Showing papers on "Bacillus thuringiensis published in 2005"
TL;DR: This review provides information on the genus Bacillus and a variety of species, but the primary focus is on the B. cereus strains and toxins that are involved in foodborne illness.
460 citations
TL;DR: Risk assessment analyses show that the natural refuges derived from the mixed-planting system of cotton, corn, soybean, and peanut on small-scale, single-family-owned farms play an important function in delaying evolution of cotton bollworm resistance, and that no trend toward Bt Cotton resistance has been apparent despite intensive planting of Bt cotton over the past several years.
Abstract: The development of cotton pest management practices in China has followed a pattern seen for many crops that rely heavily on insecticides. Helicoverpa armigera resistance to chemical pesticides resulted in the unprecedented pest densities of the early 1990s. Transgenic cotton that expresses a gene derived from the bacterium Bacillus thuringiensis (Bt) has been deployed for combating H. armigera since 1997. The pest management tactics associated with Bt cotton have resulted in a drastic reduction in insecticide use, which usually results in a significant increase in populations of beneficial insects and thus contributes to the improvement of the natural control of some pests. Risk assessment analyses show that the natural refuges derived from the mixed-planting system of cotton, corn, soybean, and peanut on small-scale, single-family-owned farms play an important function in delaying evolution of cotton bollworm resistance, and that no trend toward Bt cotton resistance has been apparent despite intensive planting of Bt cotton over the past several years.
422 citations
TL;DR: It is demonstrated that the major mechanism for Bt toxin resistance in Caenorhabditis elegans entails a loss of glycolipid carbohydrates, and evidence that insect Glycolipids are also receptors for BT toxin is presented, showing carbohydrate-dependent and relevant for toxin action in vivo.
Abstract: The development of pest resistance threatens the effectiveness of Bacillus thuringiensis (Bt) toxins used in transgenic and organic farming. Here, we demonstrate that (i) the major mechanism for Bt toxin resistance in Caenorhabditis elegans entails a loss of glycolipid carbohydrates; (ii) Bt toxin directly and specifically binds glycolipids; and (iii) this binding is carbohydrate-dependent and relevant for toxin action in vivo. These carbohydrates contain the arthroseries core conserved in insects and nematodes but lacking in vertebrates. We present evidence that insect glycolipids are also receptors for Bt toxin.
347 citations
TL;DR: The results confirmed that the cadherin gene is a preferred target for developing DNA-based monitoring of B. thuringiensis resistance in field populations of lepidopteran pests.
Abstract: A laboratory strain (GY) of Helicoverpa armigera (Hubner) was established from surviving larvae collected from transgenic cotton expressing a Bacillus thuringiensis var. kurstaki insecticidal protein (Bt cotton) in Gaoyang County, Hebei Province, People's Republic of China, in 2001. The GYBT strain was derived from the GY strain through 28 generations of selection with activated Cry1Ac delivered by diet surface contamination. When resistance to Cry1Ac in the GYBT strain increased to 564-fold after selection, we detected high levels of cross-resistance to Cry1Aa (103-fold) and Cry1Ab (>46-fold) in the GYBT strain with reference to those in the GY strain. The GYBT strain had a low level of cross-resistance to B. thuringiensis var. kurstaki formulation (Btk) (5-fold) and no cross-resistance to Cry2Aa (1.4-fold). Genetic analysis showed that Cry1Ac resistance in the GYBT strain was controlled by one autosomal and incompletely recessive gene. The cross-resistance pattern and inheritance mode suggest that the Cry1Ac resistance in the GYBT strain of H. armigera belongs to "mode 1," the most common type of lepidopteran resistance to B. thuringiensis toxins. A cadherin gene was cloned and sequenced from both the GY and GYBT strains. Disruption of the cadherin gene by a premature stop codon was associated with a high level of Cry1Ac resistance in H. armigera. Tight linkage between Cry1Ac resistance and the cadherin locus was observed in a backcross analysis. Together with previous evidence found with Heliothis virescens and Pectinophora gossypiella, our results confirmed that the cadherin gene is a preferred target for developing DNA-based monitoring of B. thuringiensis resistance in field populations of lepidopteran pests.
307 citations
TL;DR: GM plants expressing other insect-resistant proteins that have a broader spectrum of activity have been tested on only a limited number of nontarget species and appear to have little impact on soil biota such as earthworms, collembolans, and general soil microflora.
Abstract: Insect resistance, based on Bacillus thuringiensis (Bt) endotoxins, is the second most widely used trait (after herbicide resistance) in commercial genetically modified (GM) crops. Other modifications for insect resistance, such as proteinase inhibitors and lectins, are also being used in many experimental crops. The extensive testing on nontarget plant-feeding insects and beneficial species that has accompanied the long-term and wide-scale use of Bt plants has not detected significant adverse effects. GM plants expressing other insect-resistant proteins that have a broader spectrum of activity have been tested on only a limited number of nontarget species. Little is known about the persistence of transgene-derived proteins in soil, with the exception of Bt endotoxins, which can persist in soil for several months. Bt plants appear to have little impact on soil biota such as earthworms, collembolans, and general soil microflora. Further research is required on the effects of GM plants on soil processes such as decomposition. Assessment of nontarget impacts is an essential part of the risk assessment process for insect-resistant GM plants.
302 citations
TL;DR: Transgenic maize (Zea mays L.) hybrids that control CRW are described, created with a Cry3Bb1 Bacillus thuringiensis (Bt) variant that is approximately eight times more lethal to corn rootworm larvae than the wild-type protein.
Abstract: The corn rootworm (CRW; Diabrotica spp.) is one of the most serious pests of corn in the USA. Chemical insecticides and crop rotation have been the only two options available to growers for managing CRW. Unfortunately, both of these tactics can be ineffective as a result of either resistance or behavioral modifications. In this paper, we describe transgenic maize (Zea mays L.) hybrids that control CRW. These hybrids were created with a Cry3Bb1 Bacillus thuringiensis (Bt) variant that is approximately eight times more lethal to corn rootworm larvae than the wild-type protein. A DNA vector containing the modified cry3Bb1 gene was placed under control of a root-enhanced promoter (4-AS1) and was introduced into embryonic maize cells by microprojectile bombardment. Described here is the molecular genetic characterization, protein expression levels, and field performance of the recently commercialized MON863 hybrids.
252 citations
TL;DR: Evidence is provided that Cyt1Aa functions as a receptor of Cry11Aa, a highly effective pathogenic bacterium that produces a toxin and also its functional receptor, promoting toxin binding to the target membrane and causing toxicity.
Abstract: Bacillus thuringiensis subsp. israelensis produces crystal proteins, Cry (4Aa, 4Ba, 10Aa, and 11Aa) and Cyt (1Aa and 2Ba) proteins, toxic to mosquito vectors of human diseases. Cyt1Aa overcomes insect resistance to Cry11Aa and Cry4 toxins and synergizes the toxicity of these toxins. However, the molecular mechanism of synergism remains unsolved. Here, we provide evidence that Cyt1Aa functions as a receptor of Cry11Aa. Sequential-binding analysis of Cyt1Aa and Cry11Aa revealed that Cyt1Aa binding to Aedes aegypti brush border membrane vesicles enhanced the binding of biotinylated-Cry11Aa. The Cyt1Aa- and Cry11Aa-binding epitopes were mapped by means of the yeast two-hybrid system, peptide arrays, and heterologous competition assays with synthetic peptides. Two exposed regions in Cyt1Aa, loop β6-αE and part of β7, bind Cry11Aa. On the other side, Cry11Aa binds Cyt1Aa proteins by means of domain II-loop α8 and β-4, which are also involved in midgut receptor interaction. Characterization of single-point mutations in Cry11Aa and Cyt1Aa revealed key Cry11Aa (S259 and E266) and Cyt1Aa (K198, E204 and K225) residues involved in the interaction of both proteins and in synergism. Additionally, a Cyt1Aa loop β6-αE mutant (K198A) with enhanced synergism to Cry11Aa was isolated. Data provided here strongly indicates that Cyt1Aa synergizes or suppresses resistance to Cry11Aa toxin by functioning as a membrane-bound receptor. Bacillus thuringiensis subsp. israelensis is a highly effective pathogenic bacterium because it produces a toxin and also its functional receptor, promoting toxin binding to the target membrane and causing toxicity.
217 citations
TL;DR: The multimodal approach utilized in this work may be applied in long-term field studies aimed at monitoring the real hazard of genetically modified crops and their residues on nontarget soil microbial communities.
Abstract: A polyphasic approach has been developed to gain knowledge of suitable key indicators for the evaluation of environmental impact of genetically modified Bt 11 and Bt 176 corn lines on soil ecosystems. We assessed the effects of Bt corn (which constitutively expresses the insecticidal toxin from Bacillus thuringiensis, encoded by the truncated Cry1Ab gene) and non-Bt corn plants and their residues on rhizospheric and bulk soil eubacterial communities by means of denaturing gradient gel electrophoresis analyses of 16S rRNA genes, on the nontarget mycorrhizal symbiont Glomus mosseae, and on soil respiration. Microcosm experiments showed differences in rhizospheric eubacterial communities associated with the three corn lines and a significantly lower level of mycorrhizal colonization in Bt 176 corn roots. In greenhouse experiments, differences between Bt and non-Bt corn plants were detected in rhizospheric eubacterial communities (both total and active), in culturable rhizospheric heterotrophic bacteria, and in mycorrhizal colonization. Plant residues of transgenic plants, plowed under at harvest and kept mixed with soil for up to 4 months, affected soil respiration, bacterial communities, and mycorrhizal establishment by indigenous endophytes. The multimodal approach utilized in our work may be applied in long-term field studies aimed at monitoring the real hazard of genetically modified crops and their residues on nontarget soil microbial communities.
213 citations
TL;DR: Concurrent use of transgenic plants expressing a single and two Bt genes will select for resistance to two-gene plants more rapidly than the use of two-Gene plants alone, which has important implications for the regulation and deployment of pyramided Bt plants.
Abstract: Transgenic plants expressing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) were grown on over 13 million ha in the United States and 22.4 million ha worldwide in 2004. Preventing or slowing the evolution of resistance by insects (“resistance management”) is critical for the sustainable use of Bt crops. Plants containing two dissimilar Bt toxin genes in the same plant (“pyramided”) have the potential to delay insect resistance. However, the advantage of pyramided Bt plants for resistance management may be compromised if they share similar toxins with single-gene plants that are deployed simultaneously. We tested this hypothesis using a unique model system composed of broccoli plants transformed to express different Cry toxins (Cry1Ac, Cry1C, or both) and a synthetic population of the diamondback moth (Plutella xylostella) carrying genes for resistance to Cry1Ac and Cry1C at frequencies of ≈0.10 and 0.34, respectively. After 24–26 generations of selection in the greenhouse, the concurrent use of one- and two-gene plants resulted in control failure of both types of Bt plants. When only two-gene plants were used in the selection, no or few insects survived on one- or two-gene Bt plants, indicating that concurrent use of transgenic plants expressing a single and two Bt genes will select for resistance to two-gene plants more rapidly than the use of two-gene plants alone. The results of this experiment agree with the predictions of a Mendelian deterministic simulation model and have important implications for the regulation and deployment of pyramided Bt plants.
209 citations
TL;DR: A synthesis of experimental and modeling results suggests that this delay in resistance to pink bollworm can be explained by refuges of cotton without Bt toxin, recessive inheritance of resistance, incomplete resistance, and fitness costs associated with resistance.
Abstract: Transgenic crops producing Bacillus thuringiensis (Bt) toxins kill some key insect pests and thus can reduce reliance on insecticides. Widespread planting of such Bt crops increased concerns that their usefulness would be cut short by rapid evolution of resistance to Bt toxins by pests. Pink bollworm (Pectinophora gossypiella) is a major pest that has experienced intense selection for resistance to Bt cotton in Arizona since 1997. We monitored pink bollworm resistance to Bt toxin for 8 years with laboratory bioassays of strains derived annually from 10-17 cotton fields statewide. Bioassay results show no net increase from 1997 to 2004 in the mean frequency of pink bollworm resistance to Bt toxin. A synthesis of experimental and modeling results suggests that this delay in resistance can be explained by refuges of cotton without Bt toxin, recessive inheritance of resistance, incomplete resistance, and fitness costs associated with resistance.
206 citations
TL;DR: It is demonstrated here that a population of Australian H. armigera has developed resistance to Cry1Ac toxin (275-fold); in vitro studies employing surface plasmon resonance technology and other biochemical techniques demonstrated that resistant strain esterase could bind toCry1Ac protoxin and activated toxin.
Abstract: In Australia, the cotton bollworm, Helicoverpa armigera, has a long history of resistance to conventional insecticides. Transgenic cotton (expressing the Bacillus thuringiensis toxin Cry1Ac) has been grown for H. armigera control since 1996. It is demonstrated here that a population of Australian H. armigera has developed resistance to Cry1Ac toxin (275-fold). Some 70% of resistant H. armigera larvae were able to survive on Cry1Ac transgenic cotton (Ingard) The resistance phenotype is inherited as an autosomal semidominant trait. Resistance was associated with elevated esterase levels, which cosegregated with resistance. In vitro studies employing surface plasmon resonance technology and other biochemical techniques demonstrated that resistant strain esterase could bind to Cry1Ac protoxin and activated toxin. In vivo studies showed that Cry1Ac-resistant larvae fed Cy1Ac transgenic cotton or Cry1Ac-treated artificial diet had lower esterase activity than non-Cry1Ac-fed larvae. A resistance mechanism in which esterase sequesters Cry1Ac is proposed.
TL;DR: The degradation of the biomass of all Bt plants in the absence of soil but inoculated with a microbial suspension from the same soil was significantly less than that of their respective inoculated non-Bt plants, suggesting that differences in the soil microbiota were not responsible for the differential decomposition of BtC and BtK biomass.
Abstract: Bt plants are plants that have been genetically modified to express the insecticidal proteins (e.g. Cry1Ab, Cry1Ac, Cry3A) from subspecies of the bacterium, Bacillus thuringiensis (Bt), to kill lepidopteran pests that feed on corn, rice, tobacco, canola, and cotton and coleopteran pests that feed on potato. The biomass of these transgenic Bt plants (BtC) was decomposed less in soil than the biomass of their near-isogenic non-Bt plant counterparts (BtK). Soil was amended with 0.5, 1, or 2% (wt wt K1 ) ground, dried (50 8C) leaves or stems of Bt corn plants; with 0.5% (wt wt K1 ) ground, dried biomass of Bt rice, tobacco, canola, cotton, and potato plants; with biomass of the near-isogenic plants without the respective cry genes; or not amended. The gross metabolic activity of the soil was determined by CO2 evolution. The amounts of C evolved as CO2 were significantly lower from soil microcosms amended with biomass of Bt plants than of non-Bt plants. This difference occurred with stems and leaves from two hybrids of Bt corn, one of which had a higher C:N ratio than its near-isogenic non-Bt counterpart and the other which had essentially the same C:N ratio, even when glucose, nitrogen (NH4NO3), or glucose plus nitrogen were added with the biomass. The C:N ratios of the other Bt plants (including two other hybrids of Bt corn) and their near-isogenic non-Bt counterparts were also not related to their relative biodegradation. Bt corn had a significantly higher lignin content than near-isogenic non-Bt corn. However, the lignin content of the other Bt plants, which was significantly lower than that of both Bt and non-Bt corn, was generally not statistically significantly different, although 10‐66% higher, from that of their respective non-Bt near-isolines. The numbers of culturable bacteria and fungi and the activity of representative enzymes involved in the degradation of plant biomass were not significantly different between soil amended with biomass of Bt or non-Bt corn. The degradation of the biomass of all Bt plants in the absence of soil but inoculated with a microbial suspension from the same soil was also significantly less than that of their respective inoculated non-Bt plants. The addition of streptomycin, cycloheximide, or both to the soil suspension did not alter the relative degradation of BtC and BtK biomass, suggesting that differences in the soil microbiota were not responsible for the differential decomposition of BtC and BtK biomass. All samples of soil amended with biomass of Bt plants were immunologically positive for the respective Cry proteins and toxic to the larvae of the tobacco hornworm (Manduca sexta), which was used as a representative lepidopteran in insect bioassays (no insecticidal assay was done for the Cry3A protein from potato). The ecological and environmental relevance of these findings is not clear. q 2004 Elsevier Ltd. All rights reserved.
TL;DR: Insecticidal proteins produced by various subspecies of Bacillus thuringiensis (Bt) bound rapidly and tightly onClays, both pure mined clay minerals and soil clays, on humic acids extracted from soil, and on complexes of clay and humic acid.
Abstract: Insecticidal proteins produced by various subspecies (kurstaki, tenebrionis ,a ndisraelensis )o fBacillus thuringiensis (Bt) bound rapidly and tightly on clays, both pure mined clay minerals and soil clays, on humic acids extracted from soil, and on complexes of clay and humic acids. Binding reduced susceptibility of the proteins to microbial degradation. However, bound proteins retained biological activity. Purified Cry1Ab protein and protein released from biomass of transgenic Bt corn and in root exudates of growing Bt corn (13 hybrids representing three transformation events) exhibited binding and persistence in soil. Insecticidal protein was also released in root exudates of Bt potato (Cry3A protein) and rice (Cry1Ab protein) but not in root exudates of Bt canola, cotton, and tobacco (Cry1Ac protein). Vertical movement of CrylAb protein, either purified or in root exudates or biomass of Bt corn, decreased as the concentration of the clay minerals, kaolinite or montmorillonite, in soil increased. Biomass of transgenicBt corn decomposed less in soil than biomass of near-isogenic non-Bt corn, possibly because biomass of Bt corn had a significantly higher content of lignin than biomass of non-Bt corn. Biomass of Bt canola, cotton, potato, rice, and tobacco also decomposed less than biomass of the respective near-isogenic non-Bt plants. However, the lignin content of theseBt plants, which was significantly less than that ofBt corn, was not significantly different from that of their near-isogenic non-Bt counterparts, although it was consistently higher. The Cry1Ab protein had no consistent effects on organisms (earthworms, nematodes, protozoa, bacteria, fungi) in soil or in vitro. The Cry1Ab protein was not taken up from soil by non-Bt corn, carrot, radish, or turnip grown in soil in which Bt corn had been grown or into which biomass of Bt corn had been incorporated.
TL;DR: Assessment of the nontarget effects of Bt protein toxins indicates that there is a low level of hazard to most groups of nontarget organisms, although some investigations are of limited ecological relevance.
Abstract: This paper reviews the scientific literature addressing the environmental fate and nontarget effects of the Cry protein toxins from Bacillus thuringiensis (Bt), specifically resulting from their expression in transgenic crops. Published literature on analytical methodologies for the detection and quantification of the Cry proteins in environmental matrices is also reviewed, with discussion of the adequacy of the techniques for determining the persistence and mobility of the Bt proteins. In general, assessment of the nontarget effects of Bt protein toxins indicates that there is a low level of hazard to most groups of nontarget organisms, although some investigations are of limited ecological relevance. Some published reports on the persistence of the proteins in soil show short half-lives, whereas others show low-level residues lasting for many months. Improvements in analytical methods will allow a more complete understanding of the fate and significance of Bt proteins in the environment.
TL;DR: The cloning and expression analysis of four APN cDNAs from Spodoptera exigua are reported, suggesting that the lack of APN1 expression plays a role in the resistance to Cry1Ca in this S.Exigua colony.
Abstract: Insecticidal toxins from Bacillus thuringiensis bind to receptors on midgut epithelial cells of susceptible insect larvae. Aminopeptidases N (APNs) from several insect species have been shown to be putative receptors for these toxins. Here we report the cloning and expression analysis of four APN cDNAs from Spodoptera exigua. Suppression Subtractive Hybridization (SSH) was used to construct cDNA libraries of genes that are up-and down-regulated in the midgut of last instar larvae of beet armyworm, S. exigua exposed to B. thuringiensis Cry1Ca toxin. Among the clones from the SSH libraries, cDNA fragments coding for two different APNs were obtained (APN2 and APN4). A similar procedure was employed to compare mRNA differences between susceptible and Cry1Ca resistant S. exigua. Among the clones from this last comparison, cDNA fragments belonging to a third APN (APN1) were detected. Using sequences obtained from the three APN cDNA fragments and degenerate primers for a fourth APN (APN3), the full length sequences of four S. exigua APN cDNAs were obtained. Northern blot analysis of expression of the four APNs showed complete absence of APN1 expression in the resistant insects, while the other three APNs showed similar expression levels in the resistant and susceptible insects. We have cloned and characterized four different midgut APN cDNAs from S. exigua. Expression analysis revealed the lack of expression of one of these APNs in the larvae of a Cry1Ca-resistant colony. Combined with previous evidence that shows the importance of APN in the mode of action of B. thuringiensis toxins, these results suggest that the lack of APN1 expression plays a role in the resistance to Cry1Ca in this S. exigua colony.
TL;DR: The use of model invertebrates, such as Caenorhabditis elegans and Drosophila melanogaster, as well as advances in insect genomics, are likely to accelerate efforts to clone Cry toxin resistance genes and come to a detailed and broad understanding ofCry toxin resistance.
Abstract: The Cry family of Bacillus thuringiensis insecticidal and nematicidal proteins constitutes a valuable source of environmentally benign compounds for the control of insect pests and disease agents. An understanding of Cry toxin resistance at a molecular level will be critical to the long-term utility of this technology; it may also shed light on basic mechanisms used by other bacterial toxins that target specific organisms or cell types. Selection and cross-resistance studies have confirmed that genetic adaptation can elicit varying patterns of Cry toxin resistance, which has been associated with deficient protoxin activation by host proteases, and defective Cry toxin-binding cell surface molecules, such as cadherins, aminopeptidases and glycolipids. Recent work also suggests Cry toxin resistance may be induced in invertebrates as an active immune response. The use of model invertebrates, such as Caenorhabditis elegans and Drosophila melanogaster, as well as advances in insect genomics, are likely to accelerate efforts to clone Cry toxin resistance genes and come to a detailed and broad understanding of Cry toxin resistance.
TL;DR: Data indicate that long‐term exposure to insecticidal toxins occurs in the field, and levels of exposure should be considered during future risk assessments of transgenic crops to nontarget herbivores and arthropod predators.
Abstract: The planting of transgenic crops expressing Bacillus thuringiensis endotoxins is widespread throughout the world; the prolific increase in their application exposes nontarget organisms to toxins designed to control pests. To date, studies have focused upon the effects of Bt endotoxins on specific herbivores and detritivores, without consideration of their persistence within arthropod food webs. Here, we report the first quantitative field evaluation of levels of Bt endotoxin within nontarget herbivores and the uptake by higher order arthropods. Antibody-based assays indicated significant quantities of detectable Cry1Ab endotoxin within nontarget herbivores which feed on transgenic corn (including the corn flea beetle, Chaetocnema pulicaria, Japanese beetle, Popillia japonica and southern corn rootworm, Diabrotica undecimpunctata howardi ). Furthermore, arthropod predators (Coccinellidae, Araneae, and Nabidae) collected from these agroecosystems also contained significant quantities of Cry1Ab endotoxin indicating its movement into higher trophic levels. This uptake by predators is likely to have occurred by direct feeding on plant material (in predators which are facultatively phytophagous) or the consumption of arthropod prey which contained these proteins. These data indicate that long-term exposure to insecticidal toxins occurs in the field. These levels of exposure should therefore be considered during future risk assessments of transgenic crops to nontarget herbivores and arthropod predators.
TL;DR: It is found that the developmental decline in bioefficacy in field-grown plants was associated with reduced cry1Ac transcript levels and Bt toxin levels in postsquaring cotton, and changes in plant chemistry associated with the maturation of the cotton plant were observed to contribute to changes in the efficacy of Bt toxins.
Abstract: The insertion of genes encoding insecticidal Cry1A delta-endotoxins from the bacterium Bacillus thuringiensis Berliner variety kurstaki (Bt) into varieties of cotton (Gossypium spp.) was undertaken to assist in the control of a range of lepidopteran pests. In Australia, where Helicoverpa armigera (Hubner) and Helicoverpa punctigera (Wallengren) are major pests, the level of control is useful, but not complete, because efficacy declines as the crop matures. Fluctuations in the efficacy of Bt cotton, to the extent that some insects survive, provide opportunities for H. armigera to develop resistance to the Bt toxin. Therefore, variations in the efficacy of Bt cotton need to be understood if we are to plan rational resistance management strategies to retard the rate of the development of resistance. We measured the changes in efficacy associated with plant development over the growing season, in the field and glasshouse. In addition, the levels of Cry1Ac protein toxin and cry1Ac RNA were determined. In this first demonstration of the relationship between these three factors, we found that the developmental decline in bioefficacy in field-grown plants was associated with reduced cry1Ac transcript levels and Bt toxin levels in postsquaring cotton. In addition, changes in plant chemistry associated with the maturation of the cotton plant were observed to contribute to changes in the efficacy of Bt toxin. Results from the field and glasshouse suggested that variations in efficacy within the growing season and between seasons also may be influenced by environmental factors.
TL;DR: Combination of these physical and physiological conditions during agroinoculation, co-cultivation and selection on kanamycin were found to be critical determinants resulting into increased competence for regeneration and co-transformation frequency of cry1Ac and npt II and elimination of escapes.
TL;DR: Among 48,901 samples of ready-to-eat food products at the Danish retail market, 0.5% had counts of Bacillus cereus-like bacteria above 10(4) cfu g(-1) that may belong to B. thuringiensis, which is commonly found in starchy, cooked products, but also in fresh cucumbers and tomatoes.
Abstract: Among 48,901 samples of ready-to-eat food products at the Danish retail market, 0.5% had counts of Bacillus cereus-like bacteria above 10(4) cfu g(-1). The high counts were most frequently found in starchy, cooked products, but also in fresh cucumbers and tomatoes. Forty randomly selected strains had at least one gene or component involved in human diarrhoeal disease, while emetic toxin was related to only one B. cereus strain. A new observation was that 31 out of the 40 randomly selected B. cereus-like strains could be classified as Bacillus thuringiensis due to crystal production and/or content of cry genes. Thus, a large proportion of the B. cereus-like organisms present in food may belong to B. thuringiensis.
TL;DR: The effect of the Bt maize was small and within the normal variation expected in these agricultural systems, with one occurrence of CLPP varying between maize cultivars (Bt versus a conventional cultivar).
Abstract: Field trials were established at three European sites (Denmark, Eastern France, South-West France) of genetically modified maize (Zea mays L.) expressing the CryIAb Bacillus thuringiensis toxin (Bt), the near-isogenic non-Bt cultivar, another conventional maize cultivar and grass. Soil from Denmark was sampled at sowing (May) and harvest (October) over two years (2002, 2003); from E France at harvest 2002, sowing and harvest 2003; and from SW France at sowing and harvest 2003. Samples were analysed for microbial community structure (2003 samples only) by community-level physiological-profiling (CLPP) and phospholipid fatty acid analysis (PLFA), and protozoa and nematodes in all samples. Individual differences within a site resulted from: greater nematode numbers under grass than maize on three occasions; different nematode populations under the conventional maize cultivars once; and two occasions when there was a reduced protozoan population under Bt maize compared to non-Bt maize. Microbial community structure within the sites only varied with grass compared to maize, with one occurrence of CLPP varying between maize cultivars (Bt versus a conventional cultivar). An overall comparison of Bt versus non-Bt maize across all three sites only revealed differences for nematodes, with a smaller population under the Bt maize. Nematode community structure was different at each site and the Bt effect was not confined to specific nematode taxa. The effect of the Bt maize was small and within the normal variation expected in these agricultural systems.
TL;DR: Results indicate that B. thuringiensis and B. bassiana have strong potential for integrated biologically based management of Colorado potato beetle.
TL;DR: It is compared by 2‐DE in the early stationary phase of growth the extracellular proteomes of three strains of these species that have lost their virulence plasmids, finding large amounts of secreted degradative enzymes and toxins, including nine proteases, three phospholipases, two haemolysins and several enterotoxins.
Abstract: Bacillus cereus, Bacillus thuringiensis and Bacillus anthracis are closely related species that share a similar genetic background but occupy different ecological niches. Virulence plasmids bearing genes coding for toxins, may explain, at least partly, this specialization. We have compared by 2-DE in the early stationary phase of growth the extracellular proteomes of three strains of these species that have lost their virulence plasmids. Proteins expected to be secreted or to belong to the cell wall or to the cytosol were found in the three proteomes. For the cell wall and cytosolic proteins located in the extracellular space, the three proteomes were similar. Cytosolic proteins included enolase, GroEL, PdhB, PdhD, SodA and others. Cell surface proteins were mainly autolysins, proteases, nucleotidases and OppAs. In contrast, the secreted proteins profiles of B. cereus and B. thuringiensis were quite different from that of B. anthracis. B. cereus and B. thuringiensis extracellular proteomes both contained large amounts of secreted degradative enzymes and toxins, including nine proteases, three phospholipases, two haemolysins and several enterotoxins. Most of the genes encoding these enzymes and toxins are controlled by the transcriptional activator PlcR. The extracellular proteome of the pXO1-, pXO2- B. anthracis 9131 strain contained only one secreted protein: the metalloprotease InhA1, also found in the proteomes of the two other strains and possibly involved in antibacterial peptide degradation.
TL;DR: This work proposes a resistance mechanism against toxins based on a systemic immune-induction that can be transmitted to the next generation by a maternal effect and revealed elevated immune responses involving increased melanization and the presence of a soluble toxin-binding glycoprotein in the hemolymph and gut lumen of the resistant strain.
TL;DR: This article describes an alternative transgenic strategy that has the potential to generate broader and more sustainable levels of resistance against insect pests, and involves engineering plants with a fusion protein combining the δ-endotoxin Cry1Ac with the galactose-binding domain of the nontoxic ricin B-chain (RB).
Abstract: Bacillus thuringiensis (Bt) crystal protein genes encode insecticidal δ-endotoxins that are widely used for the development of insect-resistant crops. In this article, we describe an alternative transgenic strategy that has the potential to generate broader and more sustainable levels of resistance against insect pests. Our strategy involves engineering plants with a fusion protein combining the δ-endotoxin Cry1Ac with the galactose-binding domain of the nontoxic ricin B-chain (RB). This fusion, designated BtRB, provides the toxin with additional, binding domains, thus increasing the potential number of interactions at the molecular level in target insects. Transgenic rice and maize plants engineered to express the fusion protein were significantly more toxic in insect bioassays than those containing the Bt gene alone. They were also resistant to a wider range of insects, including important pests that are not normally susceptible to Bt toxins. The potential impact of fusion genes such as BtRB in terms of crop improvement, resistance sustainability, and biosafety is discussed.
TL;DR: The damage inflicted by cotton bollworm on cotton, regardless of the presence or absence of insecticidal genes, is predicted to be more serious under elevated CO2 conditions because of individual compensatory feeding on host plants caused by nitrogen deficiency.
Abstract: Effects of elevated atmospheric CO 2 (double-ambient CO 2 ) on the growth and metabolism of cotton bollworm, Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae), fed on transgenic Bacillus thuringiensis (Berliner) (Bt) cotton [CrylA(c)], grown in open-top chambers, were studied. Two levels of CO 2 (ambient and double-ambient) and two cotton cultivars (non-transgenic Simian-3 and transgenic GK-12) were deployed in a completely randomized design with four treatment combinations, and the cotton bollworm was reared on each treatment simultaneously. Plants of both cotton cultivars had lower nitrogen and higher total non-structural carbohydrates (TNC), TNC:Nitrogen ratio, condensed tannin, and gossypol under elevated CO 2 . Elevated CO 2 further resulted in a significant decrease in Bt toxin level in GK-12. The changes in chemical components in the host plants due to increased CO 2 significantly affected the growth parameters of H. armigera. Both transgenic Bt cotton and elevated CO 2 resulted in a reduced body mass, lower fecundity, decreased relative growth rate (RGR), and decreased mean relative growth rate in the bollworms. Larval life-span was significantly longer for H. armigera fed transgenic Bt cotton. Significantly reduced larval, pupal, and adult moth weights were observed in the bollworms fed elevated CO 2 -grown transgenic Bt cotton compared with those of bollworms reared on non-transgenic cotton, regardless of the CO 2 level. The efficiency of conversion of ingested food and of digested food of the bollworm were significantly reduced when fed transgenic Bt cotton, but there was no significant CO 2 or CO 2 x cotton cultivar interaction. Approximate digestibility of larvae reared on transgenic cotton grown in elevated CO 2 was higher compared to that of larvae fed non-transgenic cotton grown at ambient CO 2 . The damage inflicted by cotton bollworm on cotton, regardless of the presence or absence of insecticidal genes, is predicted to be more serious under elevated CO 2 conditions because of individual compensatory feeding on host plants caused by nitrogen deficiency.
TL;DR: The results showed that the toxin content in Bt cotton changed significantly over time, and that the structure, growth stage, and variety were significant sources of variability.
Abstract: Seasonal levels of Bacillus thuringiensis (Bt) insecticidal protein and its control efficacy against Helicoverpa armigera (Hubner) in Bt transgenic cotton GK19 (carrying a Cry1Ac/Cry1Ab fused gene) and BG1560 (carrying a Cry1Ac gene) were investigated in Tianmen County, Hubei Province, located in the Yangtze River valley of China, in 2001 and 2002. The results showed that the toxin content in Bt cotton changed significantly over time, and that the structure, growth stage, and variety were significant sources of variability. Generally, insecticidal protein levels were high during the early stages of cotton growth; they declined in mid-season, and rebounded in late season. On most dates sampled, the toxin contents in leaf, square, petal, and stamens (including nonovule pistil tissue) were much higher than those in ovule and boll. Compared with BG1560, the expression of Cry1Ac/Cry1Ab protein in GK19 was more variable during the whole growth period of cotton. The field evaluation on larval population dynamics of H. armigera in Bt and conventional cotton showed that the larval densities in BG1560 and GK19 fields decreased, respectively, 92.04 and 81.85% in 2001, and 96.84 and 91.80% in 2002.
TL;DR: Long-term studies in Arizona showed essentially no effects of Bt cotton on natural enemy function in Arizona cotton and showed that minor reductions in density of several predator taxa in Btotton observed in a companion study may have little ecological meaning relative to natural enemy impact on key pests in the system.
Abstract: Field studies to assess effects of transgenic crops on nontarget organisms primarily have focused on nontarget abundance and diversity, whereas few have examined effects on nontarget community function. A 5-yr field study in Arizona assessed whether transgenic cotton producing the Cry1Ac toxin of Bacillus thuringiensis (Bt) had an effect on the natural enemy community’s impact on three key pests. Predator:prey ratios for Bemisia tabaci (Gennadius) and Lygus hesperus Knight were similar in unsprayed Bt and non-Bt cotton. Insecticide applications in positive control plots inconsistently altered ratios for B. tabaci. Predation indices based on the known feeding activity of selected predators showed that potential predator impact was unaltered by Bt cotton but was consistently depressed with insecticide applications. Sentinel eggs and pupae of Pectinophora gossypiella (Saunders) experienced the same rates of predation in both unsprayed Bt and non-Bt cotton. Cohort-based life tables for B. tabaci show...
TL;DR: It is suggested that the observed susceptibility differences reflect natural variation in Bt susceptibility among rootworm populations and provide a baseline for estimating potential shifts in susceptibility that might result from selection and exposure to Cry3Bb1-expressing corn hybrids.
Abstract: Susceptibility to Cry3Bb1 toxin from Bacillus thuringiensis (Bt) was determined for western corn rootworm, Diabrotica virgifera virgifera LeConte, neonates from both laboratory and field populations collected from across the Corn Belt. Rootworm larvae were exposed to artificial diet treated with increasing Cry3Bb1 concentrations, and mortality and growth inhibition were evaluated after 4–7 d. The range of variation in Bt susceptibility indicated by growth inhibition was similar to that indicated by mortality. Although interpopulation variation in susceptibility was observed, the magnitude of the differences was comparable with the variability observed between generations of the same population. In general, the toxin was not highly toxic to larvae and estimated LC50 and EC50 values were several times higher than those reported for lepidopteran-specific Cry toxins by using similar bioassay techniques. These results suggest that the observed susceptibility differences reflect natural variation in Bt susceptibility among rootworm populations and provide a baseline for estimating potential shifts in susceptibility that might result from selection and exposure to Cry3Bb1-expressing corn hybrids.