Showing papers on "Bacillus thuringiensis published in 2010"

An ABC transporter mutation is correlated with insect resistance to Bacillus thuringiensis Cry1Ac toxin.

Abstract: Transgenic crops producing insecticidal toxins from Bacillus thuringiensis (Bt) are commercially successful in reducing pest damage, yet knowledge of resistance mechanisms that threaten their sustainability is incomplete. Insect resistance to the pore-forming Cry1Ac toxin is correlated with the loss of high-affinity, irreversible binding to the mid-gut membrane, but the genetic factors responsible for this change have been elusive. Mutations in a 12-cadherin-domain protein confer some Cry1Ac resistance but do not block this toxin binding in in vitro assays. We sought to identify mutations in other genes that might be responsible for the loss of binding. We employed a map-based cloning approach using a series of backcrosses with 1,060 progeny to identify a resistance gene in the cotton pest Heliothis virescens that segregated independently from the cadherin mutation. We found an inactivating mutation of the ABC transporter ABCC2 that is genetically linked to Cry1Ac resistance and is correlated with loss of Cry1Ac binding to membrane vesicles. ABC proteins are integral membrane proteins with many functions, including export of toxic molecules from the cell, but have not been implicated in the mode of action of Bt toxins before. The reduction in toxin binding due to the inactivating mutation suggests that ABCC2 is involved in membrane integration of the toxin pore. Our findings suggest that ABC proteins may play a key role in the mode of action of Bt toxins and that ABC protein mutations can confer high levels of resistance that could threaten the continued utilization of Bt–expressing crops. However, such mutations may impose a physiological cost on resistant insects, by reducing export of other toxins such as plant secondary compounds from the cell. This weakness could be exploited to manage this mechanism of Bt resistance in the field.

Evolutionary ecology of insect adaptation to Bt crops

Abstract: Transgenic crops producing Bacillus thuringiensis (Bt) toxins are used worldwide to control major pests of corn and cotton. Development of strategies to delay the evolution of pest resistance to Bt crops requires an understanding of factors affecting responses to natural selection, which include variation in survival on Bt crops, heritability of resistance, and fitness advantages associated with resistance mutations. The two main strategies adopted for delaying resistance are the refuge and pyramid strategies. Both can reduce heritability of resistance, but pyramids can also delay resistance by reducing genetic variation for resistance. Seasonal declines in the concentration of Bt toxins in transgenic cultivars, however, can increase the heritability of resistance. The fitness advantages associated with resistance mutations can be reduced by agronomic practices, including increasing refuge size, manipulating refuges to increase fitness costs, and manipulating Bt cultivars to reduce fitness of resistant individuals. Manipulating costs and fitness of resistant individuals on transgenic insecticidal crops may be especially important for thwarting evolution of resistance in haplodiploid and parthenogenetic pests. Field-evolved resistance to Bt crops in only five pests during the last 14 years suggests that the refuge strategy has successfully delayed resistance, but the accumulation of resistant pests could accelerate.

Suppressing resistance to Bt cotton with sterile insect releases.

Abstract: Adaptation of insect pests to tolerate Bacillus thuringiensis (Bt) insecticidal proteins threatens to reduce the efficacy of Bt crops. Evidence from an extensive four-year field trial indicates that the release of sterile pink bollworm moths suppresses the emergence of resistance to transgenic Bt cotton, while helping to eradicate the pest.

Novel Vip3A Bacillus thuringiensis (Bt) maize approaches high-dose efficacy against Helicoverpa zea (Lepidoptera: Noctuidae) under field conditions: Implications for resistance management.

Abstract: Sweet corn, Zea mays L., transformed to express a novel vegetative insecticidal protein, Vip3A (event MIR162, Syngenta Seeds, Inc..), produced by the bacterium, Bacillus thuringiensis (Bt), was evaluated over four field seasons in Maryland and two field seasons in Minnesota for efficacy against the corn earworm, Helicoverpa zea (Boddie). Hybrids expressing the Vip3A protein and pyramided in hybrids also expressing the Cry1Ab Bt protein (event Bt11, ATTRIBUTE(®), Syngenta Seeds, Inc.) were compared to hybrids expressing only Cry1Ab or to genetically similar non-Bt hybrids each year. In addition to H. zea efficacy, results for Ostrinia nubilalis (Hubner) and Spodoptera frugiperda (J.E. Smith) are presented. Over all years and locations, the non-Bt hybrids, without insecticide protection, averaged between 43 and 100% ears infested with a range of 0.24 to 1.74 H. zea larvae per ear. By comparison, in the pyramided Vip3A x Cry1Ab hybrids, no larvae were found and only minimal kernel damage (likely due to other insect pests) was recorded. Hybrids expressing only Cry1Ab incurred a moderate level of H. zea feeding damage, with surviving larvae mostly limited to the first or second instar as a result of previously documented growth inhibition from Cry1Ab. These results suggest that the Vip3A protein, pyramided with Cry1Ab, appears to provide the first "high-dose" under field conditions and will be valuable for ongoing resistance management.

Hardy Cotton-Munching Pests Are Latest Blow to GM Crops

Abstract: Monsanto has revealed that a common insect pest has developed resistance to its flagship genetically modified (GM) product in India. The agricultural biotechnology leader says it "detected unusual survival" of pink bollworms that fed on cotton containing the Cry1Ac gene from the bacterium Bacillus thuringiensis, which codes for a protein that9s toxic to many insect pests. In a statement to Science, Monsanto claims that the finding from western India "is the first case of field-relevant resistance to Cry1Ac products, anywhere in the world."

Environmental factors determining the epidemiology and population genetic structure of the Bacillus cereus group in the field.

Abstract: Bacillus thuringiensis (Bt) and its insecticidal toxins are widely exploited in microbial biopesticides and genetically modified crops. Its population biology is, however, poorly understood. Important issues for the safe, sustainable exploitation of Bt include understanding how selection maintains expression of insecticidal toxins in nature, whether entomopathogenic Bt is ecologically distinct from related human pathogens in the Bacillus cereus group, and how the use of microbial pesticides alters natural bacterial populations. We addressed these questions with a MLST scheme applied to a field experiment in which we excluded/added insect hosts and microbial pesticides in a factorial design. The presence of insects increased the density of Bt/B. cereus in the soil and the proportion of strains expressing insecticidal toxins. We found a near-epidemic population structure dominated by a single entomopathogenic genotype (ST8) in sprayed and unsprayed enclosures. Biopesticidal ST8 proliferated in hosts after spraying but was also found naturally associated with leaves more than any other genotype. In an independent experiment several ST8 isolates proved better than a range of non-pathogenic STs at endophytic and epiphytic colonization of seedlings from soil. This is the first experimental demonstration of Bt behaving as a specialized insect pathogen in the field. These data provide a basis for understanding both Bt ecology and the influence of anthropogenic factors on Bt populations. This natural population of Bt showed habitat associations and a population structure that differed markedly from previous MLST studies of less ecologically coherent B. cereus sample collections. The host-specific adaptations of ST8, its close association with its toxin plasmid and its high prevalence within its clade are analogous to the biology of Bacillus anthracis. This prevalence also suggests that selection for resistance to the insecticidal toxins of ST8 will have been stronger than for other toxin classes.

Incipient resistance of Helicoverpa punctigera to the Cry2Ab Bt toxin in Bollgard II cotton.

Abstract: Combinations of dissimilar insecticidal proteins (“pyramids”) within transgenic plants are predicted to delay the evolution of pest resistance for significantly longer than crops expressing a single transgene. Field-evolved resistance to Bacillus thuringiensis (Bt) transgenic crops has been reported for first generation, single-toxin varieties and the Cry1 class of proteins. Our five year data set shows a significant exponential increase in the frequency of alleles conferring Cry2Ab resistance in Australian field populations of Helicoverpa punctigera since the adoption of a second generation, two-toxin Bt cotton expressing this insecticidal protein. Furthermore, the frequency of cry2Ab resistance alleles in populations from cropping areas is 8-fold higher than that found for populations from non-cropping regions. This report of field evolved resistance to a protein in a dual-toxin Bt-crop has precisely fulfilled the intended function of monitoring for resistance; namely, to provide an early warning of increases in frequencies that may lead to potential failures of the transgenic technology. Furthermore, it demonstrates that pyramids are not ‘bullet proof’ and that rapid evolution to Bt toxins in the Cry2 class is possible.

Lepidopteran-Active Variable-Region Sequence Imparts Coleopteran Activity in eCry3.1Ab, an Engineered Bacillus thuringiensis Hybrid Insecticidal Protein

Abstract: A unique, coleopteran-active protein, termed eCry3.1Ab, was generated following variable-region exchange of a Bacillus thuringiensis lepidopteran-active protein, Cry1Ab, with a Cry3A region. Our results support the hypothesis that this variable-region exchange is responsible for imparting strong bioactivity against the larvae of western corn rootworm (WCR) (Diabrotica virgifera virgifera LeConte), a pest species which is not susceptible to either parent protein sequence. This study demonstrates the potential of successfully engineering a portion(s) of a lepidopteran-active B. thuringiensis sequence so that it has activity against coleopterans. Further elucidation of the eCry3.1Ab activity indicated the importance of variable regions 4 to 6 that were derived from Cry1Ab instead of Cry1Ac. There was some flexibility in making domain III of engineered hybrid insecticidal proteins even more Cry1Ab-like and retaining activity, while there was less flexibility in making domain III more Cry3A-like and retaining activity. In vitro binding studies with brush border membrane vesicles demonstrated that there was specific binding of chymotrypsin-processed modified Cry3A (mCry3A), which was not diminished by addition of a 100-fold molar excess of chymotrypsin-processed eCry3.1Ab or unprocessed eCry3.1Ab. In addition, in the converse experiment, specific binding of chymotrypsin-processed eCry3.1Ab was not diminished by the presence of a 75-fold molar excess of chymotrypsin-processed mCry3A. These data support the hypothesis that eCry3.1Ab can interact with different binding sites than the activated form of mCry3A in the WCR brush border and may provide a different mode of action from the standpoint of resistance management.

Binding Site Alteration Is Responsible for Field-Isolated Resistance to Bacillus thuringiensis Cry2A Insecticidal Proteins in Two Helicoverpa Species

Abstract: Background Evolution of resistance by target pests is the main threat to the long-term efficacy of crops expressing Bacillus thuringiensis (Bt) insecticidal proteins. Cry2 proteins play a pivotal role in current Bt spray formulations and transgenic crops and they complement Cry1A proteins because of their different mode of action. Their presence is critical in the control of those lepidopteran species, such as Helicoverpa spp., which are not highly susceptible to Cry1A proteins. In Australia, a transgenic variety of cotton expressing Cry1Ac and Cry2Ab (Bollgard II) comprises at least 80% of the total cotton area. Prior to the widespread adoption of Bollgard II, the frequency of alleles conferring resistance to Cry2Ab in field populations of Helicoverpa armigera and Helicoverpa punctigera was significantly higher than anticipated. Colonies established from survivors of F2 screens against Cry2Ab are highly resistant to this toxin, but susceptible to Cry1Ac.

Spatial and temporal variability in host use by Helicoverpa zea as measured by analyses of stable carbon isotope ratios and gossypol residues.

Abstract: Summary 1. A high dose ⁄ refuge strategy has been adopted in the USA to manage the risk of Bacillus thuringiensis (Bt) resistance in target pests such as the cotton bollworm (CBW), Helicoverpa zea (Boddie) in transgenic Bt cotton Gossypium hirsutum L. Structured refuges, consisting of non-Bt cotton, have been a mandated part of this strategy to produce non-selected insects that are temporally and spatially synchronous with insects from the Bt crop, diluting Bt resistance alleles through mating. However, the bollworm is highly polyphagous and exploits a large number of crop and weedy hosts concurrently with Bt cotton.

Susceptibility of Isofamilies of Spodoptera frugiperda (Lepidoptera: Noctuidae) to Cry1Ac and Cry1Fa Proteins of Bacillus thuringiensis

Abstract: The fall armyworm, Spodoptera frugiperda (J. E. Smith), is one of the most important insect pests on the American continent. Its control has relied primarily on multiple applications of insecticides that can amount to 1,000 g of active ingredient per hectare on some of approximately 30 crops the insect damages. The use of genetically engineered crops that express Bacillus thuringiensis (Bt) Berliner toxins, Bt-corn, Zee meys L.; and Bt-cotton, Gossypium hirsutum L.; are other ways to control this insect. However, fall armyworm is one of the Lepidoptera species least susceptible to Bt proteins, and a case of high tolerance to Bt-corn has already being reported. We found the susceptibility to Cry1Ac and Cry1Fa proteins of Bt in 133 isofamilies from five regions of three countries was similar to the susceptibility of two Bt-susceptible laboratory colonies to these proteins. Four isofamilies from Puerto Rico were very tolerant to Cry1Fa and not so tolerant to Cry1Ac. Two of the four isofamilies were backcrossed with a Bt-susceptible laboratory colony and their progeny was as susceptible to both Bt proteins as was the Bt-susceptible colony, indicating that resistance to Bt is a recessive trait.

A mathematical model of exposure of non- target Lepidoptera to Bt-maize pollen expressing Cry1Ab within Europe

Abstract: Genetically modified (GM) maize MON810 expresses a Cry1Ab insecticidal protein, derived from Bacillus thuringiensis (Bt), toxic to lepidopteran target pests such as Ostrinia nubilalis. An environmental risk to non-target Lepidoptera from this GM crop is exposure to harmful amounts of Bt-containing pollen deposited on host plants in or near MON810 fields. An 11-parameter mathematical model analysed exposure of larvae of three non-target species: the butterflies Inachis io (L.), Vanessa atalanta (L.) and moth Plutella xylostella (L.), in 11 representative maize cultivation regions in four European countries. A mortality-dose relationship was integrated with a dose-distance relationship to estimate mortality both within the maize MON810 crop and within the field margin at varying distances from the crop edge. Mortality estimates were adjusted to allow for physical effects; the lack of temporal coincidence between the susceptible larval stage concerned and the period over which maize MON810 pollen is shed; and seven further parameters concerned with maize agronomy and host-plant ecology. Sublethal effects were estimated and allowance made for aggregated pollen deposition. Estimated environmental impact was low: in all regions, the calculated mortality rate for worst-case scenarios was less than one individual in every 1572 for the butterflies and one in 392 for the moth.

Complete Genome Sequence of Bacillus thuringiensis Mutant Strain BMB171

Abstract: Bacillus thuringiensis has been widely used as a biopesticide for a long time. Here we report the finished and annotated genome sequence of B. thuringiensis mutant strain BMB171, an acrystalliferous mutant strain with a high transformation frequency obtained and stocked in our laboratory.

GENOME ANNOUNCEMENTS Complete Genome Sequence of Bacillus thuringiensis Mutant Strain BMB171

Abstract: Bacillus thuringiensis is an insect pathogen which is widely used as a biopesticide due to its various endogenous crystal proteins and spores (12). To improve the virulence and practical effectiveness of B. thuringiensis, genetic transformation of different genes with beneficial traits is a fundamental procedure. Simultaneously, genetic transformation can facilitate functional genomic research. However, wild-type strains are not suitable to be used as recipient strains because of low transformation efficiency. This obstacle is mainly caused by the thick cell wall layer of B. thuringiensis together with multiple plasmids inside the cell, which harbor genes encoding insecticidal crystal proteins. We used the method of elevating the growth temperature and adding 0.05% sodium dodecyl sulfate to treat several parental strains and finally obtained mutant strain BMB171, with no resident plasmid, from wild-type crystalliferous strain YBT1463 (9). The electrotransformation frequency of mutant

Constitutive activation of the midgut response to Bacillus thuringiensis in Bt-resistant Spodoptera exigua.

Abstract: Bacillus thuringiensis is the most effective microbial control agent for controlling numerous species from different insect orders. The main threat for the long term use of B. thuringiensis in pest control is the ability of insects to develop resistance. Thus, the identification of insect genes involved in conferring resistance is of paramount importance. A colony of Spodoptera exigua (Lepidoptera: Noctuidae) was selected for 15 years in the laboratory for resistance to Xentari™, a B. thuringiensis-based insecticide, reaching a final resistance level of greater than 1,000-fold. Around 600 midgut ESTs were analyzed by DNA-macroarray in order to find differences in midgut gene expression between susceptible and resistant insects. Among the differentially expressed genes, repat and arylphorin were identified and their increased expression was correlated with B. thuringiensis resistance. We also found overlap among genes that were constitutively over-expressed in resistant insects with genes that were up-regulated in susceptible insects after exposure to Xentari™, suggesting a permanent activation of the response to Xentari™ in resistant insects. Increased aminopeptidase activity in the lumen of resistant insects in the absence of exposure to Xentari™ corroborated the hypothesis of permanent activation of response genes. Increase in midgut proliferation has been proposed as a mechanism of response to pathogens in the adult from several insect species. Analysis of S. exigua larvae revealed that midgut proliferation was neither increased in resistant insects nor induced by exposure of susceptible larvae to Xentari™, suggesting that mechanisms other than midgut proliferation are involved in the response to B. thuringiensis by S. exigua larvae.

Exposure and nontarget effects of transgenic Bt corn debris in streams.

Abstract: Corn (Zea mays L.) transformed with a gene from the bacterium Bacillus thuringiensis (Bt) comprises 49% of all corn in the United States. The input of senesced corn tissue expressing the Bt gene may impact stream-inhabiting invertebrates that process plant debris, especially trichopteran species related to the target group of lepidopteran pests. Our goal was to assess risk associated with transgenic corn debris entering streams. First, we show the input of corn tissue after harvest was extended over months in a stream. Second, using laboratory bioassays based on European corn borer [Ostrinia nubilalis (Hubner)], we found no bioactivity of Cry1Ab protein in senesced corn tissue after 2 wk of exposure to terrestrial or aquatic environments. Third, we show that Bt near-isolines modify growth and survivorship of some species of invertebrates. Of the four nontarget invertebrate species fed Bt near-isolines, growth of two closely related trichopterans was not negatively affected, whereas a tipulid crane fly exhibited reduced growth rates, and an isopod exhibited reduced growth and survivorship on the Cry1Ab near-isoline but not on the stacked Cry1Ab + Cry3Bb1 near-isoline. Because of lack of evidence of bioactivity of Bt after 2 wk and because of lack of nontarget effects on the stacked near-isoline, we suggest that tissue-mediated differences, and not the presence of the Cry1Ab protein, caused the different responses among the species. Overall, our results provide evidence that adverse effects to aquatic nontarget shredders involve complex interactions arising from plant genetics and environment that cannot be ascribed to the presence of Cry1Ab proteins.

A synthesis of laboratory and field studies on the effects of transgenic Bacillus thuringiensis (Bt) maize on non-target Lepidoptera

Abstract: One of the major applications of transgenic crops in agriculture are the so-called Bacillus thuringiensis Berliner (Bt) plants, in particular Bt maizes, which produce insecticidal Cry proteins that target specific orders, such as the Lepidoptera or Coleoptera. We reviewed publications that reported on the direct toxic effects of Bt-maize and/or Cry proteins of current Bt-maize events on larvae of non-target butterflies and moths (Lepidoptera). In total, 20 peer-reviewed publications were identified, of which 16 papers contributed laboratory-based data and seven field-based data. An adverse effect on caterpillars was recorded in 52% of all laboratory-based and in 21% of all field-based observations. The variables most often studied and having the highest occurrence of effects were larval survival, body mass, and developmental time. Parameters of the adult stage were under-represented in the studies. Overall, 11 lepidopteran species were tested. The majority of the studies originated from the USA, with the Monarch butterfly being the most studied, whereas other species and other parts of the world were widely neglected. Laboratory experiments were often run under unrealistic conditions from an ecological point of view. Although the papers we reviewed indicated a potential hazard for Lepidoptera that are exposed to and feed on lepidopteran-specific Bt-maize pollen, a general conclusion on the level of risk for butterflies and moths cannot as yet be drawn. A comprehensive risk characterization would require thorough hazard identification, exposure assessment, and impact assessment. However, our review showed that even the basic level of hazard characterization is as yet incomplete. Reasons for this are the still-limited numbers of publications and concurrent lack of knowledge, the restriction of data to only a few species, the over-representation of North American species, and the identified limitations of both laboratory and field experiments. The findings of this review suggest that more realistic, ecologically meaningful, and detailed experiments and analyses are crucial to improve the present assessment of Bt-maize cultivation effects on Lepidoptera.

An α‐amylase is a novel receptor for Bacillus thuringiensis ssp. israelensis Cry4Ba and Cry11Aa toxins in the malaria vector mosquito Anopheles albimanus (Diptera: Culicidae)

Abstract: Bacillus thuringiensis ssp. israelensis (Bti) produces four Cry toxins (Cry4Aa, Cry4Ba, Cry10Aa and Cry11Aa), and two Cyt proteins (Cyt1Aa and Cyt2Ba), toxic to mosquito-larvae of the genus Aedes, Anopheles and Culex, important human disease vectors that transmit dengue virus, malaria and filarial parasites respectively. Previous work showed that Bti is highly toxic to Anopheles albimanus, the main vector for transmission of malaria in Mexico. In this work, we analysed the toxicity of isolated Cry proteins of Bti and identified an An. albimanus midgut protein as a putative Cry4Ba and Cry11Aa receptor molecule. Biossays showed that Cry4Ba and Cry11Aa of Bti are toxic to An. albimanus larvae. Ligand blot assays indicated that a 70 kDa glycosylphosphatidylinositol-anchored protein present in midgut brush border membrane vesicles of An. albimanus interacts with Cry4Ba and Cry11Aa toxins. This protein was identified as an alpha-amylase by mass spectrometry and enzymatic activity assays. The cDNA that codes for the alpha-amylase was cloned by means of 5'- and 3'-RACE experiments. Recombinant alpha-amylase expressed in Escherichia coli specifically binds Cry4Ba and Cry11Aa toxins.

RNAi-mediated knockdown of a Spodoptera frugiperda trypsin-like serine-protease gene reduces susceptibility to a Bacillus thuringiensis Cry1Ca1 protoxin

Abstract: SfT6 has been identified in a subtracted cDNA library of Spodoptera frugiperda larval midgut transcripts as a serine-protease gene downregulated within 24 h of intoxication with Bacillus thuringiensis Cry1Ca1 protein In the present study, the specific role of SfT6 during Cry1Ca1 intoxication was investigated by RT-PCR and in vivo RNA interference Quantitative real-time RT-PCR analysis showed SfT6 mRNA levels in the midgut tissue were significantly reduced after injecting or feeding 4th-instar larvae with specific long-size dsRNA Gut juice-mediated in vitro protoxin activation and susceptibility for Cry1Ca1 were investigated in Sft6-knockdown larvae and compared with control treated with nonspecific dsRNA Our results demonstrate SfT6 plays a determinant role in Cry1Ca1 toxicity against S frugiperda since a decreased expression caused a reduced protoxin activation by larval gut juice and reduced susceptibility of insects to toxin in bioassays We propose SfT6 downregulation occurring at the early stages of Cry1Ca1 intoxication is part of a complex and multifaceted defensive mechanism triggered in the insect gut to withstand B thuringiensis pathogenesis

Field-evolved resistance to Bt cotton: bollworm in the U.S. and pink bollworm in India.

Abstract: . Some field populations of bollworm, Helicoverpa zea (Boddie), in the southeastern U.S. have evolved resistance to Bacillus thuringiensis Berliner (Bt) toxins Cry1Ac and Cry2Ab produced by transgenic cotton, Gossypium hirsutum L. Decreased susceptibility to Cry1Ac, the toxin in first-generation Bt cotton (Bollgard), was detected in laboratory diet bioassays and was associated with increased survival on Bollgard plant tissues and control problems in the field. Bollgard cotton is no longer registered in the U.S. and has been replaced primarily by Bt cotton that produces Cry1Ac and Cry2Ab (Bollgard II). Laboratory diet bioassay results showed that the percentage of field populations resistant to Cry2Ab increased from 0% in 2002 to 50% in 2005. In bioassays from 2003 to 2006, 14 field-derived strains of bollworm had <50% mortality at 150 µg Cry2Ab per milliliter of diet, the highest toxin concentration tested. As with the recently reported resistance of pink bollworm, Pectinophora gossypiella (Saunde...

Decomposition dynamics and structural plant components of genetically modified Bt maize leaves do not differ from leaves of conventional hybrids

Abstract: The cultivation of genetically modified Bt maize has raised environmental concerns, as large amounts of plant residues remain in the field and may negatively impact the soil ecosystem. In a field experiment, decomposition of leaf residues from three genetically modified (two expressing the Cry1Ab, one the Cry3Bb1 protein) and six non-transgenic hybrids (the three corresponding non-transformed near-isolines and three conventional hybrids) was investigated using litterbags. To elucidate the mechanisms that cause differences in plant decomposition, structural plant components (i.e., C:N ratio, lignin, cellulose, hemicellulose) were examined. Furthermore, Cry1Ab and Cry3Bb1 protein concentrations in maize leaf residues were measured from harvest to the next growing season. While leaf residue decomposition in transgenic and non-transgenic plants was similar, differences among conventional cultivars were evident. Similarly, plant components among conventional hybrids differed more than between transgenic and non-transgenic hybrids. Moreover, differences in senescent plant material collected directly from plants were larger than after exposure to soil for 5 months. While the concentration of Cry3Bb1 was higher in senescent maize leaves than that of Cry1Ab, degradation was faster, indicating that Cry3Bb1 has a shorter persistence in plant residues. As decomposition patterns of Bt-transgenic maize were shown to be well within the range of common conventional hybrids, there is no indication of ecologically relevant, adverse effects on the activity of the decomposer community.

Pollen- and seed-mediated transgene flow in commercial cotton seed production fields.

Abstract: Background Characterizing the spatial patterns of gene flow from transgenic crops is challenging, making it difficult to design containment strategies for markets that regulate the adventitious presence of transgenes. Insecticidal Bacillus thuringiensis (Bt) cotton is planted on millions of hectares annually and is a potential source of transgene flow.

Interaction of Bacillus thuringiensis Vegetative Insecticidal Protein with Ribosomal S2 Protein Triggers Larvicidal Activity in Spodoptera frugiperda

Abstract: Vegetative insecticidal protein (Vip3A) is synthesized as an extracellular insecticidal toxin by certain strains of Bacillus thuringiensis. Vip3A is active against several lepidopteran pests of crops. Polyphagous pest, Spodoptera frugiperda, and its cell line Sf21 are sensitive for lyses to Vip3A. Screening of cDNA library prepared from Sf21 cells through yeast two-hybrid system with Vip3A as bait identified ribosomal protein S2 as a toxicity-mediating interacting partner protein. The Vip3A-ribosomal-S2 protein interaction was validated by in vitro pulldown assays and by RNA interference-induced knockdown experiments. Knockdown of expression of S2 protein in Sf21 cells resulted in reduced toxicity of the Vip3A protein. These observations were further extended to adult fifth-instar larvae of Spodoptera litura. Knockdown of S2 expression by injecting corresponding double-stranded RNA resulted in reduced mortality of larvae to Vip3A toxin. Intracellular visualization of S2 protein and Vip3A through confocal microscopy revealed their interaction and localization in cytoplasm and surface of Sf21 cells.

Risk assessment of toxins derived from Bacillus thuringiensis—synergism, efficacy, and selectivity

Abstract: Background, aim, and scope This review deals with publications concerning the mode of action of Bt proteins and their potential synergism with extrinsic factors. The aim was to assess the impact of those factors especially regarding selectivity and efficacy of Bt toxins and to discuss possible gaps in current risk assessment of genetically engineered plants expressing Bt toxins.

Cross‐resistance of Cry1Ab‐selected Asian corn borer to other Cry toxins

Abstract: The Asian corn borer, Ostrinia furnacalis (Guene´e) (Lepidoptera: Crambidae), is the most important insect pest of maize in China. Despite the proven track record of biotech Bacillus thuringiensis (Bt) maize to provide an effective means to control lepidopteran pests such as the European corn borer Ostrinia nubilalis (Hu¨ bner), these crops have not been commercialized in China. However, Cry1Ab- and Cry1F expressing maize derived from genetic transformation events MON810 (Monsanto), Bt11 (Syngenta) and TC1507 (Pioneer, DuPont) as well as Cry1Ah-, Cry1A- (a Cry1Ab/Cry1Ac hybrid toxin), and Cry1Ac-, Cry1Ie-expressing maize developed by the Chinese institutions have been shown to effectively suppress O. furnacalis in field trials. The development of resistance in target pests is a major threat to the use of Bt maize. A strain of O. furnacalis selected with Cry1Ab protein incorporated into artificial diet developed more than 100-fold resistance to Cry1Ab after 35 generations of selection, and readily consumed Cry1Ab-expressing maize silks. Although LC50s of the selected insect strain were not progressively increased, and even decreased in the following generations, these insects (ACB-AbR) could survive on Cry1Ab-expressing maize tissue after 51 generations of selection. In addition, susceptibility of ACB-AbR to a number of other Bt toxins to which the selected strains had not previously been exposed, was significantly decreased. The highest level of cross-resistance was observed with Cry1Ah (131-fold), followed by Cry1Ac (36-fold). A low level of cross-resistance (6-fold) to Cry1F was also detected. In contrast, ACB-AbR was equally susceptible to Cry1Ie as the unselected control strain. These results indicate that the availability of multiple toxins could improve resistance management strategies, provided that the potential for cross-resistance among toxins is not an issue.