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

机译:来自棉铃虫田间种群的突变钙黏着蛋白等位基因赋予对苏云金芽孢杆菌毒素Cry1Ac▿的抗性

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摘要

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 (r1) 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 (r2 and r3) associated with Cry1Ac resistance from a field population of H. armigera collected from the Yellow River cotton area of China in 2005. The r2 and r3 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.
机译:棉铃虫棉铃虫(Helicoverpa armigera)是转基因棉花所针对的主要昆虫,其基因工程化生产了旧世界的苏云金芽孢杆菌毒素(转基因Bt棉花)。这种有害生物对苏云金芽孢杆菌毒素的抗性演变是转基因Bt棉花长期有效性的主要威胁。钙粘蛋白基因(Ha_BtR)的缺失突变等位基因(r1)先前被确定为与实验室选择的棉铃虫菌株中的Cry1Ac抗性遗传相关。使用双相筛选策略,我们从2005年中国黄河棉区的棉铃虫田间种群中成功捕获了两个与Cry1Ac抗性相关的钙黏着蛋白新等位基因(r2和r3)。r2和r3等位基因分别为通过在Ha_BtR的第8外显子的相同位置插入反转录转座子(称为HaRT1)的长末端重复序列和完整的HaRT1反转录转座子,产生了截短的钙粘着蛋白,该截短的钙粘着蛋白在Ha_BtR的N末端仅包含两个胞外域重复序列。这是首次在野外成功检测到Bt作物目标昆虫的苏云金芽孢杆菌抗性等位基因。这项研究还表明,带有两个抗性等位基因的棉铃虫幼虫可以在Bt棉上完成发育。钙粘蛋白基因座应该是对棉铃虫田间种群进行基于DNA密集筛选的重要目标。

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