Knockout of three aminopeptidase N genes does not affect susceptibility of Helicoverpa armigera larvae to Bacillus thuringiensis Cry1A and Cry2A toxins
Knockout of three aminopeptidase N genes does not affect susceptibility of Helicoverpa armigera larvae to Bacillus thuringiensis Cry1A and Cry2A toxins
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敲除三个氨肽酶 N 基因不影响棉铃虫幼虫对苏云金芽孢杆菌 Cry1A 和 Cry2A 毒素的敏感性
DOI：
10.1111/1744-7917.12666
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发表时间：
2019-03
期刊：
影响因子：
4
通讯作者：
Wu Yi-Dong
中科院分区：
文献类型：
--
作者：
Wang Jing;Zuo Ya-Yun;Li Ling-Li;Wang Hui;Liu Shao-Yan;Yang Yi-Hua;Wu Yi-Dong
苏云金芽孢杆菌（Bt）杀虫毒素已在全球范围内通过喷洒或转基因作物用于控制农业昆虫。Bt毒素与靶标昆虫中肠上皮细胞上的特异性受体结合是其作用方式的关键步骤。以往的研究表明，氨肽酶N1（APN 1）作为一个受体或推定的受体在几个鳞翅目昆虫包括棉铃虫通过证据RNA干涉为基础的基因沉默方法。在目前的研究中，我们使用成簇规则间隔回文重复序列（CRISPR）/CRISPR相关蛋白9介导的基因敲除来测试APN在Bt毒素作用模式中的作用。在H. armigera以建立三个纯合敲除菌株。体外定性结合研究表明，APN敲除对Cry 1Ac和Cry 2Ab与中肠刷状缘膜囊泡的结合没有明显影响。生物测定结果表明，与SCD株相比，三种基因敲除株对Cry 1A或Cry 2A毒素的敏感性均无显著变化。这表明，我们测试的三个HaAPN基因可能不是关键的Cry 1A或Cry 2A毒素在H. armigera。
Bacillus thuringiensis (Bt) insecticidal toxins have been globally utilized for control of agricultural insects through spraying or transgenic crops. Binding of Bt toxins to special receptors on midgut epithelial cells of target insects is a key step in the mode of action. Previous studies suggested aminopeptidase N1 (APN1) as a receptor or putative receptor in several lepidopteran insects including Helicoverpa armigera through evidence from RNA interefence‐based gene silencing approaches. In the current study we tested the role of APNs in the mode of action of Bt toxins using clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR‐associated protein 9‐mediated gene knockout. Three APN genes (HaAPN1, HaAPN2 and HaAPN5) were individually knocked out in a susceptible strain (SCD) of H. armigera to establish three homozygous knockout strains. Qualitative in vitro binding studies indicated binding of Cry1Ac or Cry2Ab to midgut brush border membrane vesicles was not obviously affected by APN knockout. Bioassay results showed that none of the three knockouts had significant changes in susceptibility to Cry1A or Cry2A toxins when compared with the SCD strain. This suggests that the three HaAPN genes we tested may not be critical in the mode of action of Cry1A or Cry2A toxins in H. armigera.
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影响因子：
3.8
作者：
Shaoping Zhang;Hongmei Cheng;Yulin Gao;Gui-rong Wang;G. Liang;Kongming Wu
通讯作者：
Shaoping Zhang;Hongmei Cheng;Yulin Gao;Gui-rong Wang;G. Liang;Kongming Wu
影响因子：
3.7
作者：
Zhang H;Yin W;Zhao J;Jin L;Yang Y;Wu S;Tabashnik BE;Wu Y
通讯作者：
Wu Y
影响因子：
4.5
作者：
Gahan LJ;Pauchet Y;Vogel H;Heckel DG
通讯作者：
Heckel DG
影响因子：
3.7
作者：
Tay WT;Soria MF;Walsh T;Thomazoni D;Silvie P;Behere GT;Anderson C;Downes S
通讯作者：
Downes S
影响因子：
3.8
作者：
Ping Wang;Xin Zhang;Jie Zhang
通讯作者：
Ping Wang;Xin Zhang;Jie Zhang