Endophytic insect communities of two prairie perennials (Asteraceae: Silphium spp.)

Little is known of the biology of most insects that are endemic to prairie ecosystems of North America, with the exception of large and conspicuous species. In particular, species that are sequestered within plant tissues are commonly overlooked. In this paper, we assess the biodiversity of endophytic insects that inhabit stems of Silphium laciniatum L. and S. terebinthinaceum Jacquin (Asteraceae), endemic plants of tallgrass prairies. Endophytic herbivores, gall wasps Antistrophus rufus Gillette and A. minor Gillette (Hymenoptera: Cynipidae) and stem-boring larvae of the beetle Mordellistena aethiops Smith (Coleoptera: Mordellidae) were attacked by 10 species of natural enemies. We report new host plant associations for herbivores, and new host insect associations for parasitoids. The two plant species differed significantly in their densities of gall wasps and the vertical dispersion of galls within stems. Interactions within and between trophic levels attest to the biodiversity of endophytic insect communities, and the specialized nature of these insects suggests they are highly vulnerable to habitat conservation practices that involve destruction of dead vegetation.     

Populations of Predaceous Natural Enemies Developing on Insect-Resistant and Susceptible Tomato in North Carolina

Naturally occurring populations of immature and adultGeocoris punctipes,adultColeomegilla maculataand immature coccinellids were monitored on field-grown tomato lines susceptible and resistant toManduca sextaandHelicoverpa zea. Helicoverpa zeaandHeliothis virescenseggs and small larvae that serve as prey for these predators also were monitored. MoreH. zeaandH. virescenseggs and small larvae were found on resistant than on susceptible plant lines. However, similar populations of largeH. zeaandH. virescenslarvae were found on resistant and susceptible plants. The number of adultGeocoris punctipes,adultColeomegilla maculataand immature coccinellids on resistant plants was always as high or higher than the number on susceptible plants. The data demonstrate no incompatibility of host-plant resistance with biological control provided by these predaceous insects, but indicate that the number ofG. punctipesand coccinellids required to provide effective biological control may develop too late in the season to be of practical value. Large populations of stilt bugs (Jalysus wickhami,Hemiptera: Berytidae) and spiders were observed to occur earlier in the growing season than eitherG. punctipesor coccinellids and may be a significant source of mortality forH. zeaeggs and small larvae.     

The effect of ozone fumigation and different Brassica rapa lines on the feeding behaviour of Pieris brassicae larvae

Elevated levels of tropospheric ozone and their effects on plants have been studied for a great number of years. Ozone is a gaseous pollutant and acts as a phytotoxin. Even though ozone is known to change the physiology of plants, little attention has been given to the indirect effects of ozone on plant-insect interactions. This paper addresses this question by investigating the interactive effects of ozone and plant genotype on insects. Lines of rapid-cycling Brassica rapa (L.) selected for their contrasting sensitivity to ozone and Pieris brassicae (L.) (Lepidoptera: Pieridae) were used as a model system. The effect of differences in ozone sensitivity and ozone fumigation on the plant's carbon and nitrogen pools, the feeding preference, and behaviour of P. brassicae larvae were investigated. The results show that the plant's susceptibility to ozone interacts in a complex way with ozone induced alterations in the suitability of the plant for the insect. Only the larval performance on the sensitive line was affected by ozone exposure. Biochemical changes in the resistant B. rapa line made the plant a better food source for the insects, since the digestibility of this plant was significantly higher than that of the sensitive line, and the larvae pupated more quickly and were heavier.     

Impact of foliar herbivory on the development of a root-feeding insect and its parasitoid

The majority of studies exploring interactions between above- and below-ground biota have been focused on the effects of root-associated organisms on foliar herbivorous insects. This study examined the effects of foliar herbivory by Pieris brassicae L. (Lepidoptera: Pieridae) on the performance of the root herbivore Delia radicum L. (Diptera: Anthomyiidae) and its parasitoid Trybliographa rapae (Westwood) (Hymenoptera: Figitidae), mediated through a shared host plant Brassica nigra L. (Brassicaceae). In the presence of foliar herbivory, the survival of D. radicum and T. rapae decreased significantly by more than 50%. In addition, newly emerged adults of both root herbivores and parasitoids were significantly smaller on plants that had been exposed to foliar herbivory than on control plants. To determine what factor(s) may have accounted for the observed results, we examined the effects of foliar herbivory on root quantity and quality. No significant differences in root biomass were found between plants with and without shoot herbivore damage. Moreover, concentrations of nitrogen in root tissues were also unaffected by shoot damage by P. brassicae larvae. However, higher levels of indole glucosinolates were measured in roots of plants exposed to foliar herbivory, suggesting that the development of the root herbivore and its parasitoid may be, at least partly, negatively affected by increased levels of these allelochemicals in root tissues. Our results show that foliar herbivores can affect the development not only of root-feeding insects but also their natural enemies. We argue that such indirect interactions between above- and below-ground biota may play an important role in the structuring and functioning of communities.     

Electrohydrodynamic effects on two species of insects with economic importance to stored food products

An electrohydrodynamic (EHD) system which generated air ions within a strong electric field was used to study responses of stored-product insectsTribolium confusum (du Val) andPlodia interpunctella (Hübner). Larval mortality of both species generally increased with increased exposure time to ions of either polarity. The larvae and pupae ofT. confusum suffered a higher mortality rate than the adults. The insects initially exhibited distinct avoiding motions away from regions of high towards low fluxes of air ions of both polarity. Insects moved vigorously, tumbled, flipped, curled up, and aggregated when the EHD system was turned on. The control insects not exposed to air ions survived and showed a total absence of such behaviour. For bipolar exposures, the insects occupied the neutral zone where the effects were minimal due to cancellation of the fields. Prolonged exposures of more than 20 min produced a quiescent state. EHD-enhanced mass transfer of the liquid component from physical objects established in fluid mechanics was invoked as a possible cause for insect mortality and avoiding behaviour to ions. Body fluid losses increased linearly with time of exposure (R ²0.97) for all biological stages of insect growth. The larvae and pupae ofT. confusum lost 12 and 15% of their body fluids, respectively, after 80 min of exposure to negative air ions. Fluid losses of such a magnitude are likely to have contributed to insect fatality.     

Multiple insect resistance in transgenic tomato plants over-expressing two families of plant proteinase inhibitors

Protease inhibitors have been proposed as potential defense molecules for increased insect resistance in crop plants. Compensatory over-production of insensitive proteases in the insect, however, has limited suitability of these proteins in plant protection, with very high levels of inhibitor required for increased plant resistance. In this study we have examined whether combined used of two inhibitors is effective to prevent this compensatory response. We show that leaf-specific over-expression of the potato PI-II and carboxypeptidase inhibitors (PCI) results in increased resistance to Heliothis obsoleta and Liriomyza trifolii larvae in homozygote tomato lines expressing high levels (#62;1 the total soluble proteins) of the transgenes. Leaf damage in hemizygous lines for these transformants was, however, more severe than in the controls, thus evidencing a compensation response of the larvae to the lower PI concentrations in these plants. Development of comparable adaptive responses in both insects suggests that insect adaptation does not entail specific recognition of the transgene, but rather represents a general adaptive mechanism triggered in response to the nutritional stress imposed by sub-lethal concentrations of the inhibitors. Combined expression of defense genes with different mechanisms of action rather than combinations of inhibitors may then offer a better strategy in pest management as it should be more effective in overcoming this general adaptive response in the insect.     

Transgenic broccoli expressing aBacillus thuringiensis insecticidal crystal protein: Implications for pest resistance management strategies

We usedAgrobacterium tumefaciens to transform flowering stalk explants of five genotypes of broccoli with a construct containing the neomycin phosphotransferase gene and aBacillus thuringiensis (Bt) gene [CryIA(c) type] optimized for plant expression. Overall transformation efficiency was 6.4%; 181 kanamycin-resistant plants were recovered. Of the 162 kanamycin-resistant plants tested, 112 (69%) caused 100% morality of 1st-instar larvae of aBt-susceptible diamondback moth strain. Southern blots of some resistant transformants confirmed presence of theBt gene. Selected plants that gave 100% mortality of susceptible larvae allowed survival of a strain of diamondback moth that had evolved resistance toBt in the field. F₁ hybrids between resistant and susceptible insects did not survive. Analysis of progeny from 26 resistant transgenic lines showed 16 that gave segregation ratios consistent with a single T-DNA integration. Southern analysis was used to verify those plants possessing a single T-DNA integration. Because these transgenic plants kill susceptible larvae and F₁ larvae, but serve as a suitable host for resistant ones, they provide an excellent model for tests ofBt resistance management strategies.     

Host Plant Phenotype and the Impact and Development ofCarmenta mimosa,a Biological Control Agent ofMimosa pigrain Australia

The influence of host plant phenotype on the impact caused toMimosa pigraL. (Mimosaceae) plants by the feeding activity of larvae ofCarmenta mimosaEichlin and Passoa (Lepidoptera, Sesiidae) and the relationship between plant phenotype (through plant quality) and larval development were investigated under controlled conditions. Plants grown under conditions of reduced light availability were most prone to the detrimental effects of the feeding activities ofC. mimosalarvae. The relative growth rate (RGR) of most plant phenotypes could be reduced when infested withC. mimosalarvae compared to uninfested plants. Small plants infested withC. mimosaand grown under reduced light availability exhibited significant reductions in RGR sooner than large plants grown in full sun. The physical quality of stems was correlated with the severity of impact, i.e., whether stem breakage occurred. Little larval-induced mortality of plants was observed, suggesting that vascular tissues remained sufficiently intact to allow translocation. Systemic stem death arose through infection by secondary pathogens. Differences in biomass of infested versus uninfested plants of all phenotypes were associated with the loss of stems and to a lesser extent leaves. The phenotype of plants had a significant effect on the development ofC. mimosalarvae. Larvae in stems of plants with good access to reserves of soil moisture reached more advanced stages of development sooner than did those in plants which were often water stressed. This response was mediated through the combined influence of availability and nutritional quality of food. The implications of these findings to the impact of this agent in the field and the factors which may significantly influence its population dynamics are discussed.     

Role of plant phenology in mediating interactions between two biological control agents for spotted knapweed

The role of spotted knapweed phenology on the attack rate of two seed-head insects Urophora affinis and Larinus minutus was assessed in a series of field studies at four study sites in south-eastern British Columbia, Canada. Slow or later developing knapweed plants had more seed heads that contained only single or multiple U. affinis whereas early or faster developing plants had more seed heads containing L. minutus alone or in combination with U. affinis. L. minutus did not distinguish between seed heads with or without U. affinis larvae when laying eggs. However, seed heads with multiple U. affinis present, produced fewer L. minutus adults than expected. The probability of single or multiple U. affinis galls being present increased with seed-head diameter but was not affected by seed-head height. Attack by L. minutus increased with seed-head diameters >5 mm and was lower at plant heights above 50 cm. These results demonstrate two mechanisms that enable U. affinis to successfully coexist with L. minutus: differences between the species in their response to the developmental phenology of knapweed heads, and increased survivorship of U. affinis in heads with multiple U. affinis galls through niche interference competition. These mechanisms provide a possible explanation for the persistence of U. affinis populations on spotted knapweed, in spite of high levels of within seed-head mortality that have been observed with increasing L. minutus populations.     

Season long intraplant epizootics of entomopathogens,Beauveria bassiana andNosema pyrausta,in a corn agroecosystem

Intraplant epizootics of entomopathogens,Beauveria bassiana (Balsamo) Vuillemin, andNosema pyrausta (Paillot) were studied in a corn,Zea mays L., agroecosystem. Egg masses of the European corn borer,Ostrinia nubilalis (Hübner), infected withN. pyrausta were placed on midwhorl-stage corn plants. Conidia ofB. bassiana were applied in an aqueous suspension. Frass from the initial insects remaining within the plants was contaminated with sufficientN. pyrausta spores to infest 80% of the filial generation in each year of a 2-yr study. Viability ofN. pyrausta within the frass was monitored throughout the winter. Potential for impact of this inoculum on the filial generation is discussed. Conidia from cadavers of the European corn borer that were killed by the initial inoculum ofB. bassiana and/or conidia from the initial inoculum ofB. bassiana significantly reduced tunneling by the filial generation of the European corn borer. However, neither the concentration ofB. bassiana nor the age of the larvae exposed toB. bassiana, had any significant (P<.05) effect on tunneling by the 2nd-generation larvae. This paper reports the results of research only. Mention of a pesticide in this paper does not constitute a recommendation for use by the U.S. Department of Agriculture or cooperators, nor does it imply registration under FIFRA as amended.     

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.     

Comparative headspace analysis of cabbage plants damaged by two species ofPieris caterpillars: consequences for in-flight host location byCotesia parasitoids

Headspace composition, collected from intact cabbage plants and cabbage plants infested with eitherPieris brassicae L. orP. rapae L. (Lepidoptera: Pieridae) first instar larvae, was determined by GC-MS. Twenty-one volatiles were identified in the headspace of intact plants. Twenty-two volatiles were identified in the headspace of plants infested byP. brassicae larvae, 2 of which, Z-3-hexenyl butyrate and Z-3-hexenyl isovalerate, were not detected in the headspace of either intact orP. rapae damaged plants. In the headspace of the latter, 21 compounds were identified, all of which which were also produced by intact plants. No significant quantitative differences were found between headspace composition of the plants damaged by one or the other caterpillar species. Major differences between intact and caterpillar-damaged plants in contribution to the headspace profile were revealed for hexyl acetate, Z-3-hexenyl acetate, myrcene, sabinene and 1,8-cineole. The larval endoparasitoidCotesia glomerata L. was attracted by the volatiles emanating fromB. oleracea damaged byP. brassicae first instar larvae.C. rubecula L., a specialized larval endoparasitoids ofP. rapae, was attracted by the volatiles released from theB. oleracea-P. rapae plant-host complex. This shows that cabbage plants kept under the conditions of headspace collection produce attractive volatiles for both parasitoids.     

Monophagy in a polyphagous grasshopper,Schistocerca shoshone

The feeding behavior of different populations of the grasshopper,Schistocerca shoshone, was investigated in the southwestern United States. Insects from three riparian populations, with a broad spectrum of plants available to them, tended to eat plants roughly in relation to their availability except that broad-leaved herbaceous plants were avoided. Insects from a desert population in a plantation ofSimmondsia fed exclusively on that plant, as did those from another population in the Tucson mountains, despite the availability of a range of other plants. Insects from a third desert population, near Portal, fed mainly onProsopis, the dominant woody plant. In detailed behavioral experiments in the laboratory, insects from Tucson mountains readily acceptedSimmondsia, and less readily acceptedProsopis. Three other common woody plants from the habitat were generally rejected without feeding. Insects from Portal acceptedProsopis andSimmondsia with approximately equal readiness. Breeding experiments suggested that the differences between the plantation insects and those from Portal was genetic and not induced by experience. The insects from both populations were potentially polyphagous and ate a wide range of plants in the laboratory if given no alternative.     

Predator Presence Moves <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> Larvae to Distraction

Displacement of herbivorous insects by the presence of predators on whole plants has rarely been studied. By semi-continuous observations of an externally feeding insect herbivore and a predator, we show how the mere presence of the predator, Geocoris lubra Kirkaldy (Hemiptera: Geocoridae), on a plant can have a strong influence on the movement and behaviors of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) larvae. The presence of predators, as opposed to mortality by predators, influenced the proportion of larvae feeding, resting and implementing avoidance activities. In addition, the proportion of time individual larvae allocated to feeding, resting and dropping off plants was affected when predators were present with and without contact between the two. Predators do more than just reduce numbers of herbivores; they influence feeding, displacement and subsequently the distribution of plant damage.     

Performance of leaf beetle larvae on sympatric host and non-host plants

Studies asking the ability of insects to utilize novel host plants often use novel hosts that are allopatric with the insect population under investigation. However, since the outcomes of species interactions are often site-specific, such studies cannot tell us whether a plant would actually be used by a given insect population if the plant grew sympatrically with it. We therefore performed a quantitative genetics experiment to analyse the performance of larvae of the leaf beetle Oreina elongata Suffrian (Coleoptera: Chrysomelidae, Chrysomelinae) on two host and three non-host plants, collected from a site where insects and plants co-occur in the Western Alps. When raised on the non-host Petasites albus (L.), larvae were able to survive equally well as on the two hosts, Adenostyles alliariae (Gouan) and Cirsium spinosissimum (L.), whereas they did not survive on the two other non-hosts, Peucedanum ostruthium (L.) and Rumex alpinus L. On P. albus, growth rate was slightly lower and development time slightly longer than on the two hosts. We found a genotype by environment interaction only for growth rate but not for development time and survival. However, the shape of the reaction norms of growth rates suggests that it is unlikely that selection could favour the inclusion of P. albus into the host range of the study population.     

Predators mediate host plant resistance to a phytophagous pest in cabbage with glossy leaf wax

This study examined the role of generalist predators in producing higher mortality ofPlutella xylostella L. (Plutellidae) larvae on glossy vs. normal-wax cabbage,Brassica oleracea var.capitata L. To test this, survival and feeding ofP. xylostella were measured on individually caged glossy and normal-wax plants with and without each of three generalist predators,Chrysoperla carnea (Stephens) (Chrysopidae),Orius insidiosus (Say) (Anthocoridae), andHippodamia convergens Guerin-Meneville (Coccinellidae). In the greenhouse, predators always significantly reduced survival ofP. xylostella larvae on glossy plants, but never on normal-wax plants. In the field, predators significantly reducedP. xylostella survival on glossy plants, but onlyC. carnea was effective on normal-wax plants. In similar experiments with excised leaves,O. insidiosus andC. carnea were more effective predators on the glossy leaves, whileH. convergens was equally effective on both kinds of leaves. Patterns for feeding were similar, but significance levels differed from those forP. xylostella survival. The greater effectiveness of predators on glossy plants is apparently due to the reported improved mobility of these animals on glossy leaf surfaces. The data also suggest that reduced mining byP. xylostella exposes the larvae to more predation on glossy plants and contributes some to the resistance. Regardless of the mechanism, resistance toP. xylostella on glossyB. oleracea appears to depend on the action of generalist predators for its full expression. This dependence on predation must be considered in the development and deployment of glossy insect-resistantB. oleracea.     

The foraging behaviors of larvae of the ladybird beetle, <Emphasis Type="Italic">Coccinella septempunctata</Emphasis> L., (Coleoptera: Coccinellidae) towards ant-tended and non-ant-tended aphids

The foraging behaviors of larvae of the ladybird, Coccinella septempunctata L., towards both the ant-tended aphid, Aphis craccivora Koch, and the non-ant-tended aphid, Acyrthosiphon pisum Harris, were investigated in the field and in laboratory experiments. Although there were no differences in the development and growth of the ladybird larvae that preyed on either Ac. pisum or Ap. craccivora, the foraging efficiency of the ladybird larvae that preyed on Ap. craccivora was higher than that of the ladybird larvae that preyed on Ac. pisum in the absence of ants. This result was explained by the fact that the number of Ac. pisum that escaped by dropping off the plant was conspicuously larger than the number of Ap. craccivora that escaped in this fashion and derived from the non-ant-attendance associated with Ac. pisum. In the laboratory experiments, fewer ladybird larvae climbed onto a plant with Ap. craccivora in the presence of ants than onto a plant with Ac. pisum in the absence of ants. The ladybird larvae did not switch from foraging for Ap. craccivora to foraging for Ac. pisum, even after suffering attacks by ants on a plant with Ap. craccivora, and it would appear that ladybird larvae are unable to remember where they have previously been attacked by ants. These results could explain why the ladybird larvae in the field more frequently visited Vicia angustifolia plants with Ap. craccivora than those with Ac. pisum and made more visits when ants were absent than when they were present.     

Predation by three hemipterans:Tropiconabis nigrolineatus,Oechalia schellenbergii andCermatulus nasalis,onHeliothis punctiger larvae in two types of searching arenas

Three species of hemipteran predators preyed differently upon 1^st instarHeliothis punctiger Wallengren larvae.Cermatulus nasalis consumed more larvae thanOechalia schellenbergii which consumed more larvae thanTropiconabis nigrolineatus. All the species consumed significantly less 1^st instar larvae on plants than what they consumed in Petri-dishes. Fifth instar predators showed significant differences in terms of prey consumption due to sex independent of searching conditions. Only 4^th and 5^th instars ofT. nigrolineatus attacked and captured 2^nd instars ofH. punctiger larvae. The other 2 species however readily attacked and consumed 2^nd instarH. punctiger larvae. Their prey consumption was similar in Petri-dishes and on plants. Only 5^th instars ofT. nigrolineatus could subdue and capture 3^rd instarH. punctiger larvae. Second instar pentatomids captured just one 3^rd instar larva but older instars killed and ate more. Fourth instarH. punctiger larvae were immune to attacks by allT. nigrolineatus and younger pentatomids due to their defense ploys but 5^th instar pentatomids could subdue and capture them. None of the predators captured 5^th instarH. punctiger larvae except few 5^th instar females ofC. naslis andO. schellenbergii.      

Maintenance of narrow diet breadth in the monarch butterfly caterpillar: response to various plant species and chemicals

In order to better understand the maintenance of a fairly narrow diet breadth in monarch butterfly larvae, Danaus plexippus L. (Lepidoptera: Nymphalidae: Danainae), we measured feeding preference and survival on host and non-host plant species, and sensitivity to host and non-host plant chemicals. For the plant species tested, a hierarchy of feeding preferences was observed; only plants from the Asclepiadaceae were more or equally preferred to Asclepias curassavica, the common control. The feeding preferences among plant species within the Asclepiadaceae are similar to published mean cardenolide concentrations. However, since cardenolide data were not collected from individual plants tested, definitive conclusions regarding cardenolide concentrations and plant acceptability cannot be made. Although several non-Asclepiadaceae were eaten in small quantities, all were less preferred to A. curassavica. Additionally, these non-Asclepiadaceae do not support continued feeding, development, and survival of first and fifth-instar larvae. Preference for a host versus a non-host (A. curassavica versus Vinca rosea) increased for A. curassavica reared larvae as compared to diet-reared larvae suggesting plasticity in larval food preferences. Furthermore, host species were significantly preferred over non-host plant species in bioassays using a host plant or sucrose as a common control. Larval responses to pure chemicals were examined in order to determine if host and non-host chemicals stimulate or deter feeding in monarch larvae. We found that larvae were stimulated to feed by some ubiquitous plant chemicals, such as sucrose, inositol, and rutin. In contrast, several non-host plant chemicals deterred feeding: caffeine, apocynin, gossypol, tomatine, atropine, quercitrin, and sinigrin. Additionally the cardenolides digitoxin and ouabain, which are not in milkweed plants, were neutral in their influence on feeding. Another non-milkweed cardenolide, cymarin, significantly deterred feeding. Extracts of A. curassavica leaves were tested in bioassays to determine which components of the leaf stimulate feeding. Both an ethanol extract of whole leaves and a hexane leaf-surface extract are phagostimulatory, suggesting the involvement of both polar and non-polar plant compounds. These data suggest that the host range of D. plexippus larvae is maintained by both feeding stimulatory and deterrent chemicals in host and non-host plants.     

In vivo andin vitro investigations on rotenoids fromIndigofera tinctoria and their bioefficacy against the larvae ofAnopheles stephensi and adults ofCallosobruchus chinensis

Various plant parts ofIndigofera tinctoria L. were collected separately at different growth stages and analysed for their rotenoid content. The total rotenoid content decreased with age; among the plant parts, maximum content was in leaves and minimum in stem. The identity of different rotenoids was confirmed by melting point, mixed melting point, UV and infrared spectral studies, and gas-liquid chromatography. Six rotenoids (deguelin, dehydrodeguelin, rotenol, rotenone, tephrosin and sumatrol) were isolated, identified and quantified invivo. The static cultures ofIndigofera tinctoria were established from seeds on RT medium, and maintained for a period of six months by frequent subculturings. Only four rotenoids were present in callus cultures; sumatrol and tephrosin were absent. The maximum content was found in eight week old tissue after fresh subculturings and minimum at 2 weeks. The toxicological studies ofin vivo andin vitro extract against the pulse beetle(Callosobruchus chinensis) and mosquito(Anopheles stephensi) larvae, showed that rotenoids were more effective against mosquito larvae thanCallosobruchus chinensis. Extracts from callus was more effective against both the test animals than that from plant parts.