CRISPR/Cas9 mediated ryanodine receptor I4790M knockin confers unequal resistance to diamides in Plutella xylostella
CRISPR/Cas9 mediated ryanodine receptor I4790M knockin confers unequal resistance to diamides in Plutella xylostella
复制标题
CRISPR/Cas9介导的兰尼碱受体I4790M敲入赋予小菜蛾对二酰胺的不同抗性
DOI：
10.1016/j.ibmb.2020.103453
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发表时间：
2020
影响因子：
3.8
通讯作者：
Yidong Wu
中科院分区：
文献类型：
--
作者：
Xingliang Wang;Xiaowei Cao;Dong Jiang;Yihua Yang;Yidong Wu
小菜蛾（Plutella xylostella）是世界范围内芸苔属植物的主要害虫，对几乎所有的商业杀虫剂都产生了抗药性，包括最新的双酰胺类杀虫剂。一些研究表明，RyR基因的G4946 E和I4790 M突变与昆虫的二酰胺抗性密切相关。虽然G4946 E在赋予二酰胺抗性表型中的关键功能作用已通过不同物种中的几项研究证实，但没有直接证据明确证实单一I4790 M突变在二酰胺抗性中的功能意义。在此，我们成功地构建了一个敲入纯合菌株（I4790 M-KI）的P。使用CRISPR/Cas9偶联同源定向修复方法将I4790 M引入RyR。与背景敏感的IPP-S菌株相比，I4790 M-KI菌株对邻苯二甲酸二酰胺氟虫双酰胺表现出中等抗性（40.5倍），对邻氨基苯甲酰胺氯虫苯甲酰胺（6.0倍）和氰虫苯甲酰胺（7.7倍）表现出低抗性，对茚虫威和高效氯氰菊酯的毒性没有变化。此外，获得性氟虫双胺抗性以常染色体隐性模式遗传，并与I4790 M-KI菌株中RyR的I4790 M突变显著连锁。我们的研究结果为PxRyR的I4790 M突变与氟虫双胺中度耐药的因果关系提供了体内功能证据。小菜蛾，并支持的假设，二酰胺类具有不同的相互作用与RyRs。
The diamondback mothPlutella xylostellais a major destructive pest of Brassica worldwide.P. xylostellahas evolved resistance to nearly all commercial insecticides used for its control, including the most recent chemical class, diamide insecticides. Several studies show that the G4946E and I4790M mutations of ryanodine receptor (RyR) are strongly associated with diamide resistance in insects. While the pivotal functional role of G4946E in conferring diamide resistance phenotype has confirmed by several studies in different species, no direct evidence has unambiguously confirmed the functional significance of the single I4790M mutation in diamide resistance. Here, we successfully constructed a knockin homozygous strain (I4790M-KI) ofP. xylostellausing CRISPR/Cas9 coupled with homology directed repair approach to introduce I4790M into RyR. When compared with the background susceptible IPP-S strain, the manipulated I4790M-KI strain exhibited moderate resistance to the phthalic acid diamide flubendiamide (40.5-fold) and low resistance to anthranilic diamides chlorantraniliprole (6.0-fold) and cyantraniliprole (7.7-fold), with no changes to the toxicities of indoxacarb and β-cypermethrin. Furthermore, the acquired flubendiamide resistance was inherited in an autosomally recessive mode and significantly linked with the I4790M mutation of RyR in this I4790M-KI strain. Our findings providein vivofunctional evidence for the causality of I4790M mutation of PxRyR with moderate levels of resistance to flubendiamide inP. xylostella, and support the hypothesis that the diamide classes have different interactions with RyRs.