Meng Xiangkun, Li Chunrui, Bao Haibo, Fang Jichao, Liu Zewen, Zhang Yixi
Key Laboratory of Integrated Management of Crop Diseases and Pests (Ministry of Education), College of Plant Protection, Nanjing Agricultural University, Weigang 1, Nanjing 210095, China.
Key Laboratory of Integrated Management of Crop Diseases and Pests (Ministry of Education), College of Plant Protection, Nanjing Agricultural University, Weigang 1, Nanjing 210095, China; Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, St. Zhongling 50, Nanjing 210014, China.
Pestic Biochem Physiol. 2015 Nov;125:26-30. doi: 10.1016/j.pestbp.2015.06.006. Epub 2015 Jun 20.
The pond wolf spider (Pardosa pseudoannulata) is an important predatory enemy against several insect pests and showed relative different sensitivities to organophosphate and carbamate insecticides compared to insect pests. In our previous studies, two acetylcholinesterases were identified in P. pseudoannulata and played important roles in insecticide sensitivities. In order to understand the contributions of the two acetylcholinesterases to insecticide sensitivities, we firstly employed the RNAi technology in the spider. For a suitable microinjection RNAi method, the injection site, injection volume and interference time were optimized, which then demonstrated that the injection RNAi method was applicable in this spider. With the new RNAi method, it was revealed that both Pp-AChE1 and Pp-AChE2, encoded by genes Ppace1 and Ppace2, were the targets of organophosphate insecticides, but Pp-AChE1 would be more important. In contrast, the carbamate acted selectively on Pp-AChE1. The results showed that Pp-AChE1 was the major catalytic enzyme in P. pseudoannulata and the major target of organophosphate and carbamate insecticides. In a word, an RNAi method was established in the pond wolf spider, which further validated the importance of two acetylcholinesterases in insecticide sensitivities in this spider.
拟环纹豹蛛（Pardosa pseudoannulata）是多种害虫的重要捕食性天敌，与害虫相比，其对有机磷和氨基甲酸酯类杀虫剂表现出相对不同的敏感性。在我们之前的研究中，在拟环纹豹蛛中鉴定出两种乙酰胆碱酯酶，它们在杀虫剂敏感性方面发挥着重要作用。为了了解这两种乙酰胆碱酯酶对杀虫剂敏感性的贡献，我们首先在蜘蛛中采用了RNA干扰技术。对于合适的显微注射RNA干扰方法，对注射部位、注射体积和干扰时间进行了优化，结果表明注射RNA干扰方法适用于这种蜘蛛。利用新的RNA干扰方法发现，由Ppace1和Ppace2基因编码的Pp-AChE1和Pp-AChE2都是有机磷杀虫剂的作用靶点，但Pp-AChE1更为重要。相比之下，氨基甲酸酯类选择性作用于Pp-AChE1。结果表明，Pp-AChE1是拟环纹豹蛛中的主要催化酶，也是有机磷和氨基甲酸酯类杀虫剂的主要作用靶点。总之，在拟环纹豹蛛中建立了一种RNA干扰方法，进一步验证了两种乙酰胆碱酯酶在该蜘蛛杀虫剂敏感性中的重要性。
