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小菜蛾中肠类钙黏蛋白基因与小菜蛾对苏云金芽孢杆菌毒素Cry1Ac的抗性无关。

The midgut cadherin-like gene is not associated with resistance to Bacillus thuringiensis toxin Cry1Ac in Plutella xylostella (L.).

作者信息

Guo Zhaojiang, Kang Shi, Zhu Xun, Wu Qingjun, Wang Shaoli, Xie Wen, Zhang Youjun

机构信息

Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China.

出版信息

J Invertebr Pathol. 2015 Mar;126:21-30. doi: 10.1016/j.jip.2015.01.004. Epub 2015 Jan 13.

Abstract

The Gram-positive bacterium Bacillus thuringiensis (Bt) produces Cry toxins that have been used to control important agricultural pests. Evolution of resistance in target pests threatens the effectiveness of these toxins when used either in sprayed biopesticides or in Bt transgenic crops. Although alterations of the midgut cadherin-like receptor can lead to Bt Cry toxin resistance in many insects, whether the cadherin gene is involved in Cry1Ac resistance of Plutella xylostella (L.) remains unclear. Here, we present experimental evidence that resistance to Cry1Ac or Bt var. kurstaki (Btk) in P. xylostella is not due to alterations of the cadherin gene. The bona fide P. xylostella cadherin cDNA sequence was cloned and analyzed, and comparisons of the cadherin cDNA sequence among susceptible and resistant P. xylostella strains confirmed that Cry1Ac resistance was independent of mutations in this gene. In addition, real-time quantitative PCR (qPCR) indicated that cadherin transcript levels did not significantly differ among susceptible and resistant P. xylostella strains. RNA interference (RNAi)-mediated suppression of cadherin gene expression did not affect larval susceptibility to Cry1Ac toxin. Furthermore, genetic linkage assays using four cadherin gDNA allelic biomarkers confirmed that the cadherin gene is not linked to resistance against Cry1Ac in P. xylostella. Taken together, our findings demonstrate that Cry1Ac resistance of P. xylostella is independent of the cadherin gene.

摘要

革兰氏阳性细菌苏云金芽孢杆菌(Bt)产生的Cry毒素已被用于控制重要的农业害虫。目标害虫产生抗性会威胁到这些毒素在喷雾生物农药或Bt转基因作物中的使用效果。尽管中肠钙黏蛋白样受体的改变会导致许多昆虫对Bt Cry毒素产生抗性,但钙黏蛋白基因是否参与小菜蛾对Cry1Ac的抗性仍不清楚。在此,我们提供实验证据表明,小菜蛾对Cry1Ac或Bt库斯塔克变种(Btk)的抗性并非由于钙黏蛋白基因的改变。我们克隆并分析了小菜蛾真正的钙黏蛋白cDNA序列,对敏感和抗性小菜蛾品系的钙黏蛋白cDNA序列进行比较,证实Cry1Ac抗性与该基因的突变无关。此外,实时定量PCR(qPCR)表明,敏感和抗性小菜蛾品系之间的钙黏蛋白转录水平没有显著差异。RNA干扰(RNAi)介导的钙黏蛋白基因表达抑制并未影响幼虫对Cry1Ac毒素的敏感性。此外,使用四个钙黏蛋白gDNA等位生物标志物进行的遗传连锁分析证实,钙黏蛋白基因与小菜蛾对Cry1Ac的抗性不相关。综上所述,我们的研究结果表明,小菜蛾对Cry1Ac的抗性与钙黏蛋白基因无关。

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