表达四泛素基因 HaTSPAN1 的 T92C 等位基因的基于 piggyBac 的转基因 Helicoverpa armigera 对苏云金芽孢杆菌毒素 Cry1Ac 具有显性抗性

IF 4.2 1区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Lin Li, Xinru Pang, Chenyang Wang, Yihua Yang, Yidong Wu
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引用次数: 0

摘要

生产苏云金芽孢杆菌(Bt)杀虫蛋白的转基因作物为害虫控制带来了革命性的变化。然而,目标害虫的抗药性进化对 Bt 作物的长期成功构成了重大威胁。了解 Bt 抗性的遗传学和机制对于开发抗性检测方法和管理策略至关重要。四泛素基因(HaTSPAN1)中的 T92C 突变导致 L31S 取代,与主要害虫 Helicoverpa armigera 对 Cry1Ac 的显性抗性有关。此前利用 CRISPR/Cas9 技术进行的研究表明,敲除 HaTSPAN1 T92C 突变基因可使对 Cry1Ac 产生抗性的 H. armigera SCD 易感株系的抗性提高 125 倍。在本研究中,我们利用 piggyBac 转座子系统创建了两个基于 SCD 的转基因 H. armigera 株系:一个表达野生型 HaTSPAN1 基因(SCD-TSPANwt),另一个表达 HaTSPAN1 的 T92C 突变体形式(SCD-TSPANmt)。与受体 SCD 菌株相比,SCD-TSPANmt 菌株对 Cry1Ac 的抗性提高了 82 倍,而 SCD-TSPANwt 菌株仍然易感。Cry1Ac抗性遵循常染色体显性遗传模式,并与SCD-TSPANmt株系中的H. armigera转基因位点有遗传联系。我们的研究结果进一步证实了 HaTSPAN1 的 T92C 突变与 H. armigera 对 Cry1Ac 的显性抗性之间的因果关系。此外,这些结果还表明,我们在蓟马中使用的 piggyBac 介导的转化系统有望用于其他鳞翅目害虫候选 Bt 抗性基因的功能研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

piggyBac-based transgenic Helicoverpa armigera expressing the T92C allele of the tetraspanin gene HaTSPAN1 confers dominant resistance to Bacillus thuringiensis toxin Cry1Ac

piggyBac-based transgenic Helicoverpa armigera expressing the T92C allele of the tetraspanin gene HaTSPAN1 confers dominant resistance to Bacillus thuringiensis toxin Cry1Ac

Transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) have revolutionized pest control. However, the evolution of resistance by target pests poses a significant threat to the long-term success of Bt crops. Understanding the genetics and mechanisms underlying Bt resistance is crucial for developing resistance detection methods and management tactics. The T92C mutation in a tetraspanin gene (HaTSPAN1), resulting in the L31S substitution, is associated with dominant resistance to Cry1Ac in a major pest, Helicoverpa armigera. Previous studies using CRISPR/Cas9 technique have demonstrated that knockin of the HaTSPAN1 T92C mutation confers a 125-fold resistance to Cry1Ac in the susceptible SCD strain of H. armigera. In this study, we employed the piggyBac transposon system to create two transgenic H. armigera strains based on SCD: one expressing the wild-type HaTSPAN1 gene (SCD-TSPANwt) and another expressing the T92C mutant form of HaTSPAN1 (SCD-TSPANmt). The SCD-TSPANmt strain exhibited an 82-fold resistance to Cry1Ac compared to the recipient SCD strain, while the SCD-TSPANwt strain remained susceptible. The Cry1Ac resistance followed an autosomal dominant inheritance mode and was genetically linked with the transgene locus in the SCD-TSPANmt strain of H. armigera. Our results further confirm the causal association between the T92C mutation of HaTSPAN1 and dominant resistance to Cry1Ac in H. armigera. Additionally, they suggest that the piggyBac-mediated transformation system we used in H. armigera is promising for functional investigations of candidate Bt resistance genes from other lepidopteran pests.

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来源期刊
CiteScore
7.00
自引率
8.50%
发文量
238
审稿时长
4.2 months
期刊介绍: Pesticide Biochemistry and Physiology publishes original scientific articles pertaining to the mode of action of plant protection agents such as insecticides, fungicides, herbicides, and similar compounds, including nonlethal pest control agents, biosynthesis of pheromones, hormones, and plant resistance agents. Manuscripts may include a biochemical, physiological, or molecular study for an understanding of comparative toxicology or selective toxicity of both target and nontarget organisms. Particular interest will be given to studies on the molecular biology of pest control, toxicology, and pesticide resistance. Research Areas Emphasized Include the Biochemistry and Physiology of: • Comparative toxicity • Mode of action • Pathophysiology • Plant growth regulators • Resistance • Other effects of pesticides on both parasites and hosts.
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