Yang Yajun, Chen Haiyan, Wu Yidong, Yang Yihua, Wu Shuwen
Department of Entomology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.
Appl Environ Microbiol. 2007 Nov;73(21):6939-44. doi: 10.1128/AEM.01703-07. Epub 2007 Sep 7.
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.
棉铃虫是旧大陆上转苏云金芽孢杆菌毒素基因棉花(转基因Bt棉花)所针对的主要害虫。这种害虫对苏云金芽孢杆菌毒素产生抗性的进化是转基因Bt棉花长期有效性的主要威胁。先前在实验室筛选的棉铃虫品系中,一个钙黏蛋白基因(Ha_BtR)的缺失突变等位基因(r(1))被确定与对Cry1Ac的抗性存在遗传连锁。我们采用双相筛选策略,成功地从2005年采集自中国黄河棉区的棉铃虫田间种群中捕获到两个与Cry1Ac抗性相关的新钙黏蛋白等位基因(r(2)和r(3))。r(2)和r(3)等位基因分别是通过将一个反转录转座子(命名为HaRT1)的长末端重复序列和完整的HaRT1反转录转座子插入到Ha_BtR外显子8的同一位置而产生的,这导致Ha_BtR的N端仅含有两个胞外结构域重复序列的截短型钙黏蛋白。这是首次在野外成功检测到Bt作物目标害虫的苏云金芽孢杆菌抗性等位基因。本研究还表明,携带两个抗性等位基因的棉铃虫幼虫能够在Bt棉花上完成发育。钙黏蛋白基因座应成为对棉铃虫田间种群进行基于DNA的密集筛选的重要靶点。