lifespan and lifetime fecundity of T. pretiosum were variable but were affected by natal host species and/or host species to which they were exposed. Mean lifespan and fecundity of parasitoids that had developed in H. armigera eggs and were exposed to H. armigera eggs for oviposition were 13.9 ± 1.8 days and 98.7 ± 11.0 adult offspring. By contrast, those that developed in S. litura eggs and were exposed to S. litura eggs for oviposition lived for 7 ± 0.9 days and produced 53.8 ± 8.0 adult offspring. The ovigeny index (OI) was significantly lower in the parasitoids exposed to H. armigera eggs than in those exposed to S. litura eggs, regardless of the natal host, indicating that H. armigera eggs sustain the adult parasitoids better than S. litura eggs. These results are used to predict parasitoid behavior in the field when both hosts are available.     

Novel strategies for the biocontrol of noctuid pests (Lepidoptera) based on improving ascovirus infectivity using Bacillus thuringiensis

Identifying novel biocontrol agents and developing new strategies are urgent goals in insect pest biocontrol. Ascoviruses are potential competent insect viruses that may be developed into bioinsecticides, but this aim is impeded by their poor oral infectivity. To improve the per os infectivity of ascovirus, Bacillus thuringiensis kurstaki (Btk) was employed as a helper to damage the midgut of lepidopteran larvae (Helicoverpa armigera, Mythimna separata, Spodoptera frugiperda, and S. litura) in formulations with Heliothis virescens ascovirus isolates (HvAV-3h and HvAV-3j). Btk and ascovirus mixtures (Btk/HvAV-3h and Btk/HvAV-3j) were fed to insect larvae (3rd instar). With the exception of S. frugiperda larvae, which exhibited low mortality after ingesting Btk, the larvae of the other tested species showed three types of response to feeding on the formulas: type I, the tested larvae (H. armigera) were killed by Btk infection so quickly that insufficient time and resources remained for ascoviral invasion; type II, both Btk and the ascovirus were depleted by their competition, such that neither was successfully released or colonized the tissue; type III, Btk was eliminated by the ascovirus, and the ascovirus achieved systemic infection in the tested larvae. The feeding of Btk/ascovirus formulas led to a great reduction in larval diet consumption and resulted in a significant decrease in the emergence rate of H. armigera, M. separata, and S. litura larvae, which suggested that the formulas exerted marked oral control effects on both the contemporary individuals and the next generation of these tested pest species.     

A Comparison of Growth and Development of Three Major Agricultural Insect Pests Infected with Heliothis virescens ascovirus 3h (HvAV-3h)

Ascoviruses are double-stranded DNA viruses that are pathogenic to lepidopteran hosts, particularly noctuid larvae. Infection of a larva is characterized by retarded growth, reduced feeding and yellowish body color. In this paper, we reported the growth and development of three major agricultural noctuid insect pests, Helicoverpa armigera (Hübner), Spodoptera exigua (Hübner) and Spodoptera litura (Fabricius), infected with Heliothis virescens ascovirus 3h (HvAV-3h). Using 10-fold serial dilutions (0 to 7) of HvAV-3h-containing hemolymph to infect S. litura larvae, we found no significant difference in larval mortalities from 0 to 10³-fold dilutions; however, significant differences were observed at 10⁴-fold dilution and above. Using a 10-fold dilution of HvAV-3h-containing hemolymph to infect H. armigera, S. exigua and S. litura larvae, we found that the growth and development were significantly affected. All infected larvae could not pupate; the survival times of treated H. armigera, S. litura and S. exigua larvae were significantly longer than untreated control larvae. Body weight showed significant difference between treated and untreated control group from day 1 after inoculation in H. armigera and S. exigua, but day 2 in S. litura. Additionally, food intake also showed significant difference between treated and untreated control group from day 2 after inoculation in H. armigera and S. litura, but day 3 in S. exigua.     

Food consumption and growth of larvae and juveniles of three cyprinid species at different food levels

Synopsis The relationships between food availability, consumption and growth were analyzed from the onset of feeding to an age of 90 days in three cyprinid species. Fish were held at 20 ± 0.5° C and given two (three) constant rations of approximately 30, (40) or 100% dry body weight (dbw) ind^-1 day^-1. Food consisted of living zooplankton, the size of which correlated with fish size. At high food densities consumption rates decreased rapidly with fish size in all three species. At reduced rations, fish consumed most of the food offered until they were larger than 10 mg dbw. In all species and at each feeding level daily rations consumed increased allometrically with body size. Respiration rate, expressed as routine metabolic rate differed insignificantly between the three species. At high ration levels, growth rates of small bleak, Alburnus alburnus, were distinctly lower than those of roach, Rutilus rutilus, and blue bream, Abramis ballerus. At low food supply all three species grew at similar rates. Assimilation efficiency at low food conditions was approximately twice that of the well-fed groups. If the caloric equivalents of oxygen consumption as measured in well-fed fish are applied to fish fed at low rations their energy budgets do not balance. This indicates the limitations of fish larvae in the partitioning of energy for growth or activity at such conditions.     

Expression of the cry1EC gene in castor (Ricinus communis L.) confers field resistance to tobacco caterpillar (Spodoptera litura Fabr) and castor semilooper (Achoea janata L.)

Castor (cv. DCS-9) has been transformed through Agrobacterium-mediated and particle gun bombardment methods using appropriate vectors containing the Bt chimeric gene cry1EC driven by enhanced 35S promoter. About 81 and 12 putative transformants were regenerated following selection on hygromycin and kanamycin, respectively. Southern analysis of DNA extracted from T₀ plants confirmed integration of the introduced gene in castor genome. The integration and inheritance of the introduced genes was demonstrated up to T₄ generation by PCR and Southern analysis. Southern analysis of two events having single and two copies showed the same pattern of integration in the subsequent generations. Insect feeding experiments conducted in the laboratory by releasing neonate larvae of castor semilooper and S. litura on leaf tissues excised from transgenic and control plants showed varying degrees of larval mortality and slow growth in larvae fed on transgenic leaf tissue. Field bioassays against Spodoptera litura and castor semilooper conducted for eight events in T₁–T₄ generations under net confinement were more informative and events conferring resistance to the two major defoliators were identified.