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Non-Recessive Bt Toxin Resistance Conferred by an Intracellular Cadherin Mutation in Field-Selected Populations of Cotton Bollworm

机译:田间选择棉铃虫的细胞内钙黏着蛋白突变赋予非隐性的Bt毒素抗性。

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摘要

Transgenic crops producing Bacillus thuringiensis (Bt) toxins have been planted widely to control insect pests, yet evolution of resistance by the pests can reduce the benefits of this approach. Recessive mutations in the extracellular domain of toxin-binding cadherin proteins that confer resistance to Bt toxin Cry1Ac by disrupting toxin binding have been reported previously in three major lepidopteran pests, including the cotton bollworm, Helicoverpa armigera. Here we report a novel allele from cotton bollworm with a deletion in the intracellular domain of cadherin that is genetically linked with non-recessive resistance to Cry1Ac. We discovered this allele in each of three field-selected populations we screened from northern China where Bt cotton producing Cry1Ac has been grown intensively. We expressed four types of cadherin alleles in heterologous cell cultures: susceptible, resistant with the intracellular domain mutation, and two complementary chimeric alleles with and without the mutation. Cells transfected with each of the four cadherin alleles bound Cry1Ac and were killed by Cry1Ac. However, relative to cells transfected with either the susceptible allele or the chimeric allele lacking the intracellular domain mutation, cells transfected with the resistant allele or the chimeric allele containing the intracellular domain mutation were less susceptible to Cry1Ac. These results suggest that the intracellular domain of cadherin is involved in post-binding events that affect toxicity of Cry1Ac. This evidence is consistent with the vital role of the intracellular region of cadherin proposed by the cell signaling model of the mode of action of Bt toxins. Considered together with previously reported data, the results suggest that both pore formation and cell signaling pathways contribute to the efficacy of Bt toxins.
机译:已经广泛种植了生产苏云金芽孢杆菌毒素的转基因作物,以控制害虫,但是害虫产生抗性的进化会降低这种方法的优势。先前已经在包括棉铃虫,棉铃虫在内的三种主要鳞翅目害虫中报道了毒素结合钙黏着蛋白蛋白胞外域的隐性突变,该蛋白通过破坏毒素结合而赋予对Bt毒素Cry1Ac的抗性。在这里,我们报道了棉铃虫的一个新的等位基因,在钙粘着蛋白的细胞内结构域中缺失,该基因在遗传上与对Cry1Ac的非隐性抗性相关。我们在从中国北方筛选出的三个田间选择种群中的每一个中发现了该等位基因,在中国北方,种植Bt棉的Cry1Ac已大量种植。我们在异源细胞培养物中表达了四种钙粘蛋白等位基因:易感性,对细胞内结构域突变具有抗性,以及两个互补的具有和不具有突变的嵌合等位基因。用四个钙粘蛋白等位基因的每一个转染的细胞结合Cry1Ac,并被Cry1Ac杀死。然而,相对于用易感的等位基因或缺少细胞内结构域突变的嵌合等位基因转染的细胞,用抗性等位基因或含有细胞内结构域突变的嵌合等位基因转染的细胞对Cry1Ac的敏感性较低。这些结果表明钙粘着蛋白的胞内域参与影响Cry1Ac毒性的结合后事件。该证据与Bt毒素作用方式的细胞信号传导模型提出的钙黏着蛋白细胞内区域的重要作用是一致的。与先前报道的数据一起考虑,结果表明孔形成和细胞信号通路均有助于Bt毒素的功效。

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