Journal of Integrative Agriculture ›› 2024, Vol. 23 ›› Issue (5): 1604-1617.DOI: 10.1016/j.jia.2023.11.001

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钙粘蛋白作为美州棉铃虫对Cry1Ac 和Cry2Ab交互或双重抗性共同机制的功能评估

  

  • 收稿日期:2023-05-16 接受日期:2023-08-24 出版日期:2024-05-20 发布日期:2024-04-23

Functional assessment of cadherin as a shared mechanism for cross/dual resistance to Cry1Ac and Cry2Ab in Helicoverpa zea

Jizhen Wei1, 2*, Min Zhang2, 3*, Pin Li1, Zhongyuan Deng3, Xinming Yin1, Shiheng An1#, Xianchun Li2#    

  1. 1 State Key Laboratory of Wheat and Maize Crop Science/College of Plant Protection, Henan Agricultural University, Zhengzhou 450046, China

    2 Department of Entomology and BIO5 Institute, University of Arizona, Tucson, AZ 85721, USA

    3 School of Agricultural Sciences, Zhengzhou University, Zhengzhou 450001, China

  • Received:2023-05-16 Accepted:2023-08-24 Online:2024-05-20 Published:2024-04-23
  • About author:Jizhen Wei, E-mail: weijizhen1986@163.com; Min Zhang, E-mail: 418078760@qq.com; #Correspondence Shiheng An, E-mail: anshiheng@aliyun.com; Xianchun Li, E-mail: lxc@email.arizona.edu * These authors contributed equally to this study.
  • Supported by:
    This research was funded by the USDA National Institute of Food and Agriculture (Hatch Grant ARZT-1360890-H31-164 and multi-state grant ARZT-1370680-R31-172 (NC246)), the National Natural Science Foundation of China (NSFC)–Henan Joint Major Grant (U2004206), the State Key Laboratory of Cotton Biology Open Fund, Zhengzhou University, China (CB2020A06), the Henan Agriculture Research System, China (HARS-22-09-G3), and the earmarked fund for China Agriculture Research System (CARS-27).

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摘要：

美洲棉铃虫（Helicoverpa zea）是转多个苏云金芽孢杆菌（Bacillus thuringiensis，Bt）杀虫蛋白（Cry1、Cry2和/或Vip3Aa）基因作物的主要害虫。实验室和田间都发现美洲棉铃虫对Cry1Ac/Cry2Ab的交互抗性或双重抗性。但是，普遍存在的美洲棉铃虫对Cry1Ac/Cry2Ab的交互抗性或双重抗性是共同作用机制还是两个独立的抗性机制引起的还不清楚。之前报道钙黏蛋白是Cry1Ac的确定受体，有研究也表明钙黏蛋白至少在三种鳞翅目中是Cry2Ab的受体。为了验证钙黏蛋白是否作为Cry1Ac/Cry2Ab交互抗性或双重抗性的共同作用机制，我们克隆了美洲棉铃虫钙黏蛋白基因并通过过表达和干扰该基因的方式分别探究了其在Cry1Ac和Cry2Ab作用机制中的功能。在美洲棉铃虫中肠细胞，脂肪体细胞及Sf9细胞中表达美洲棉铃虫钙黏蛋白基因导致三种细胞对活化的Cry1Ac敏感性增强，但对Cry2Ab的敏感性不变；对应的在美洲棉铃虫中肠细胞和脂肪体细胞中干扰钙黏蛋白基因导致细胞对Cry1Ac敏感性降低，但对Cry2Ab的敏感性不变。同样，用钙黏蛋白siRNA在活体中抑制钙黏蛋白的表达增加了幼虫对Cry1Ac的抗性。这些结果清楚地表明，HzCadheirn不是Cry2Ab的受体，因此不可能是美洲棉铃虫对Cry1Ac和Cry2Ab的交互抗性和双重抗性的共同机制。

Abstract:

Helicoverpa zea is a major target pest of pyramided transgenic crops expressing Cry1, Cry2 and/or Vip3Aa proteins from Bacillus thuringiensis (Bt) in the United States.  Laboratory-selected Cry1Ac/Cry2Ab cross resistance and field-evolved practical dual resistance of H. zea to these two toxins have been widely reported.  Whether the widespread Cry1Ac/Cy2Ab dual resistance of H. zea has resulted from the selection of one shared or two independent resistance mechanisms by pyramided Bt crops remains unclear.  Cadherin is a well-confirmed receptor of Cry1Ac and a suggested receptor of Cry2Ab in at least three Lepidopteran species.  To test whether cadherin may serve as one shared mechanism for the cross and dual resistance of H. zea to Cry1Ac and Cry2Ab, we cloned H. zea cadherin (HzCadherin) cDNA and studied its functional roles in the mode of action of Cry1Ac and Cry2Ab by gain- and loss-of-function analyses.  Heterologous expression of HzCadherin in H. zea midgut, H. zea fat body and Sf9 cells made all three of these cell lines more susceptible to activated Cry1Ac but not activated Cry2Ab, whereas silencing HzCadherin of H. zea midgut and fat body cells significantly reduced the susceptibility to Cry1Ac but not Cry2Ab.  Likewise, suppressing HzCadherin with siRNA made H. zea larvae resistant to Cry1Ac.  These results clearly demonstrate that HzCadherin is not a receptor for Cry2Ab, and thus it is unlikely to serve as one shared mechanism for the cross and dual resistance of H. zea to Cry1Ac and Cry2Ab.

Key words: Bt crops ,  cadherin ,  Cry toxin ,  receptor ,  resistance mechanism