Plant‐mediated interactions among above‐ground and below‐ground life stages of a root‐feeding weevil

1. Interactions among herbivores mediated by plant responses to herbivore injury may have large impacts on herbivore population densities. Responses may persist for weeks after injury and may affect not only the initial (inducing) herbivore, but also herbivores that are spatially or temporally separated from the initial attacker. 2. In many plant–insect interactions, multiple life stages of the insect may be associated with the same plant, and these various stages may interact indirectly with one another via induced responses. The rice water weevil (RWW), Lissorhoptrus oryzophilus, a serious global pest of rice, is one such insect. A series of experiments were performed with root‐feeding larvae and leaf‐feeding adults of the RWW using three conventional rice varieties. 3. The first objective of this study was to test whether RWW adult feeding on rice leaves resulted in altered oviposition by subsequent adults. The hypothesis for the first objective was that RWW adult feeding would decrease plant suitability, resulting in reduced oviposition by subsequent adults. 4. The second objective was to test whether injury by RWW larvae to rice roots resulted in altered oviposition by subsequent adults. The hypothesis for the second objective was that below‐ground RWW larval feeding would decrease plant suitability of rice to above‐ground RWW adults, resulting in decreased oviposition. 5. Results provided inconsistent support for the first hypothesis, indicating that responses differed among combinations of variety and injury level. Conversely, consistent support for the second hypothesis was found, indicating that larval feeding on roots decreased suitability of rice plants for oviposition.     

Effects of defoliation on the resistance and tolerance of rice,Oryza sativa,to root injury by the rice water weevil,Lissorhoptrus oryzophilus

Understanding plant‐mediated interactions in agricultural systems may facilitate the development of novel and improved management practices, which is important, as management of these insects is currently heavily reliant on insecticides. The fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae, Prodeniini), is a sporadic pest of rice fields in the southern USA. In southwestern Louisiana, this defoliating insect typically attacks rice early in the growth season, before fields are flooded. Defoliation by fall armyworm larvae may trigger increased expression of plant defenses, which may result in increased resistance to subsequent herbivores. The rice water weevil (RWW), Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae, Stenopelmini), enters rice fields as an adult both before and after flooding, but oviposition and larval infestation occur only after fields are flooded. RWW may be affected by changes in plant resistance caused by fall armyworm defoliation before flooding. The objectives of this study were to investigate the plant‐mediated effects of natural and artificial defoliation on population densities of RWW larvae after flooding and on the ability of rice plants to compensate for root injury by RWW larvae. In the 2015 season, fall armyworm defoliation before flooding resulted in reduced RWW densities after flooding. However, in 2016 no significant effects of fall armyworm defoliation on densities of RWW larvae were detected. Similarly, mechanical defoliation of rice before flooding did not affect RWW densities after flooding. Although lowest yields were observed in plots subjected to both root injury and defoliation, there was little evidence of a greater than additive reduction in yields from simultaneous injury. These results suggest a lack of plant‐mediated interactions among these two pests in rice in the southern USA.     

Variation in damage to cotton affecting larval feeding preference of Spodoptera littoralis

Feeding experiments with larvae of Spodoptera littoralis were performed with leaves from cotton plants subjected to damage and from undamaged plants. In the experiments, four different time intervals (1, 3, 7, and 14 days) after damage induction and two different levels (high and low) of herbivore damage were tested. Seven days after damage induction larvae fed less on the young top leaves from damaged plants for both levels of damage. At the high damage level, the larvae fed less on leaves from the damaged plants after just three days, and this effect still remained 14 days after damage infliction. When mature leaves from the middle of the plant were compared, no difference between treatments was observed.Two plant sizes were tested, small plants with 4–5 true leaves and large plants with 8–10 true leaves. In small plants the induced changes affecting larval feeding were found mainly in the youngest leaf at the top of the plant, while in large plants the induced effects were found in both the youngest and the second youngest leaves.In plants subjected to artificial damage, larvae fed less on top leaves of the damaged plants when compared to leaves from undamaged plants. When leaves from plants that had been artificially damaged were directly compared with leaves from plants damaged by herbivores, larvae fed more on the youngest leaves from artificially damaged plants when the plants were large. In small plants no significant difference was found when comparing artificial and herbivore damage.     

Interactions of root and leaf herbivores on purple loosestrife (<Emphasis Type="Italic">Lythrum salicaria</Emphasis>)

Interspecific interactions of herbivores sharing a host plant may be important in structuring herbivore communities. We investigated host plant-mediated interactions of root (Hylobius transversovittatus) and leaf herbivores (Galerucella calmariensis), released to control purple loosestrife (Lythrum salicaria) in North America, in field and potted plant experiments. In the potted plant experiments, leaf herbivory by G. calmariensis reduced H. transversovittatus larval survival (but not larval development) but did not affect oviposition preference. Root herbivory by H. transversovittatus did not affect either G. calmariensis fitness or oviposition preference. In field cage experiments, we found no evidence of interspecific competition between root and leaf herbivores over a 4-year period. Our data suggest that large populations of leaf beetles can negatively affect root-feeding larvae when high intensity of leaf damage results in partial or complete death of belowground tissue. Such events may be rare occurrences (or affected by experimental venue) since field data differed from data obtained from potted plant experiments, particularly at high leaf beetle densities. Interspecific interactions between G. calmariensis and H. transversovittatus are possible and may negatively affect either species, but this is unlikely to occur unless heavy feeding damage results in partial or complete plant death.     

Adult beetle grazing induces willow trichome defence against subsequent larval feeding

Induced increases in trichome density to grazing by herbivores have been suggested to increase plant resistance to future herbivore attack. In this paper we present results which show that plants of Salix cinerea L. respond to adult leaf beetle (Phratora vulgatissima L.) grazing by developing new leaves with an increased trichome density. The same plants are usually attacked again later in the season when the next generation of larvae hatches on the plants. The effect of the induced response was studied by comparing larval growth and feeding on newly developed leaves of plants exposed to different defoliation treatments. Larvae on plants previously exposed to adult grazing consumed less total leaf area and showed more dispersed feeding than larvae on plants protected from previous grazing. Larvae on plants exposed to mechanical defoliation responded intermediately. These results corresponded to the increased trichome density of defoliated plants. However, we found this larval response only in whole plant tests--when reared on single, excised leaves in petri dishes, larvae in all treatments behaved similarly. This discrepancy between the on-plant experiment and that in petri dishes highlights how experimental design may alter the conclusion of a study. We suggest that the induced response to adult grazing may act as a defence against subsequent larval feeding.     

Larval parasitism of the autumnal moth reduces feeding intensity on the mountain birch

Plants respond to grazing by herbivorous insects by emitting a range of volatile organic compounds, which attract parasitoids to their insect hosts. However, a positive outcome for the host plant is a necessary precondition for making the attraction beneficial or even adaptive. Parasitoids benefit plants by killing herbivorous insects, thus reducing future herbivore pressure, but also by curtailing the feeding intensity of the still living, parasitised host. In this study, the effect of parasitism on food consumption of the 5th instar larvae of the autumnal moth (Epirrita autumnata) was examined under laboratory conditions. Daily food consumption, as well as the duration of the 5th instar, was measured for both parasitised and non-parasitised larvae. The results showed that parasitism by the solitary endoparasitoid Zele deceptor not only reduced leaf consumption significantly but also hastened the onset of pupation in autumnal moth larvae. On the basis of the results, an empirical model was derived to assess the affects on the scale of the whole tree. The model suggests that parasitoids might protect the tree from total defoliation at least at intermediate larval densities. Consequently, a potential for plant–parasitoid chemical signalling appears to exist, which seems to benefit the mountain birch (Betula pubescens ssp. czerepanovii) by reducing the overall intensity of herbivore defoliation due to parasitism by this hymenopteran parasitoid.     

CryIIIA toxin gene expression in transgenic rice confers resistance to rice water weevil

The rice water weevil (RWW), Lissorhoptrus oryzophilus Kuschel, is the most widely distributed and destructive early season insect pest of rice, Oryza sativa L. worldwide. The rice plants were transformed with cryIIIA insecticidal gene as well as with the bar gene coding phosphinothricin acetyltransferase. CryIIIA gene under the control of a modified RCg2 promoter drives the insect-toxic gene expression in roots and/or leaves. The cryIIIA gene was transferred into O. sativa L. cv. Nakdong by Agrobacterium-mediated transformation. Stable integration of the transgene was confirmed in putative transformed rice by Southern blot analysis. The expression of the cryIIIA toxin gene in the roots of transgenic rice plants was verified by RT-PCR and immunoblot analysis. Transgenic rice plants were also evaluated for resistance to natural infestations of the RWW under field conditions between 2007 and 2011. The transgenic Btt8R and Btt12R lines reduced the growth rate of RWW larvae and pupae populations compared with non-transgenic control plants by approximately 52 and 58 %, respectively. To further examine the efficacy of the RWW bioassay, we used pots and performed experiments in trays and under field conditions in 2012. The Btt12R line reduced the total populations of RWW larvae and pupae in trays and under field conditions by 56 and 45 %, respectively. The bioassay experiments conducted over 6 years, showed a significant reduction rate of RWW larvae and pupae populations demonstrating that the cryIIIA gene in transgenic rice confers resistance to RWW.     

The effects of feeding by the rice water weevil, Lissorhoptrus oryzophilus Kuschel, on the growth and yield components of rice, Oryza sativa

Abstract 1 The effects of feeding by larvae of the rice water weevil, Lissorhoptrus oryzophilus Kuschel, on the growth and yield components of rice, Oryza sativa, were evaluated using four varieties of rice, ‘Cocodrie’, ‘Cypress’, ‘Bengal’ and ‘XP1003’, over 2 years. 2 Both adults and larvae of L. oryzophilus feed on rice plants, but it is feeding by the larval stage that causes economic losses. Feeding by L. oryzophilus larvae resulted in extensive damage to root systems. Pruning of root systems resulted in a decrease in tiller number and shoot biomass of rice plants in the vegetative stage of growth. 3 Yield losses were due to a combination of decreases in panicle densities, numbers of grains per panicle, and grain weights. Decreases in panicle densities were a direct result of the reductions in tiller numbers. Reductions in numbers of grains per panicle and grain weights probably resulted from decreases in shoot biomass. 4 Injury by rice water weevil larvae is chronic. The tillering stage of rice suffered the majority of weevil damage, but the growth effects were not manifested until later.     

The influence of plant age on tolerance of rice to injury by the rice water weevil,Lissorhoptrus oryzophilus (Coleoptera: Curculionidae)

For most plant species, tolerance to many types of herbivory increases as plants age, but the applicability of this pattern to root herbivory has not been tested. Injury to roots of rice plants by larvae of the rice water weevil, Lissorhoptrus oryzophilus Kuschel, causes severe reductions in yields in the United States. It is generally thought that young rice plants, because their root systems are smaller, are less tolerant than older plants of root feeding by L. oryzophilus. Field experiments were conducted to test this hypothesis. Plots of rice (4.7 to 6.5 m2) were established and subjected to natural infestations of L. oryzophilus larvae. A soil insecticide was applied to plots at different times during the tillering phase of rice in order to manipulate the timing of weevil infestation. The impact of these treatments (timings of insecticide applications) was assessed by comparing relationships between yield loss and larval pressure for each treatment using analysis of covariance. Yield losses ranged from 13% to over 40% in plots not treated with insecticide. Patterns of yield losses from plots treated with insecticide at different times were best explained by the hypothesis that yield loss is determined both by the age of plants infested and by the size of larvae infesting plants. Young plants appear to be less tolerant than older plants, and feeding by large larvae appears to be more deleterious than feeding by smaller larvae. Management practices that delay infestation of rice by L. oryzophilus until plants are older may be an important component of management programmes for this pest.     

Wound-induced changes in tomato leaves and their effects on the feeding patterns of larval lepidoptera

Several studies have shown changes in the patterns of damage from feeding insects associated with changes in palatability and overall consumption as a result of wound-induced chemical changes in plants. This paper describes how the pattern of feeding damage made by the larvae of Spodoptera littoralis Boisd. (Lepidoptera: Noctuidae) on tomato is affected by changes in palatability of the leaves. Two sorts of responses to leaves from plants that had received prior damage were observed. Larvae offered a choice of leaves tended to take fewer meals on leaves from previously-wounded plants than on control leaves, frequently rejecting the former after sampling them. On wounded plants this rejection behaviour was associated with a shift in feeding site towards the base of the plant. However, starved larvae offered only a single excised leaf readily ate leaves from wounded plants but took shorter meals on these leaves than on controls. Although it was not directly tested it is possible that this difference in response reflected changes in food selectivity with a differing level of satiation. The results are considered in relation to the adaptive significance of the plant of changes in within-plant distributions of herbivore damage.     

Food preference and performance of the larvae of a specialist herbivore: variation among and within host-plant populations

Specialist herbivores are suggested to be unaffected by or attracted to the defense compounds of their host-plants, and can even prefer higher levels of certain chemicals. Abrostola asclepiadis is a specialist herbivore whose larvae feed on the leaves of Vincetoxicum hirundinaria, which contains toxic alkaloids and is unpalatable to most generalist herbivores. The food choice, leaf consumption and growth of A. asclepiadis larvae were studied to determine whether there is variation among and within host-plant populations in their suitability for this specialist herbivore. There was significant variation in food preference and leaf consumption among host-plant populations, but no differences were found in larval growth and feeding on different host-plant populations. A. asclepiadis larvae preferred host-plant populations with higher alkaloid concentrations, but did not consume more leaf material from plants originating from such populations in a no-choice experiment. There was also some variation in food preference of larvae among host-plant individuals belonging to the same population, suggesting that there was variability in leaf chemistry also within populations. Such variation in larval preference among host-plant genotypes and populations may create potential for coevolutionary dynamics in a spatial mosaic.     

Effect of arbuscular mycorrhizal fungi (Glomus intraradices) on the oviposition of rice water weevil (Lissorhoptrus oryzophilus)

Root-feeding insects are important drivers in ecosystems, and links between aboveground oviposition preference and belowground larval performance have been suggested. The root-colonizing arbuscular mycorrhizal fungi (AMF) play a central role in plant nutrition and are known to change host quality for root-feeding insects. However, it is not known if and how AMF affect the aboveground oviposition of insects whose offspring feed on roots. According to the preference–performance hypothesis, insect herbivores oviposit on plants that will maximize offspring performance. In a greenhouse experiment with rice (Oryza sativa), we investigated the effects of AMF (Glomus intraradices) on aboveground oviposition of rice water weevil (Lissorhoptrus oryzophilus), the larvae of which feed belowground on the roots. Oviposition (i.e., the numbers of eggs laid by weevil females in leaf sheaths) was enhanced when the plants were colonized by AMF. However, the leaf area consumed by adult weevils was not affected. Although AMF reduced plant biomass, it increased nitrogen (N) and phosphorus concentrations in leaves and N in roots. The results suggest that rice water weevil females are able to discriminate plants for oviposition depending on their mycorrhizal status. The discrimination is probably related to AMF-mediated changes in plant quality, i.e., the females choose to oviposit more on plants with higher nutrient concentrations to potentially optimize offspring performance. AMF-mediated change in plant host choice for chewing insect oviposition is a novel aspect of below- and aboveground interactions.     

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.     

The role of plant genotype,environment and gender in resistance to a specialist chrysomelid herbivore

Summary Patchiness in herbivore attack is a well-documented phenomenon. When neighboring plants suffer vastly different levels of attack, then one suspects genotypic differences among plants to be the underlying mechanism. In this study, I use common garden experiments in two natural, but divergent, habitats at the Cedar Creek Natural History Area in central Minnesota to determine the role of plant genotype, environment and gender in plant resistance to a specialist herbivore. Resistance was measured by larval survivorship and weight. Eight clones ofRhus glabra were selected and 12 equal-aged ramets were dug up and planted in two gardens (each garden received 6 ramets per clone). First instarBlepharida rhois (Coleoptera: Chrysomelidae) larvae of known parentage were transferred to ramets and censused every other day. At the end of the experiment, larvae were collected and weighed. Analysis of variance was used to determine the importance of plant genotype, environment and gender on larval mortality and weight. The experiment was repeated in its entirety one month later. Both plant genotype and environment significantly affected larval survivorship in the first run of the experiment. No interactions were significant. Results from the second run indicated marginally significant genotype and environment main effects, and a genotype by environment interaction in larval survivorship. There was a significant genotype by environment interaction in larval weight on the same run. In neither run did clone gender have significant affects on resistance.     

Movement and disappearance of mountain birch defoliators are influenced by the interactive effects of plant architecture and induced resistance

Abstract. 1. Changes in herbivore movement and feeding behaviour may determine the efficacy of induced plant resistance by affecting the location of damage within the foliage and by modifying the vulnerability of herbivores to predators. 2. Observations of larval feeding sites were used to test whether induced resistance increased the movement of free‐living Epirrita autumnata Borkh. (Lepidoptera, Geometridae) larvae feeding on mountain birch [Betula pubescens ssp. czerepanovii (Orlova) Hämet‐Ahti]. The amount of defoliation at different canopy parts was measured to test the associated changes in the spread of damage within the foliage. 3. The architectural complexity of trees was measured to test its association with the disappearance of larvae from their hosts. The underlying hypothesis was that the architectural traits of the host plant could affect disappearance by influencing the frequency of herbivores encountering predators. 4. Distance between the consecutive feeding positions, the number of leaves damaged, and consumption of long shoot leaves all increased in trees with induced resistance. 5. Disappearance of larvae depended on the architectural complexity of trees. The effect of complexity differed between defoliation treatments, and may depend on the activity and number of predators in relation to the canopy size. 6. Accordingly, this study suggested that the interactive effects of plant architecture, induced resistance, and herbivore behaviour can determine the performance of herbivores on their host plant.     

Feeding damage by larvae of the mustard leaf beetle deters conspecific females from oviposition and feeding

Herbivorous insects may be informed about the presence of competitors on the same host plant by a variety of cues. These cues can derive from either the competitor itself or the damaged plant. In the mustard leaf beetle Phaedon cochleariae (Coleoptera, Chrysomelidae), adults are known to be deterred from feeding and oviposition by the exocrine glandular secretion of conspecific co-occurring larvae. We hypothesised that the exocrine larval secretion released by feeding larvae may adsorb to the surface of Chinese cabbage leaves, and thus, convey the information about their former or actual presence. Further experiments tested the influence of leaves damaged by conspecific larvae, mechanically damaged leaves, larval frass and regurgitant on the oviposition and feeding behaviour of P. cochleariae. Finally, the effect of previous conspecific herbivory on larval development and larval host selection was assessed. Our results show that (epi)chrysomelidial, the major component of the exocrine secretion from P. cochleariae larvae, was detectable by GC-MS in surface extracts from leaves upon which larvae had fed. However, leaves exposed to volatiles of the larval secretion were not avoided by female P. cochleariae for feeding or oviposition. Thus, we conclude that secretion volatiles did not adsorb in sufficient amounts on the leaf surface to display deterrent activity towards adults. By contrast, gravid females avoided to feed and lay their eggs on leaves damaged by second-instar larvae for three days when compared to undamaged leaves. Mechanical damage of leaves and treatment of artificially damaged leaves with larval frass or regurgitant did not affect oviposition and feeding of P. cochleariae. Since no adverse effects of previous herbivory on larval development were detected, we suggest that female P. cochleariae avoid Chinese cabbage leaves damaged by feeding larvae for other reasons than escape from competition or avoidance of direct negative effects that result from consuming induced plant material.     

Experimental evaluation of the impacts of two ant species on banana weevil in Uganda

The banana weevil, Cosmopolites sordidus (Germar), is an important pest of bananas. Predatory ants are increasingly being viewed as possible biological control agents of this pest because they are capable of entering banana plants and soil in search of prey. We studied ant predation on banana weevil in Uganda in crop residues and live plants in both laboratory and field experiments. Field studies with live plants used chemical ant exclusion in some plots and ant enhancement via colony transfer in others to measure effects of Pheidole sp. 2 and Odontomachus troglodytes Santschi on plant damage and densities of immature banana weevils.In crop residues, an important pest breeding site, twice as many larvae were removed from ant-enhanced plots as in control plots. In young (2 month) potted suckers held in shade houses, ant ability to reduce densities of banana weevil life stages varied with the weevil inoculation rate. At the lowest density (2 female weevils per pot), densities of eggs, larvae, and pupae were reduced by ants. At higher rates there was no effect. In older suckers (5–11 months) grown in larger containers, banana weevil densities were not affected by ants, but damage levels were reduced. In a field trial lasting a full crop cycle (30 months), we found that the ants tested reduced the density of banana weevil eggs in suckers during the crop, but did not affect larval densities in the sampled suckers. However, most larvae occur in the main banana plants, rather than associated suckers. Nevertheless, levels of damage in mature plants at harvest did not differ between Amdro-treated and ant-enhanced plots, suggesting the ant species studied were not able to provide economic control of banana weevil under our test conditions.     

<Emphasis Type="Italic">Piriformospora indica</Emphasis> and <Emphasis Type="Italic">Sebacina vermifera</Emphasis> increase growth performance at the expense of herbivore resistance in <Emphasis Type="Italic">Nicotiana attenuata</Emphasis>

A Sebacinales species was recovered from a clone library made from a pooled rhizosphere sample of Nicotiana attenuata plants from 14 native populations. Axenic cultures of the related species, Piriformospora indica and Sebacina vermifera, were used to examine their effects on plant performance. Inoculation of N. attenuata seeds with either fungus species stimulated seed germination and increased growth and stalk elongation. S. vermifera inoculated plants flowered earlier, produced more flowers and matured more seed capsules than did non-inoculated plants. Jasmonate treatment during rosette-stage growth, which slows growth and elicits herbivore resistance traits, erased differences in vegetative, but not reproductive performance resulting from S. vermifera inoculation. Total nitrogen and phosphorous contents did not differ between inoculated and control plants, suggesting that the performance benefits of fungal inoculation did not result from improvements in nutritional status. Since the expression of trypsin proteinase inhibitors (TPI), defensive proteins which confer resistance to attack from Manduca sexta larvae, incur significant growth and fitness costs for the plant, we examined the effect of S. vermifera inoculation on herbivore resistance and TPI activity. After 10 days of feeding on S. vermifera-inoculated plants, larval mass was 46% higher and TPI activity was 48% lower than that on non-inoculated plants. These results suggest that Sebacina spp. may interfere with defense signaling and allow plants to increase growth rates at the expense of herbivore resistance mediated by TPIs.     

Fine‐tuning of defences and counter‐defences in a specialised plant–herbivore system

1. The plant–herbivore arms race has been postulated to be a major driver for generating biological and biochemical diversity on Earth. Herbivore feeding is reduced by the production of chemical and physical barriers, but increases plant resistance against subsequent attack. Accordingly, specialisation is predicted to be an outcome of herbivores being able to circumvent plant‐induced defences. 2. Using a specialised plant–herbivore system, in which adult chrysomelid beetles (Chrysochus auratus) feed on leaves and larvae feed on roots of dogbane (Apocynum spp.), this study investigated whether root latex and cardenolides are effective against the soil‐dwelling larvae, and whether such defences could be circumvented by the herbivore. 3. Across two Apocynum species, C. auratus larvae were not affected by latex production or cardenolide amounts and diversity. By contrast, cardenolide apolarity was detrimental to larval growth. Yet larval feeding decreased average root cardenolide apolarity in A. cannabinum and larvae performed better on those plants. Finally, above‐ground induction rendered the plants more toxic by increasing root cardenolide apolarity and maintaining it, even during subsequent larval herbivory. 4. Therefore, the intimate relationship and interaction between Chrysochus and Apocynum are maintained by a delicate balance of herbivore manipulation and plant chemical induction.     

In vitro insect-feeding bioassay to determine the resistance of transgenic rice plants transformed with insect resistance genes against striped stem borer (Chilo suppressalis)

Summary To determine the degree of insect resistance in transgenic plants, different bioassays are used which typically use either whole plant or small pieces of leaves or stems of transgenic plants, following culture under greenhouse conditions. An in vitro insect-feeding bioassay is presented which permits the infestation of transgenic plantlets with newly hatched larvae from the striped stem borer. The bioassay consists of the germination of rice seeds in vitro using Murashige and Skoog medium in test tubes, and then infestation of each 3–4 cm long seedling with one neonate larva obtained from surfacesterilized eggs of Chilo suppressalis. The infested in vitro plantlets are kept in culture rooms at 25°C for several days and then the seedling damage and the growth of the larvae are analyzed. Senia (japonica variety) homozygous transgenic rice plants were used for these experiments. The plants were transformed with either the cry1B or the maize proteinase inhibitor (mpi) genes. Both genes confer resistance to Chilo suppressalis. With non-transformed plants the larvae grew and developed normally, feeding on the small rice plantlets. In contrast, with cry1B plants, the neonate larvae died during the first days of the infestation. These plantlets recovered completely and developed similarly to the non-infested control plants. With transgenic plants transformed with the mpi gene, the neonate larvae did not die but grew more slowly compared with the controls. Thus, this in vitro insect-feeding bioassay is a rapid and easy method to detect the resistance of cry and mpi transgenic plants to stem borers such as Chilo suppressalis.