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一种新型ABC转运蛋白基因(Pxwhite)的下调与小菜蛾(Plutella xylostella (L.))对Cry1Ac的抗性相关。

Down-regulation of a novel ABC transporter gene (Pxwhite) is associated with Cry1Ac resistance in the diamondback moth, Plutella xylostella (L.).

作者信息

Guo Zhaojiang, Kang Shi, Zhu Xun, Xia Jixing, 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

Insect Biochem Mol Biol. 2015 Apr;59:30-40. doi: 10.1016/j.ibmb.2015.01.009. Epub 2015 Jan 27.

Abstract

Biopesticides or transgenic crops based on Cry toxins from the soil bacterium Bacillus thuringiensis (Bt) effectively control agricultural insect pests. The sustainable use of Bt biopesticides and Bt crops is threatened, however, by the development of Cry resistance in the target pests. The diamondback moth, Plutella xylostella (L.), is the first pest that developed resistance to a Bt biopesticide in the field, and a recent study has shown that the resistance of P. xylostella to Cry1Ac is caused by a mutation in an ATP-binding cassette (ABC) transporter gene (ABCC2). In this study, we report that down-regulation of a novel ABC transporter gene from ABCG subfamily (Pxwhite) is associated with Cry1Ac resistance in P. xylostella. The full-length cDNA sequence of Pxwhite was cloned and analyzed. Spatial-temporal expression detection revealed that Pxwhite was expressed in all tissues and developmental stages, and highest expressed in Malpighian tubule tissue and in egg stage. Sequence variation analysis of Pxwhite indicated the absence of constant non-synonymous mutations between susceptible and resistant strains, whereas midgut transcript analysis showed that Pxwhite was remarkably reduced in all resistant strains and further reduced when larvae of the moderately resistant SZ-R strain were subjected to selection with Cry1Ac toxin. Furthermore, RNA interference (RNAi)-mediated suppression of Pxwhite gene expression significantly reduced larval susceptibility to Cry1Ac toxin, and genetic linkage analysis confirmed that down-regulation of Pxwhite gene is tightly linked to Cry1Ac resistance in P. xylostella. To our knowledge, this is the first report indicating that Pxwhite gene is involved in Cry1Ac resistance in P. xylostella.

摘要

基于土壤细菌苏云金芽孢杆菌(Bt)的Cry毒素的生物农药或转基因作物能有效防治农业害虫。然而,目标害虫对Cry毒素产生抗性的发展威胁着Bt生物农药和Bt作物的可持续利用。小菜蛾(Plutella xylostella (L.))是田间第一种对Bt生物农药产生抗性的害虫,最近的一项研究表明,小菜蛾对Cry1Ac的抗性是由ATP结合盒(ABC)转运蛋白基因(ABCC2)的突变引起的。在本研究中,我们报告了ABCG亚家族的一个新的ABC转运蛋白基因(Pxwhite)的下调与小菜蛾对Cry1Ac的抗性有关。克隆并分析了Pxwhite的全长cDNA序列。时空表达检测显示,Pxwhite在所有组织和发育阶段均有表达,在马氏管组织和卵期表达最高。Pxwhite的序列变异分析表明,敏感和抗性品系之间不存在恒定的非同义突变,而中肠转录本分析表明,所有抗性品系中Pxwhite均显著降低,当中等抗性的SZ-R品系幼虫用Cry1Ac毒素进行选择时,Pxwhite进一步降低。此外,RNA干扰(RNAi)介导的Pxwhite基因表达抑制显著降低了幼虫对Cry1Ac毒素的敏感性,遗传连锁分析证实,Pxwhite基因的下调与小菜蛾对Cry1Ac的抗性紧密相关。据我们所知,这是第一份表明Pxwhite基因参与小菜蛾对Cry1Ac抗性的报告。

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