纳米载体介导的乙酰胆碱酯酶RNA干扰对朱砂叶螨生长发育的影响  

Effects of nanocarrier-mediated RNA interference with the acetylcholinesterase gene on the growth and development of Tetranychus cinnabarinus

作　　者：王谦稳 卜春亚[1,2] 陈耔霖 马雪晶 任争光[1,2] WANG Qianwen;BU Chunya;CHEN Zilin;MA Xuejing;REN Zhengguang(College of Biological and Resource Environment,Beijing University of Agriculture,Beijing 102206,China;Key Laboratory for Northern Urban Agriculture of Ministry of Agriculture and Rural Affairs,Beijing University of Agriculture,Beijing 102206,China)

机构地区：[1]北京农学院生物与资源环境学院,北京102206 [2]北京农学院农业农村部华北都市农业重点实验室,北京102206

出　　处：《应用与环境生物学报》2024年第5期975-980,共6页Chinese Journal of Applied and Environmental Biology

基　　金：北京市自然科学基金项目(6212004);国家自然科学基金项目(31670648)资助。

摘　　要：为探明纳米载体介导的RNA干扰乙酰胆碱酯酶(ace)基因表达对朱砂叶螨的影响,通过NCBI上获取朱砂叶螨的ace 1基因,利用RT-qPCR技术对ace 1基因在朱砂叶螨各个时期的表达水平进行检测,并利用纳米递呈dsRNA-ace 1的RNAi技术,检测其对朱砂叶螨的影响.结果显示,乙酰胆碱酯酶基因在朱砂叶螨4个时期均有表达,其中以成螨期表达量最高,其次是卵、若螨和幼螨.通过叶碟饲喂dsRNA-ace 1能够有效抑制朱砂叶螨ace 1的表达,ace 1基因表达水平下降57%,72 h致死率达到41.77%,同时对朱砂叶螨的产卵、孵育造成显著的影响.通过纳米递呈dsRNA-ace 1能够显著提高对乙酰胆碱酯酶基因表达的抑制,抑制率同比未纳米递呈dsRNA-ace 1提高24%,致死率同比增幅32.08%,产卵率与孵化率也均有显著降低.同时进行微滴法施用dsRNA-ace 1/SPc复合物,对朱砂叶螨ace 1的RNA干扰效果,和叶碟饲喂法差异不显著.本研究表明纳米载体递呈dsRNA-ace 1能够提高对ace 1基因表达的抑制,提高对朱砂叶螨的致死效果,结果可为后期以纳米载体介导的RNAi技术对害虫开展生物防治提供理论基础.(图7参27)This study was conducted to investigate the effect of nanocarrier-mediated RNA interference on acetylcholinesterase(ACE)gene expression in Tetranychus cinnabarinus.We obtained ace 1 from T.cinnabarinus and detected its gene expression level during each developmental stage of T.cinnabarinus using RT-qPCR.The lethal effect of nanodelivered dsRNA-ace 1 on T.cinnabarinus was detected using RNA interference(RNAi)technology.The results showed that AChE was expressed in all four developmental stages of T.cinnabarinus,with the highest expression in the adult mite stage,followed by the eggs,nymphs,and larvae.Feeding of double-stranded RNA(dsRNA)-ace 1 through leaf discs significantly inhibited ace 1 expression in T.cinnabarinus,with a 57% reduction in gene expression at 48 h and lethality of 41.77% at 72 h,while inhibiting egg laying and hatching of T.cinnabarinus.Nano-delivery of dsRNA-ace 1 efficiently inhibited expression of the acetylcholinesterase gene;the inhibition rate increased by 24%,lethality increased by 32.08% compared with that induced by dsRNA-ace 1 alone,and the egg laying and hatching rates were significantly reduced.There was no significant difference in the RNAi effects of microdroplet application of the dsRNA-ace 1/SPc complex compared and the leaf disc feeding method.These results indicate that nanocarrier delivery of dsRNA-ace 1 can increase the inhibition efficiency of ace 1 expression and improve its lethal effect on T.cinnabarinus.These results provide a foundation for biological control of pests using nanocarrier-mediated RNAi technology.

关 键 词：朱砂叶螨 乙酰胆碱酯酶 基因克隆 纳米载体 RNA干扰 

分 类 号：S433.7[农业科学—农业昆虫与害虫防治]