scholarly journals Interaction of Bacillus thuringiensis Toxins with Larval Midgut Binding Sites of Helicoverpa armigera (Lepidoptera: Noctuidae)

2004 ◽  
Vol 70 (3) ◽  
pp. 1378-1384 ◽  
Author(s):  
Anna Estela ◽  
Baltasar Escriche ◽  
Juan Ferr�
Keyword(s):  
Bacillus Thuringiensis ◽  
Concanavalin A ◽  
Helicoverpa Armigera ◽  
Binding Sites ◽  
Inhibitory Effect ◽  
Toxin Gene ◽  
Bt Cotton ◽  
Cry1ab Protein ◽  
Cry Proteins ◽  
Helicoverpa Armígera

ABSTRACT In 1996, Bt-cotton (cotton expressing a Bacillus thuringiensis toxin gene) expressing the Cry1Ac protein was commercially introduced to control cotton pests. A threat to this first generation of transgenic cotton is the evolution of resistance by the insects. Second-generation Bt-cotton has been developed with either new B. thuringiensis genes or with a combination of cry genes. However, one requirement for the “stacked” gene strategy to work is that the stacked toxins bind to different binding sites. In the present study, the binding of 125I-labeled Cry1Ab protein (125I-Cry1Ab) and 125I-Cry1Ac to brush border membrane vesicles (BBMV) of Helicoverpa armigera was analyzed in competition experiments with 11 nonlabeled Cry proteins. The results indicate that Cry1Aa, Cry1Ab, and Cry1Ac competed for common binding sites. No other Cry proteins tested competed for either 125I-Cry1Ab or 125I-Cry1Ac binding, except Cry1Ja, which competed only at the highest concentrations used. Furthermore, BBMV from four H. armigera populations were also tested with 125I-Cry1Ac and Cry1Ab to check the influence of the insect population on the binding results. Finally, the inhibitory effect of selected sugars and lectins was also determined. 125I-Cry1Ac binding was strongly inhibited by N-acetylgalactosamine, sialic acid, and concanavalin A and moderately inhibited by soybean agglutinin. In contrast, 125I-Cry1Ab binding was only significantly inhibited by concanavalin A. These results show that Cry1Ac and Cry1Ab use different epitopes for binding to BBMV.

scholarly journals Disruption of a Cadherin Gene Associated with Resistance to Cry1Ac δ-Endotoxin of Bacillus thuringiensis in Helicoverpa armigera

2005 ◽  
Vol 71 (2) ◽  
pp. 948-954 ◽  
Author(s):  
Xinjun Xu ◽  
Liangying Yu ◽  
Yidong Wu
Keyword(s):  
Bacillus Thuringiensis ◽  
Helicoverpa Armigera ◽  
Stop Codon ◽  
Premature Stop Codon ◽  
Recessive Gene ◽  
Laboratory Strain ◽  
Cross Resistance ◽  
Resistance Pattern ◽  
Bt Cotton ◽  
Helicoverpa Armígera

ABSTRACT A laboratory strain (GY) of Helicoverpa armigera (Hübner) 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.

scholarly journals Effect of NaCl-stressed Bacillus thuringiensis (Bt) cotton on the feeding behaviors and nutritional parameters of Helicoverpa armigera

PLoS ONE ◽  
2018 ◽  
Vol 13 (9) ◽  
pp. e0198570
Author(s):  
Jun-Yu Luo ◽  
Shuai Zhang ◽  
Xiang-Zhen Zhu ◽  
Ji-Chao Ji ◽  
Kai-Xin Zhang ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Helicoverpa Armigera ◽  
Bt Cotton ◽  
Feeding Behaviors ◽  
Nutritional Parameters ◽  
Helicoverpa Armígera

scholarly journals Mutated Cadherin Alleles from a Field Population of Helicoverpa armigera Confer Resistance to Bacillus thuringiensis Toxin Cry1Ac

2007 ◽  
Vol 73 (21) ◽  
pp. 6939-6944 ◽  
Author(s):  
Yajun Yang ◽  
Haiyan Chen ◽  
Yidong Wu ◽  
Yihua Yang ◽  
Shuwen Wu
Keyword(s):  
Bacillus Thuringiensis ◽  
Helicoverpa Armigera ◽  
Yellow River ◽  
Insect Pest ◽  
Genetically Engineered ◽  
Field Population ◽  
Bt Cotton ◽  
Transgenic Bt Cotton ◽  
Helicoverpa Armígera ◽  
First Time

ABSTRACT The cotton bollworm Helicoverpa armigera is the major insect pest targeted by cotton genetically engineered to produce the Bacillus thuringiensis toxin (transgenic Bt cotton) in the Old World. The evolution of this pest's resistance to B. thuringiensis toxins is the main threat to the long-term effectiveness of transgenic Bt cotton. A deletion mutation allele (r 1 ) of a cadherin gene (Ha_BtR) was previously identified as genetically linked with Cry1Ac resistance in a laboratory-selected strain of H. armigera. Using a biphasic screen strategy, we successfully trapped two new cadherin alleles (r 2 and r 3 ) associated with Cry1Ac resistance from a field population of H. armigera collected from the Yellow River cotton area of China in 2005. The r 2 and r 3 alleles, respectively, were created by inserting the long terminal repeat of a retrotransposon (designated HaRT1) and the intact HaRT1 retrotransposon at the same position in exon 8 of Ha_BtR, which results in a truncated cadherin containing only two ectodomain repeats in the N terminus of Ha_BtR. This is the first time that the B. thuringiensis resistance alleles of a target insect of Bt crops have been successfully detected in the open field. This study also demonstrated that bollworm larvae carrying two resistance alleles can complete development on Bt cotton. The cadherin locus should be an important target for intensive DNA-based screening of field populations of H. armigera.

scholarly journals Effectiveness of Bacillus thuringiensis-Transgenic Chickpeas and the Entomopathogenic Fungus Metarhizium anisopliae in Controlling Helicoverpa armigera (Lepidoptera: Noctuidae)

2008 ◽  
Vol 74 (14) ◽  
pp. 4381-4389 ◽  
Author(s):  
N. C. Lawo ◽  
R. J. Mahon ◽  
R. J. Milner ◽  
B. K. Sarmah ◽  
T. J. V. Higgins ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Helicoverpa Armigera ◽  
Metarhizium Anisopliae ◽  
Entomopathogenic Fungus ◽  
Fitness Costs ◽  
Cry Proteins ◽  
Bt Crops ◽  
Helicoverpa Armígera ◽  
Lepidoptera Noctuidae ◽  
Increased Susceptibility

ABSTRACT The use of genetically modified (Bt) crops expressing lepidopteran-specific Cry proteins derived from the soil bacterium Bacillus thuringiensis is an effective method to control the polyphagous pest Helicoverpa armigera. As H. armigera potentially develops resistance to Cry proteins, Bt crops should be regarded as one tool in integrated pest management. Therefore, they should be compatible with biological control. Bioassays were conducted to understand the interactions between a Cry2Aa-expressing chickpea line, either a susceptible or a Cry2A-resistant H. armigera strain, and the entomopathogenic fungus Metarhizium anisopliae. In a first concentration-response assay, Cry2A-resistant larvae were more tolerant of M. anisopliae than susceptible larvae, while in a second bioassay, the fungus caused similar mortalities in the two strains fed control chickpea leaves. Thus, resistance to Cry2A did not cause any fitness costs that became visible as increased susceptibility to the fungus. On Bt chickpea leaves, susceptible H. armigera larvae were more sensitive to M. anisopliae than on control leaves. It appeared that sublethal damage induced by the B. thuringiensis toxin enhanced the effectiveness of M. anisopliae. For Cry2A-resistant larvae, the mortalities caused by the fungus were similar when they were fed either food source. To examine which strain would be more likely to be exposed to the fungus, their movements on control and Bt chickpea plants were compared. Movement did not appear to differ among larvae on Bt or conventional chickpeas, as indicated by the number of leaflets damaged per leaf. The findings suggest that Bt chickpeas and M. anisopliae are compatible to control H. armigera.

scholarly journals Need for growing non-Bt cotton refugia to overcome Bt resistance problem in targeted larvae of the cotton bollworms, Helicoverpa armigera and Pectinophora gossypiella

2021 ◽  
Vol 31 (1) ◽  
Author(s):  
Muhammad Rafiq Shahid ◽  
Muhammad Farooq ◽  
Muhammad Shakeel ◽  
Misbah Ashraf ◽  
Zia Ullah Zia ◽  
...  
Keyword(s):  
Helicoverpa Armigera ◽  
Larval Mortality ◽  
Pectinophora Gossypiella ◽  
Bt Cotton ◽  
Insect Larvae ◽  
Plant Parts ◽  
Arthropod Species ◽  
Bt Resistance ◽  
American Bollworm ◽  
Helicoverpa Armígera

Abstract Background The effectiveness of Bacillus thuringiensis (Bt) cotton against target arthropod larvae is decreasing day by day. The comparative effect of Bt expression among Bt cotton varieties and different plant parts was observed against the cotton bollworms: Helicoverpa armigera and Pectinophora gossypiella larvae. Results In the present study, larval mortality of H. armigera was higher than P. gossypiella among selected Bt cultivars. Median lethal concentration (LC50) values were 8.91, 13.4, 14.0, and 36.4 for P. gossypiella, while 5.91, 4.04, 2.37, and 8.26 for H. armigera of FH-142, MNH-886, IR-3701, and FH-Lalazar, respectively. These values depicted that P. gossypiella had more Bt resistance problem than H. armigera larvae. The host range of both targeted insect larvae was different from each other due to the polyphagous feeding nature of the larvae of H. armigera that feed on different host plants, but P. gossypiella attacked only cotton with monophagous feeding habit. It was also notable from results that Bt expression in reproductive parts where the attacked pink bollworm was lower than the American bollworm, so the former had the maximum chance of resistance due to repeated exposure to Bt. Conclusions It was concluded that farmers be advised to follow the practice of growing non-Bt as a refuge crop to reduce the problem of Bt resistance in the target arthropod species.

scholarly journals Bacillus thuringiensis Bel Protein Enhances the Toxicity of Cry1Ac Protein to Helicoverpa armigera Larvae by Degrading Insect Intestinal Mucin

2009 ◽  
Vol 75 (16) ◽  
pp. 5237-5243 ◽  
Author(s):  
Shangling Fang ◽  
Li Wang ◽  
Wei Guo ◽  
Xia Zhang ◽  
Donghai Peng ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Helicoverpa Armigera ◽  
Trichoplusia Ni ◽  
Knockout Mutant ◽  
Gene Encoding ◽  
In Vitro Degradation ◽  
Intestinal Mucin ◽  
Helicoverpa Armígera ◽  
Cry1ac Protein

ABSTRACT Bacillus thuringiensis has been used as a bioinsecticide to control agricultural insects. Bacillus cereus group genomes were found to have a Bacillus enhancin-like (bel) gene, encoding a peptide with 20 to 30% identity to viral enhancin protein, which can enhance viral infection by degradation of the peritrophic matrix (PM) of the insect midgut. In this study, the bel gene was found to have an activity similar to that of the viral enhancin gene. A bel knockout mutant was constructed by using a plasmid-free B. thuringiensis derivative, BMB171. The 50% lethal concentrations of this mutant plus the cry1Ac insecticidal protein gene were about 5.8-fold higher than those of the BMB171 strain. When purified Bel was mixed with the Cry1Ac protein and fed to Helicoverpa armigera larvae, 3 μg/ml Cry1Ac alone induced 34.2% mortality. Meanwhile, the mortality rate rose to 74.4% when the same amount of Cry1Ac was mixed with 0.8 μg/ml of Bel. Microscopic observation showed a significant disruption detected on the midgut PM of H. armigera larvae after they were fed Bel. In vitro degradation assays showed that Bel digested the intestinal mucin (IIM) of Trichoplusia ni and H. armigera larvae to various degrading products, similar to findings for viral enhancin. These results imply Bel toxicity enhancement depends on the destruction of midgut PM and IIM, similar to the case with viral enhancin. This discovery showed that Bel has the potential to enhance insecticidal activity of B. thuringiensis-based biopesticides and transgenic crops.

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