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基于DNA的棉铃虫田间种群细胞内钙黏蛋白突变检测,该突变赋予对Cry1Ac非隐性抗性

DNA-based screening for an intracellular cadherin mutation conferring non-recessive Cry1Ac resistance in field populations of Helicoverpa armigera.

作者信息

Zhang Haonan, Tang Mingyi, Yang Fan, Yang Yihua, Wu Yidong

机构信息

Key Laboratory of Integrated Management of Crop Diseases and Pests (Ministry of Education), College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.

Key Laboratory of Integrated Management of Crop Diseases and Pests (Ministry of Education), College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.

出版信息

Pestic Biochem Physiol. 2013 Sep;107(1):148-52. doi: 10.1016/j.pestbp.2013.06.007. Epub 2013 Jul 2.

DOI:10.1016/j.pestbp.2013.06.007
PMID:25149249
Abstract

A number of cadherin mutants conferring resistance to Bt toxin Cry1Ac have been reported in three major lepidopteran pests, including Helicoverpa armigera. Unlike most of the cadherin mutants conferring recessive resistance, an allele (r15) with a 55aa deletion in the intracellular domain of cadherin (HaCad) was previously identified to cause non-recessive resistance to Cry1Ac in H. armigera. In the present study, a DNA-based PCR method was developed to screen the r15 allele from field populations of H. armigera collected from the main cotton planting areas of China in 2011 and 2012. Three heterozygous r15 alleles were detected from 562 moths collected from northern China (with intensive Bt cotton planting), and r15 allele frequency was estimated to be 0.0027. However, no r15 allele was detected from 314 moths collected from Xinjiang (with limited Bt cotton use). Although all the r15 alleles have the same deletion in the cDNA sequence, at least four different indels causing loss of exon 32 have been detected in the genomic DNA sequences flanking exon 32 of HaCad. Multiple origins of the r15 alleles illustrate parallel genotypic adaption of H. armigera to the selection pressure of Bt cotton.

摘要

在包括棉铃虫在内的三种主要鳞翅目害虫中,已报道了一些对Bt毒素Cry1Ac具有抗性的钙黏蛋白突变体。与大多数赋予隐性抗性的钙黏蛋白突变体不同,先前已鉴定出一种在钙黏蛋白(HaCad)细胞内结构域中有55个氨基酸缺失的等位基因(r15),它会导致棉铃虫对Cry1Ac产生非隐性抗性。在本研究中,开发了一种基于DNA的PCR方法,用于从2011年和2012年从中国主要棉花种植区采集的棉铃虫田间种群中筛选r15等位基因。从中国北方(Bt棉花种植密集)采集的562只蛾子中检测到三个杂合r15等位基因,并估计r15等位基因频率为0.0027。然而,从新疆(Bt棉花使用有限)采集的314只蛾子中未检测到r15等位基因。尽管所有r15等位基因在cDNA序列中具有相同的缺失,但在HaCad第32外显子侧翼的基因组DNA序列中至少检测到四种不同的导致第32外显子缺失的插入缺失。r15等位基因的多个起源说明了棉铃虫对Bt棉花选择压力的平行基因型适应。

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