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苏云金芽孢杆菌vip3BR杀虫毒素基因的缺失突变体ndv200是下一代抗鳞翅目昆虫危害的转基因作物的潜在候选基因。

A deletion mutant ndv200 of the Bacillus thuringiensis vip3BR insecticidal toxin gene is a prospective candidate for the next generation of genetically modified crop plants resistant to lepidopteran insect damage.

作者信息

Gayen Srimonta, Samanta Milan Kumar, Hossain Munshi Azad, Mandal Chandi Charan, Sen Soumitra Kumar

机构信息

Department of Human Genetics, University of Michigan, Ann Arbor, MI, 48108, USA,

出版信息

Planta. 2015 Jul;242(1):269-81. doi: 10.1007/s00425-015-2309-1. Epub 2015 Apr 26.

Abstract

Ectopic expression of a deletion mutant ( ndv200 ) of Bacillus thuringiensis vip3BR gene in tobacco plant provided almost complete protection against major crop pests cotton boll worm ( Helicoverpa armigera ), black cut worm ( Agrotis ipsilon ) and cotton leaf worm ( Spodoptera littoralis ). Whereas vip3BR transgenic tobacco plant failed to protect themselves from these insects and showed resistance towards cotton leaf worm only. An analogous form of the Bacillus thuringiensis vip3Aa insecticidal toxin gene, named vip3BR, was identified and characterized, and exhibited similar attributes to the well-known Vip3Aa toxin. Vip3BR possessed broad-spectrum lepidopteran-specific insecticidal properties effective against most major crop pests of the Indian subcontinent. A Vip3BR toxin protein N-terminal deletion mutant, Ndv200, showed increased insecticidal potency relative to the native toxin, which conferred efficacy against four major crop pests, including cotton boll worm (Helicoverpa armigera), black cut worm (Agrotis ipsilon), cotton leaf worm (Spodoptera littoralis), and rice yellow stem borer (Scirpophaga incertulas). Ligand blot analysis indicated the Ndv200 toxin recognized the same larval midgut receptors as the native Vip3BR toxin, but differed from receptors recognized by Cry1A toxins. In the present study, we tested the prospect of the vip3BR and ndv200 toxin gene as candidate in development of insect-resistant genetically engineered crop plants by generating transgenic tobacco plant. The study revealed that the ndv200 mutant of vip3BR insecticidal toxin gene is a strong and prospective candidate for the next generation of genetically modified crop plants resistant to lepidopteran insects.

摘要

苏云金芽孢杆菌vip3BR基因缺失突变体(ndv200)在烟草植株中的异位表达为主要农作物害虫棉铃虫(棉铃实夜蛾)、小地老虎(小地老虎)和棉叶虫(滨海灰翅夜蛾)提供了几乎完全的保护。而vip3BR转基因烟草植株无法保护自身免受这些昆虫侵害,仅对棉叶虫表现出抗性。一种名为vip3BR的苏云金芽孢杆菌vip3Aa杀虫毒素基因的类似形式被鉴定和表征,并表现出与著名的Vip3Aa毒素相似的特性。Vip3BR具有广谱鳞翅目特异性杀虫特性,对印度次大陆的大多数主要农作物害虫有效。一种Vip3BR毒素蛋白N端缺失突变体Ndv200相对于天然毒素显示出更高的杀虫效力,它对四种主要农作物害虫具有效力,包括棉铃虫(棉铃实夜蛾)、小地老虎(小地老虎)、棉叶虫(滨海灰翅夜蛾)和水稻二化螟(三化螟)。配体印迹分析表明,Ndv200毒素识别的幼虫中肠受体与天然Vip3BR毒素相同,但与Cry1A毒素识别的受体不同。在本研究中,我们通过培育转基因烟草植株,测试了vip3BR和ndv200毒素基因作为抗虫基因工程作物植物开发候选基因的前景。研究表明,vip3BR杀虫毒素基因的ndv200突变体是下一代抗鳞翅目昆虫转基因作物植物的强大且有前景的候选基因。

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