Insecticidal Bt Research Articles

Critical Facets of European Corn Borer Adult Movement Ecology Relevant to Mitigating Field Resistance to Bt-Corn.

The European corn borer (Ostrinia nubilalis, Hübner) has been managed successfully in North America since 1996 with transgenic Bt-corn. However, field-evolved resistance to all four available insecticidal Bt proteins has been detected in four provinces of Canada since 2018. Evidence suggests resistance may be spreading and evolving independently in scattered hotspots. Evolution and spread of resistance are functions of gene flow, and therefore dispersal, so design of effective resistance management and mitigation plans must take insect movement into account. Recent advances in characterizing European corn borer movement ecology have revealed a number of surprises, chief among them that a large percentage of adults disperse from the natal field via true migratory behavior, most before mating. This undermines a number of common key assumptions about adult behavior, patterns of movement, and gene flow, and stresses the need to reassess how ecological data are interpreted and how movement in models should be parameterized. While many questions remain concerning adult European corn borer movement ecology, the information currently available is coherent enough to construct a generalized framework useful for estimating the spatial scale required to implement possible Bt-resistance prevention, remediation, and mitigation strategies, and to assess their realistic chances of success.

ABC transporter subfamily B1 as a susceptibility determinant of Bombyx mori larvae to Cry1Ba, Cry1Ia and Cry9Da toxins

ATP binding cassette (ABC) transporters are a diverse family of transmembrane proteins. Specific subfamily members expressed in the lepidopteran midgut can act as susceptibility determinants for several insecticidal Bt Cry proteins. However, the susceptibility determinants to many Cry toxins still remain unclear. Therefore, we knocked out a series of ABC transporters that are highly expressed in the midgut of Bombyx mori larvae by transcription activator-like effector nuclease (TALEN)-mediated gene editing, and the lineages that became resistant to Cry toxins were searched by toxin overlay bioassay. As a result, the B. mori ABC transporter subfamily B1 (BmABCB1) knockout lineage showed 19.17-fold resistance to Cry1Ba, 876.2-fold resistance to Cry1Ia, and 29.1-fold resistance to Cry9Da, suggesting that BmABCB1 is the determinant of susceptibility to these toxins. BmABCC2 and BmABCC3 have been shown to be susceptibility determinants based on their function as receptors. Therefore, we next heterologously expressed these ABC transporters in HEK293T cells and performed a cell swelling assay to examine whether these molecules could exert receptor functions. As a result, BmABCB1-expressing cells showed swelling response to Cry1Ia and Cry9Da, and cells expressing PxABCB1, which is the Plutella xylostella ortholog of BmABCB1, showed swelling for Cry1Ba, suggesting that ABCB1 is a susceptibility determinant by functioning as a receptor to these toxins. Furthermore, in order to clarify how high binding affinity is based on receptor function, we performed surface plasmon resonance analysis and found that each KD of Cry1Ba, Cry1Ia, and Cry9Da to BmABCB1 were 7.69 × 10−8 M, 2.19 × 10−9 M, and 4.17 × 10−6 M respectively.

Is ‘pupae busting’ or destroying overwintering pupae of Helicoverpa spp. (Lepidoptera: Noctuidae) still relevant today in Australian Bt cotton?

AbstractTransgenic Bt cotton was developed to control lepidopteran pests like the cotton bollworm, Helicoverpa armigera. However, there was concern that H. armigera would develop resistance to Bt cotton as this species had developed resistance to many insecticides and Bt toxins. To counter resistance, the cotton industry developed a resistance management plan (RMP) that included techniques to block resistant genes surviving from one season to the next (seasonal quarantining). One such technique is pupae busting, where cotton fields are cultivated after harvest, destroying potentially resistant pupating Helicoverpa spp. While pupae busting was important when there was only one insecticidal gene in Bt cotton, is it still relevant now Bt cotton has three insecticidal Bt genes? To address this question, we reviewed the development of pupae busting as a tool and its role in the current RMP. This included examining the ecology and behavioural characteristics of Helicoverpa spp. that impact on pupae busting efficacy (e.g., diapause, pupal mortality and pupae depth); the effect of soil type and different tillage techniques on pupae busting efficacy; and pupae busting within the context of Australia's current cotton farming system. We also looked at alternative forms of seasonal quarantining, such as using bisexual attract‐and‐kill techniques against adults. We confirmed that soil for pupae busting needs to be checked for moisture, which ideally should be less than the soil plastic limit. Comparisons between reports indicated that under good conditions, ‘go‐devils’ and chisel ploughs were excellent pupae busters. While a bisexual attract‐and‐kill strategy of late season moths has a place within the industry, pupae busting is still the best method in seasonal quarantining and has a good fit within the modern cotton industry, particularly given differences in the biology and ecology of H. armigera and H. punctigera, and the presence of dominant resistance to Bt toxins by H. armigera in China.

Susceptibility and diagnostic concentration for Bacillus thuringiensis toxins and newer chemical insecticides in Spodoptera frugiperda (Lepidoptera: Noctuidae) from China.

The fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), is a cosmopolitan pest that exploits more than 350 host plants, including economically important crops such as corn, cotton and rice. Control of S. frugiperda largely relies on transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) and spraying synthetic insecticides. Here, we established the susceptibility and diagnostic concentration for 2 Bt toxins and 5 newer insecticides in invasive populations of S. frugiperda from southeastern China. Concentrations causing 50% mortality (LC50) in ten field populations sampled in 2022 ranged from 2.13 to 19.29 and 22.43 to 71.12 ng/cm2 for Cry1Fa and Vip3Aa, and 0.83 to 5.30, 2.83 to 9.94, 0.04 to 0.23, 4.59 to 8.40, and 1.49 to 6.79 mg/liter for chlorantraniliprole, chlorfenapyr, emamectin benzoate, indoxacarb, and spinosad, respectively. Relative to the susceptible strain YJ-19, the largest resistance ratio in the field populations was 5.1, 1.6, 6.2, 3.9, 4.6, 2.2, and 3.6 for Cry1Fa, Vip3Aa, chlorantraniliprole, chlorfenapyr, emamectin benzoate, indoxacarb, and spinosad, respectively, indicating that the field populations were generally susceptible to these Bt toxins and insecticides. Based on the pooled response of the field populations, the diagnostic concentration for resistance monitoring, estimated as ca. twice the LC99, was 400 and 1,500 ng/cm2 for Cry1Fa and Vip3Aa, and 2, 40, 60, 60, and 100 mg/liter for emamectin benzoate, chlorantraniliprole, chlorfenapyr, spinosad, and indoxacarb, respectively. These results provide useful information for monitoring resistance to key Bt toxins and insecticides for the control of S. frugiperda in China.

Enhancing insecticidal efficacy of Bacillus thuringiensis Cry1Ab through pH-sensitive encapsulation.

Nanotechnology is a promising way to enhance the stability of Bacillus thuringiensis (Bt) insecticidal proteins under environmental conditions. In this work, two emulsions were prepared through the Pickering emulsion technique, stabilized by Cu2+-SQDs/S-CN nanocomposites and by GO nanosheets. In addition, a pH-sensitive polymer was incorporated into these emulsions, allowing the Bt protein, Cry1Ab, to be released in an alkaline pH environment, as it occurs in the lepidopteran pests' gut. The effectiveness of these two nanomaterials in protecting Cry1Ab from degradation, and therefore enhancing its pesticidal activity, was assessed by exposing samples of the purified unprotected protein and encapsulated protein to high-intensity UV light and 40°C temperature treatments. The UV treatment results were evaluated using SDS-PAGE analysis and pointed out that Cry1Ab could be structurally protected by the emulsions. The bioassays with first instar larvae of the lepidopteran pest Ostrinia nubilalis confirm the nanomaterial protection to UV and temperature treatments, i.e., decreasing about half the degradation rate and increasing up to 12-fold the residual activity after UV treatment. Our results indicate that encapsulation could be an effective strategy to improve the effectiveness of Cry1Ab under environmental conditions. KEY POINTS: • Pickering emulsions are effective for solubilized Cry1Ab encapsulation. • Structural and toxicity Cry1Ab properties are enhanced by pH-sensitive encapsulation. • Cu2+-SQDs/S-CN and GO nanomaterials improve the efficacy of Bt insecticides.

Effects of Bacillus thuringiensis on biological parameters of Tetrastichus howardi parasitizing Bt-resistant pupa of Spodoptera frugiperda

The endoparasitoid wasp, Tetrastichus howardi Olliff, is used for insect pest control in agricultural systems. The toxic effects of proteins produced by Bacillus thuringiensis Berliner (Bt) were evaluated over two generations of T. howardi parasitizing Bt-resistant Spodoptera frugiperda. Lower survivorship was detected in both generations of parasitoid wasps of both sexes that developed on S. frugiperda larvae resistant to Bt. Deleterious effects including disrupted host-searching behavior and lower performance were found in T. howardi individuals that emerged from Bt-resistant S. frugiperda. Also, Bt caused repellency and reduced immature parasitoid wasp production in T. howardi. The findings revealed that Bt causes side-effects on T. howardi which has implications for assessing the risks of Bt insecticides in biological control programs for sustainable crop production.

Managing Spodoptera Species (Lepidoptera: Noctuidae) Found in Brazilian Soybean Fields with Bt Soybean and Insecticides

Genetically modified (GM) soybeans expressing Cry1A.105/Cry2Ab2/Cry1Ac (event MON 87701 × MON 89788 × MON 87751 × MON 87708) and Cry1Ac (event MON 87701 × MON 89788) from Bacillus thuringiensis Berliner (Bt) are valuable technologies for managing key lepidopteran pests of soybean in South America, but do not provide stand-alone protection against Spodoptera species. Here, we evaluated the use of these Bt soybeans and their integration with insecticides for managing Spodoptera species. Cry1A.105/Cry2Ab2/Cry1Ac soybean provided reasonable levels of protection against S. cosmioides, S. albula, and S. eridania. However, S. frugiperda had higher survival on this Bt soybean, and Cry1Ac soybean showed low lethality against all species evaluated. Spodoptera larvae that survived on Bt and non-Bt soybean showed comparable susceptibility to flubendiamide and thiodicarb in diet-overlay bioassays. Regardless of soybean plant type, the field doses of flubendiamide and thiodicarb were effective in controlling surviving Spodoptera larvae. We conclude that Cry1A.105/Cry2Ab2/Cry1Ac soybean is effective in controlling S. cosmioides and S. albula, and also has reasonable control of S. eridania, but not S. frugiperda. Cry1Ac soybean provided poor control of all Spodoptera species. Nonetheless, Spodoptera larvae surviving on both Bt and non-Bt soybean were controlled by flubendiamide and thiodicarb. Thus, integrated control tactics would provide effective management of Spodoptera species in Bt soybean fields in South America.

ARTP and NTG compound mutations improved Cry protein production and virulence of Bacillus thuringiensis X023.

A high production mutated strain Bacillus thuringiensis X023PN (BtX023PN) was screened from the wild strain Bacillus thuringiensis X023 (BtX023) after atmospheric and room temperature plasma (ARTP) and nitrosoguanidine (NTG) mutation. BtX023PN grows faster than the wild strain, and its lysis of mother cell was 6h ahead BtX023, but the ability of sporulation was significantly reduced. Bioassay indicated that compared with the wild type strain, the virulence of BtX023PN against Plutella xylostella (P. xylostella) and Mythimna seperata (M. seperata) increased to 2.33-fold and 2.13-fold respectively. qRT-PCR and SDS-PAGE demonstrated that the production of Cry1Ac increased by 61%. Resequence indicated that the mutated sites enriched on the key carbohydrate metabolism and amino acid metabolism. This study provides a new strain resource for the development of Bt insecticides and a feasible technical strategy for the breeding of Bt. KEY POINTS: • Atmospheric and room temperature plasma used in breeding of Bacillus thuringiensis. • Less stationary phase time with more ICP production. • Semi-lethal concentration against Plutella xylostella reduced by about 57.

Bacillus cereus in Packaging Material: Molecular and Phenotypical Diversity Revealed.

The Bacillus cereus group has been isolated from soils, water, plants and numerous food products. These species can produce a variety of toxins including several enterotoxins [non-hemolytic enterotoxin (Nhe), hemolysin BL (Hbl), cytotoxin K, and enterotoxin FM], the emetic toxin cereulide and insecticidal Bt toxins. This is the first study evaluating the presence of B. cereus in packaging material. Among 75 different isolates, four phylogenetic groups were detected (II, III, IV, and VI), of which the groups III and IV were the most abundant with 46.7 and 41.3%, respectively. One isolate was affiliated to psychrotolerant group VI. Growth experiments showed a mesophilic predominance. Based on PCR analysis, nhe genes were detectable in 100% of the isolates, while hbl genes were only found in 50.7%. The cereulide encoding gene was found in four out of 75 isolates, no isolate carried a crystal toxin gene. In total, thirteen different toxin gene profiles were identified. We showed that a variety of B. cereus group strains can be found in packaging material. Here, this variety lies in the presence of four phylogenetic groups, thirteen toxin gene profiles, and different growth temperatures. The results suggest that packaging material does not contain significant amounts of highly virulent strains, and the low number of cereulide producing strains is in accordance with other results.

Plant Polysaccharides Modulate Biofilm Formation and Insecticidal Activities of Bacillus thuringiensis Strains.

After the biological pesticide Bacillus thuringiensis (Bt) is applied to the field, it has to remain on the surface of plants to have the insecticidal activities against insect pests. Bt can form biofilms on the surface of vegetable leaves, which were rich in polysaccharides. However, the relationship between polysaccharides of the leaves and the biofilm formation as well as the insecticidal activities of Bt is still unknown. Herein, this study focused on the effects of plant polysaccharides pectin and xylan on biofilm formation and the insecticidal activities of Bt strains. By adding pectin, there were 88 Bt strains with strong biofilm formation, 69 strains with weak biofilm formation, and 13 strains without biofilm formation. When xylan was added, 13 Bt strains formed strong biofilms, 98 strains formed weak biofilms, and 59 strains did not form biofilms. This indicated that two plant polysaccharides, especially pectin, modulate the biofilm formation of Bt strains. The ability of pectin to induce biofilm formation was not related to Bt serotypes. Pectin promoted the biofilms formed by Bt cells in the logarithmic growth phase and lysis phase at the air–liquid interface, while it inhibited the biofilms formed by Bt cells in the sporangial phase at the air–liquid interface. The dosage of pectin was positively correlated with the yield of biofilms formed by Bt cells in the logarithmic growth phase or lysis phase at the solid–liquid interfaces. Pectin did not change the free-living growth and the cell motility of Bt strains. Pectin can improve the biocontrol activities of the spore–insecticidal crystal protein mixture of Bt and BtK commercial insecticides, as well as the biofilms formed by the logarithmic growth phase or lysis phase of Bt cells. Our findings confirmed that plant polysaccharides modulate biofilm formation and insecticidal activities of Bt strains and built a foundation for the construction of biofilm-type Bt biopesticides.

Geographic Monitoring of Insecticide Resistance Mutations in Native and Invasive Populations of the Fall Armyworm.

Simple SummaryThe moth fall armyworm (Spodoptera frugiperda) is a major agricultural pest insect damaging a wide range of crops, especially corn. Field evolved resistance against Bacillus thuringiensis (Bt) toxins and synthetic insecticides has been repeatedly reported. While the fall armyworm is native to the Americas, its biological invasion was first reported from West Africa in 2016. Since then, this pest has been detected across sub-Saharan and North Africa, Asia, and Oceania. Here, we examine the geographical distribution of mutations causing resistance against Bt or synthetic insecticides to test if the invasion was accompanied by the spread of resistance mutations using 177 individuals collected from 12 geographic populations including North and South America, West and East Africa, India, and China. We observed that Bt resistance mutations generated in Puerto Rico or Brazil were found only from their native populations, while invasive populations had higher copy numbers of cytochrome P450 genes and higher proportions of resistance mutations at AChE, which are known to cause resistance against synthetic insecticides. This result explains the susceptibility to Bt insecticides and the resistance against synthetic insecticides in invasive Chinese populations. This information will be helpful in investigating the cause and consequence associated with insecticide resistance.Field evolved resistance to insecticides is one of the main challenges in pest control. The fall armyworm (FAW) is a lepidopteran pest species causing severe crop losses, especially corn. While native to the Americas, the presence of FAW was confirmed in West Africa in 2016. Since then, the FAW has been detected in over 70 countries covering sub-Saharan Africa, the Middle East, North Africa, South Asia, Southeast Asia, and Oceania. In this study, we tested whether this invasion was accompanied by the spread of resistance mutations from native to invasive areas. We observed that mutations causing Bt resistance at ABCC2 genes were observed only in native populations where the mutations were initially reported. Invasive populations were found to have higher gene numbers of cytochrome P450 genes than native populations and a higher proportion of multiple resistance mutations at acetylcholinesterase genes, supporting strong selective pressure for resistance against synthetic insecticides. This result explains the susceptibility to Bt insecticides and resistance to various synthetic insecticides in Chinese populations. These results highlight the necessity of regular and standardized monitoring of insecticide resistance in invasive populations using both genomic approaches and bioassay experiments.

Cryo-EM structures of an insecticidal Bt toxin reveal its mechanism of action on the membrane

Insect pests are a major cause of crop losses worldwide, with an estimated economic cost of $470 billion annually. Biotechnological tools have been introduced to control such insects without the need for chemical pesticides; for instance, the development of transgenic plants harbouring genes encoding insecticidal proteins. The Vip3 (vegetative insecticidal protein 3) family proteins from Bacillus thuringiensis convey toxicity to species within the Lepidoptera, and have wide potential applications in commercial agriculture. Vip3 proteins are proposed to exert their insecticidal activity through pore formation, though to date there is no mechanistic description of how this occurs on the membrane. Here we present cryo-EM structures of a Vip3 family toxin in both inactive and activated forms in conjunction with structural and functional data on toxin–membrane interactions. Together these data demonstrate that activated Vip3Bc1 complex is able to insert into membranes in a highly efficient manner, indicating that receptor binding is the likely driver of Vip3 specificity.

Susceptibility of the Egyptian Cotton Leafworm, Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae) to Entomocidal Crystal Proteins Cry1Ac and Cry 2Ab Baseline Responses

The present study maybe is the first attempt in Egypt to evaluate the effect of Entomocidal proteins (Cry toxins) from Bacillus thuringiensis (Bt)Cry 1Ac and Cry 2Ab againstthe Egyptian cotton leafworm Spodoptera littoralis. The cotton leafworm one of the important Lepidopteran species is the most important pests in cotton fields. It can be controlled using Genetically Modified (GM) crops expressing insecticidal Bt proteins like Bt cotton and Bt corn. The long-term success of this technology demands pest resistance management. Important information for the successful management of resistance is the baseline susceptibility of the different target pests to the different Bt proteins. The data on baseline susceptibility should enable risk assessors and managers to assess whether GM cotton produces a Bt protein in a high-dose to specific target organisms and resistance has evolved during the commercial cultivation of this GM cotton event. During this study, the toxicity of Cry 1Ac and Cry 2Ab from the bacterium Bacillus thuringiensis to larvae of S. littoralis was assessed using first instar larvae with diet incorporation bioassays. Larvae reared (4 generations) from 2 populations of S. littoralis collected from major cotton-growing regions of Sohag and Assuit Governorates were tested. The LC50 for the Assuit strain was 140.11 and 88.54 µg/ml forCry 1Ac and Cry 2Ab, respectively. While, the LC50 for the Sohag strain was 410.75 and 37.62 µg/ml for Cry 1Ac and Cry 2Ab, respectively. On the other hand, the laboratory strain was more sensitive for the same Cry’s (Cry 1Ac and Cry 2Ab) the LC50 was13.40 and 0.23 µg/ml, respectively. The lethal concentration (LC50) against S. littoralis for 2nd and 4th instars larvae when using the B. thuringiensis as spores (commercial product) was 1.16 and 2.49 for 2nd and 4th instars, respectively. This study is very important as a preliminary step and baseline ahead of the commercial cultivation and production of Genetically Modified cotton (Bt cotton).

Fused: a promising molecular target for an RNAi-based strategy to manage Bt resistance in Plutella xylostella (L.)

Bacillus thuringiensis (Bt), a naturally occurring entomopathogenic soil bacterium, has been the active ingredient of sprayable Bt biopesticides for over a century. Insecticidal Bt Cry proteins are particularly well suited for use as plant-incorporated protectants in transgenic crops (Bt crops) due to their specificity against insect pests and safety for non-target organisms. The sustainability of these Bt products, however, has been challenged by the development of resistance in the field. RNA interference (RNAi), a species-specific control alternative that has been deregulated and commercialized in the USA and Canada, provides a new mode of action to complement the existing Bt products. Based on our preliminary research, we hypothesized that pyramiding/integrating Bt with RNAi can address/manage resistance issues related to Bt traits. To examine this overarching hypothesis, we (1) cloned and characterized a serine/threonine kinase gene (fused) of the Hedgehog (Hh) signaling pathway in the diamondback moth, Plutella xylostella (L.), a global superpest; (2) profiled Pxfused expression in Bt-resistant P. xylostella strains; and (3) investigated the involvement of Pxfused in Bt Cry1Ac resistance in P. xylostella. Pxfused expression was elevated ubiquitously in all Bt-resistant strains, and silencing of Pxfused led to larval and pupal mortality in both Cry1Ac-susceptible and -resistant strains, suggesting that Pxfused is a potential target for RNAi-based resistance management. Taken together, our results not only identify a molecular target to control a devastating lepidopteran pest, but also shed light on a novel resistance management strategy through the integration of two biotechnological techniques with distinct modes of action.

Mechanisms of Resistance to Insecticidal Proteins from Bacillus thuringiensis.

Insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) are used in sprayable formulations or produced in transgenic crops as the most successful alternatives to synthetic pesticides. The most relevant threat to sustainability of Bt insecticidal proteins (toxins) is the evolution of resistance in target pests. To date, high-level resistance to Bt sprays has been limited to one species in the field and another in commercial greenhouses. In contrast, there are currently seven lepidopteran and one coleopteran species that have evolved practical resistance to transgenic plants producing insecticidal Bt proteins. In this article, we present a review of the current knowledge on mechanisms of resistance to Bt toxins, with emphasis on key resistance genes and field-evolved resistance, to support improvement of Bt technology and its sustainability.

Needs Assessment for Corn Insect Pest Management in Iowa

AbstractCorn, Zea mays L. (Cyperales: Poaceae), production is essential to Iowa’s economy and provides food, feed, and fuel beyond the state. Farmers face a number of challenges in corn production, including insect pests, and understanding farmers’ perceptions and concerns is critical for extension personnel to provide adequate and relevant programming. The Iowa State University Agriculture and Natural Resources Crops Team conducted a survey of Iowa farmers and agribusiness professionals during the 2019–2020 extension season. The goal was to understand farmer perceptions of corn pests and whether farmers experienced challenges with resistance to Bt traits or insecticides. Additionally, we aimed to realize farmers’ confidence in developing resistance management plans and discussing those needs with their input suppliers and advisors. Our survey revealed that northern corn rootworm, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae), was the pest of greatest concern across the state. People were generally more concerned about belowground Bt trait performance than aboveground traits, though only 12% of respondents reported Bt trait failure for corn rootworms. In general, people were not concerned about the performance of seed treatments or soil- or foliar-applied insecticides. Farmers generally felt very confident in developing a resistance management plan and discussing their needs with their advisors. The results of this survey will be used to target extension programming throughout the state and increase confidence for farmers and agribusiness employees to discuss resistance management plans together.

Inseticidas nas sementes e a entomotoxina cry 1 ac protegem a soja contra a lagarta-do-velho-mundo

The Helicoverpa armigera caterpillar occurs from the emergence of the plants to the reproductive stage of the soybean crop, which can cause significant losses.As an initial pest, both neonate and more developed caterpillars can be found, the latter from the previous crop.The use of Bt cultivars and insecticides in seed treatment are options to pest managementat the beginning of soybean development, but there are doubtsabout the efficacy of these control methods. Thus, the objective was to evaluate the protective effect of insecticides applied to seeds and Cry 1Ac protein, in relation to the attack of the H. armigeracaterpillar, as an initial pest in soybean plants.Experiments were conducted in the laboratory, using a completely randomized design, with six treatments:four insecticides related to seeds and control (without insecticide), in conventional cultivar soybeans (Target BMX), and the Cry 1Ac protein in soybeans cultivate Bt (DM 5958 IPRO), without insecticide. Our results confirm that under laboratory conditions the insecticides cyantraniliprole (60 g ai / 100 kg of seeds) and chlorantraniliprole (62.5 g ai / 100 kg of seeds) protect the soybean against the injuries of H. armigera3º-4ºínstars, in cotyledons and unifoliolate and trifoliolate leaves, at 5 days after the emergenceof the plants. The protein Cry 1Acprotects the soybean plants from this type of injury caused byboth3rd-4thand 5th-6thínstarscaterpillars, at 5 and 10 days after emergence.

Development of diamondback moth resistant transgenic cabbage and cauliflower by stacking Cry1B and Cry1CBt genes

Cabbage and cauliflower are extensively cultivated Cole vegetables all around the world. These are highly prone to the most notorious insect pest Plutella xylostella (diamondback moth), causing huge yield losses to the growers. Conventional breeding for host plant resistance against this pest is limited due to lack of resistance source in the available germplasm. The chemical control measures are cumbersome, costly, health hazardous and environment unfriendly. Therefore, using quaternary gene pool for incorporation of resistance genes from unrelated sources such as bacterial derived insecticidal Bt toxins through genetic engineering is a viable option. Genetic manipulation of plants through Agrobacterium-mediated transformation is limited by a multitude of factors resulting in poor transformation efficiency. Hence, optimization of plant transformation method suitable for a particular crop is highly essential. Here in we report the optimization of plant transformation protocol for cabbage and cauliflower by the manipulation of various key parameters, including the type of explants, Agrobacterium strains, bacterial cell density, age of the explant, preculture period, media formulations etc. The binary vector pPIPRA560 containing stacked Bt genes (Cry1B/Cry1C) and npt II/Basta as selectable marker was chosen for plant transformation. As a proof of concept, initially the effectiveness of the Bt genes against diamondback moth was validated in transgenic Arabidopsis. Insect bioassay revealed 100% mortality within 48 h of feeding. Based on the exciting results, the same construct was used for transforming Indian popular cultivars of cabbage (‘Golden Acre’) and cauliflower (‘Pusa Meghna’ and ‘Pusa Snowball K1’). The presence of the Bt genes in the transgenic lines was confirmed by PCR and expression through RT-qPCR. Insect bioassay revealed 100% mortality in some of the plants tested so far.

Effects of Pest Management Practices on Soil Nematode Abundance, Diversity, Metabolic Footprint and Community Composition Under Paddy Rice Fields.

The wide-scale adoption of transgenic crops has aroused public concern towards potential impacts to the ecological services of soil fauna, such as soil nematodes. However, few studies has examined whether the cultivation of transgenic rice would pose greater threats to soil nematode community and associated ecological functions than insecticides application. Moreover, what are determinants of soil nematode community in paddy fields remains unclear. During a 3-year field study, rhizosphere soil samples of transgenic-Bt rice, its counterpart non-Bt parental rice and not-Bt rice with insecticides application were taken at four times in the rice developmental cycle using a random block design with three replications for each treatment. We hypothesized that the effects of pest management practice on soil nematode abundance and metabolic footprint change with trophic group and sampling time. We also predicted there were significant differences in structure and composition of soil nematode community across the three treatments examined and sampling times. In agreement with our expectation, the effects of pest management practice on nematode abundance and metabolic footprints depend on trophic group and sampling time. However, pest management practice exerted no apparent effect on nematode diversity and community composition. Soil nutrient availability and C:N molar ratio are the primary regulating factor of soil nematode community in rice paddy fields. In conclusion, our findings implied that changes in abundance, diversity, metabolic footprints associated with the crop growth stage overweighed the application of Bt rice and insecticides. The cultivation of Bt rice Huahui-1 exerted no measurable adverse effect on soil nematode community in rhizosphere soil over 3 years of rice cropping.

Association of Insect-Derived Ear Injury With Yield and Aflatoxin of Maize Hybrids Varying in Bt Transgenes.

Environmental factors have been associated with the production of aflatoxin in maize, Zea mays L., and it is inconclusive whether transgenic, Bacillus thuringiensis (Bt), maize has an impact on aflatoxin accumulation. Maize hybrids differing in transgenes were planted in two locations from 2014 to 2017. Yield, aflatoxin, and ear injury caused by corn earworm, Helicoverpa zea (Boddie), and fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), were measured across three groups of hybrids differing in transgenes including near-isogenic hybrids, and water-stressed conditions. The hybrid groups consisted of non-Bt hybrids with no Bt transgenes, a second group with one or more Cry-Bt transgenes, and the third group with vegetative insecticidal Bt protein and Cry-Bt transgenes (Cry/Vip-Bt). Across the six data sets derived from 11 experiments, the Cry-Bt and Cry/Vip-Bt hybrids had less ear injury and aflatoxin on average than non-Bt hybrids. The effects of ear injury on yield and aflatoxin were more prominent and consistent in Corpus Christi, TX, where hybrids experienced more water-limited conditions than in College Station, TX. The trend of increased aflatoxin among hybrids with increased ear injury was further resolved when looking at Cry-Bt and Cry/Vip-Bt isogenic hybrids in Corpus Christi. The results supported that the maize hybrids with the inclusion of Cry-Bt and Cry/Vip-Bt transgenes warrant further investigation in an integrated approach to insect and aflatoxin management in sub-tropical rain-fed maize production regions. Research outcomes may be improved by focusing on areas prone to water-stress and by using hybrids with similar genetic backgrounds.