Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (8): 3127-3140.DOI: 10.1016/j.jia.2024.09.022

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鳞翅目害虫四次跨膜蛋白TspC5的物种特异性进化与Bt毒素Cry1Ac显性抗性

  

  • 收稿日期:2024-08-02 修回日期:2024-09-26 接受日期:2024-09-10 出版日期:2025-07-20 发布日期:2025-07-17

Species-specific evolution of lepidopteran TspC5 tetraspanins associated with dominant resistance to Bacillus thuringiensis toxin Cry1Ac

Chenyang Wang, Yinuo Zhang, Qiming Sun, Lin Li, Fang Guan, Yazhou He, Yidong Wu#   

  1. College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2024-08-02 Revised:2024-09-26 Accepted:2024-09-10 Online:2025-07-20 Published:2025-07-17
  • About author:Chenyang Wang, E-mail: cywang@stu.njau.edu.cn; #Correspondence Yidong Wu, E-mail: wyd@njau.edu.cn
  • Supported by:

    This research was primarily funded by a grant from the National Natural Science Foundation of China (31930093).  Additional support was provided by the Natural Science Foundation of Jiangsu Province, China (BK20230983), and the Project of Fund for Stable Support to Agricultural Sci-Tech Renovation, China (xjnkywdzc-2022004).

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摘要：

Bt蛋白是苏云金芽孢杆菌Bacillus thuringiensis（Bt）在生长阶段产生的杀虫蛋白，已被广泛应用于转基因作物中，有效控制了多种重要的鳞翅目和鞘翅目害虫。然而，这些害虫对Bt蛋白的抗性进化对转基因作物的长期可持续性构成了挑战。棉铃虫Helicoverpa armigera是一种重要的鳞翅目农业害虫。之前的研究发现棉铃虫一个四次跨膜蛋白基因HarmTspC5（HaTSPAN1）中的L31S突变赋予了棉铃虫对Bt蛋白Cry1Ac的显性抗性。然而，L31S突变在其他鳞翅目物种中的作用尚不明确。本研究的分析表明，鳞翅目昆虫TspC5基因的进化具有物种特异性。为探究鳞翅目昆虫TspC5基因在赋予Cry1Ac显性抗性中的作用，本研究基于piggyBac遗传转化系统构建了四个转基因棉铃虫品系。这些品系表达了携带L31S同源突变的TspC5蛋白，它们分别来自与棉铃虫近缘的美洲棉铃虫Helicoverpa zea、烟青虫Helicoverpa assulta和烟芽夜蛾Heliothis virescens，以及相对远缘的小菜蛾Plutella xylostella。生物测定显示，与背景品系SCD相比，表达HzeaTspC5-L31S、HassTspC5-L31S或HvirTspC5-L31S的转基因品系对Cry1Ac表现出显著抗性，抗性倍数分别为10.0倍、21.4倍和81.1倍，而表达PxylTspC5-L27S的品系对Cry1Ac仍然敏感。此外，三个转基因品系的Cry1Ac抗性表型以常染色体显性的方式遗传，并与外源TspC5突变体紧密连锁。这些发现揭示了棉铃虫近缘物种的TspC5基因在介导Cry1Ac显性抗性中的保守作用，并为评估与四次跨膜蛋白突变相关的抗性风险和制定适应性抗性治理策略提供了重要依据。

Abstract:

Transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) have proven to be highly effective in managing some key pests.  However, the evolution of resistance by the target pests threatens the sustainability of Bt crops.  The L31S mutation in a tetraspanin encoded by HarmTspC5 (previously known as HaTSPAN1) has been shown to confer dominant resistance to the Bt protein Cry1Ac in Helicoverpa armigera, a globally damaging lepidopteran pest.  However, the broader implications of the L31S mutation in the tetraspanins of other lepidopteran species remain unclear.  The evolutionary analyses in this study indicate that TspC5s have evolved in a species-specific manner among the lepidopteran insects.  To investigate the role of TspC5s in conferring dominant resistance to Cry1Ac, we used the piggyBac-based transformation system to generate four transgenic H. armigera strains that express exogenous TspC5 variants from three phylogenetically close species (Helicoverpa zea, Helicoverpa assulta and Heliothis virescens) and one phylogenetically distant species (Plutella xylostella).  In comparison with the background SCD strain of H. armigera, the transgenic strains expressing HzeaTspC5-L31S, HassTspC5-L31S, or HvirTspC5-L31S exhibited significant resistance to Cry1Ac (10.0-, 21.4-, and 81.1-fold, respectively), whereas the strain expressing PxylTspC5-L27S remained susceptible.  Furthermore, the Cry1Ac resistant phenotypes followed an autosomal dominant inheritance pattern and were closely linked to the introduced mutant TspC5s.  These findings reveal the conserved role of TspC5s from Helicoverpa and Heliothis species in mediating the dominant resistance to Cry1Ac, and they provide crucial insights for assessing resistance risks related to mutant tetraspanins and devising adaptive resistance management strategies for these major lepidopteran pests.


Key words: tetraspanin ,  Bt resistance ,  Cry1Ac ,  piggyBac system ,  Helicoverpa armigera