Knocking Out of UDP–Glycosyltransferase Gene UGT2B10 via CRISPR/Cas9 in Helicoverpa armigera Reveals Its Function in Detoxification of Insecticides,Journal of Agricultural and Food Chemistry

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Knocking Out of UDP–Glycosyltransferase Gene UGT2B10 via CRISPR/Cas9 in Helicoverpa armigera Reveals Its Function in Detoxification of Insecticides
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2024-09-13 , DOI: 10.1021/acs.jafc.4c05055
Junyue Zheng ₁ , Xiangguang Chen ₂ , Yao Xie ₁ , Yu Zhang ₁ , Yun Huang ₁ , Peizhuo Wu ₁ , Jingshu Lv ₁ , Lihong Qiu ₁

Affiliation

The role of insect UDP-glycosyltransferases (UGTs) in the detoxification of insecticides has rarely been reported. A UGT gene UGT2B10 was previously found overexpressed in a fenvalerate-resistant strain of Helicoverpa armigera. Herein, UGT2B10 was cloned, and its involvement in insecticide detoxification was investigated. UGT2B10 was highly expressed in the larvae, mainly in the fat body and midgut. Treatment with UGT inhibitors 5-nitrouracil and sulfinpyrazone significantly enhanced the fenvalerate toxicity. Knocking down UGT2B10 by RNAi significantly increased the larvae mortality by 17.89%. UGT2B10 was further knocked out by CRISPR/Cas9, and a homozygous strain (HD-dUGT2B10) with a C-base deletion at exon 2 was obtained. The sensitivity of HD-dUGT2B10 to fenvalerate, deltamethrin, cyantraniliprole, acetamiprid, and lufenuron increased significantly, with sensitivity index increased 2.523-, 2.544-, 2.250-, 2.473-, and 3.556-fold, respectively. These results suggested that UGT2B10 was involved in the detoxification of H. armigera to insecticides mentioned above, shedding light upon further understanding of the detoxification mechanisms of insecticides by insect UGTs.

中文翻译：

通过 CRISPR/Cas9 敲除棉铃虫中的 UDP-糖基转移酶基因 UGT2B10，揭示其在杀虫剂解毒中的功能

昆虫UDP-糖基转移酶（UGT）在杀虫剂解毒中的作用鲜有报道。先前发现 UGT 基因UGT2B10在抗氰戊菊酯菌株中过度表达。在此，克隆了UGT2B10 ，并研究了其在杀虫剂解毒中的作用。 UGT2B10在幼虫中高表达，主要在脂肪体和中肠中。 UGT抑制剂5-硝基尿嘧啶和磺吡酮治疗显着增强了氰戊菊酯的毒性。通过RNAi敲除UGT2B10使幼虫死亡率显着增加17.89%。通过CRISPR/Cas9进一步敲除UGT2B10 ，获得2号外显子C碱基缺失的纯合菌株（HD-d UGT2B10 ）。 HD-d UGT2B10对氰戊菊酯、溴氰菊酯、氰虫苯甲酰胺、啶虫脒和虱螨脲的敏感性显着增加，敏感性指数分别增加2.523、2.544、2.250、2.473和3.556倍。这些结果表明UGT2B10参与了棉铃虫对上述杀虫剂的解毒，为进一步了解昆虫UGT对杀虫剂的解毒机制提供了线索。

更新日期：2024-09-13

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