纳米材料层状双氢氧化物(LDH)对茄二十八星瓢虫dsECR致死效应的影响  

Lethal effects of nanomaterial layered double hydroxide (LDH)to ds ECR of Henosepilachna vigintioctopunctata

作　　者：沈宁 黄宇星 唐涛[3] 彭英传 张莹 赵春青[1] SHEN Ning;HUANG Yu-Xing;TANG Tao;PENG Ying-Chuan;ZHANG Ying;ZHAO Chun-Qing(College of Plant Protection,Nanjing Agricultural University,Nanjing 210095,China;School of Material Science and Engineering,Nanchang University,Nanchang 330031,China;Institute of Plant Protection,Hunan Academy of Agricultural Sciences,Changsha 410125,China)

机构地区：[1]南京农业大学植物保护学院,南京210095 [2]南昌大学材料科学与工程学院,南昌330031 [3]湖南省农业科学院植物保护研究所,长沙410125

出　　处：《环境昆虫学报》2023年第1期269-276,共8页Journal of Environmental Entomology

基　　金：国家重点研发计划(2017YFD0200900);国家自然科学基金(31871995);江苏省研究生科研与实践创新计划(SJCX20_0211)。

摘　　要：纳米材料被视为重要的RNA干扰增效物质。层状双氢氧化物(LDH)是具有生物相容性的可降解纳米材料,可以显著提高dsRNA防控黄瓜花叶病毒的效果。为明确LDH能否提高昆虫RNA干扰的效果,以期促进RNA生物农药在农业害虫防治中的应用和推广,本研究选择茄二十八星瓢虫Henosepilachna vigintioctopunctata的EcR基因进行RNA干扰介导的致死效应研究。将LDH与dsEcR组装形成复合物LDH-dsEcR,利用琼脂糖凝胶电泳确定LDH-dsEcR的最佳装载比例。通过静置法检测LDH-dsEcR的稳定性,借助透射电子显微镜验证其组装状态。采用浸叶法测定LDH-dsEcR的增效性,使用体视镜观察其对试虫形态的影响。结果表明:LDH与dsEcR可形成稳定的复合物,其最佳装载比为16∶1。LDH对茄二十八星瓢虫无毒性,但经LDH与dsEcR处理,部分试虫虫体呈黑褐色,或无法蜕皮化蛹,或维持在预蛹状态。遗憾的是LDH-dsEcR复合物的增效作用不明显,推测昆虫肠道的酸碱度可能是影响LDH-dsRNA增效的因素之一。本研究首次为利用LDH研究昆虫RNA生物农药提供了可观的理论基础。Nanomaterial is considered as an important RNA interference(RNAi)synergist.Layered double hydroxide(LDH)is a biocompatible and biodegradable nanomaterial and plays a significant synergism in control of tobacco cucumber mosaic virus using double-stranded RNA(dsRNA).In this study,we aimed to determine whether LDH can enhance the effect of insect RNAi,so as to promote the application of RNA biopesticide in control of agricultural insects.The EcR gene of Henosepilachna vigintioctopunctata was selected to study its synergism on RNAi,and the LDH and ds EcR were assembled to form a complex of LDH-ds EcR.The optimal loading ratio of the complex was determined by the gel electrophoresis,the stability of the complex was detected by the static method,and the assembly status of the complex was verified by the transmission electron microscope.The synergism of the complex was determined by the leaf-dip method,and the effect of the complex on the morphology of H.vigintioctopunctata was observed by the stereoscope.LDH and ds ECR can form a stable complex with the optimum loading ratio of 16∶1.LDH was not toxic to H.vigintioctopunctata,but the complex was lethal.After treated with the LDH-ds EcR,some H.vigintioctopunctata larvae become dark brown,or cannot molt into pupation or stay at the stage of prepupation.Regretfully,the synergism of the complex was not significant.We speculated that the pH of insect intestine might be one factor affecting the synergism of LDH-dsRNA complex.However,our result firstly provides the preliminary data for using LDH in biopesticide with dsRNA for insect control.

关 键 词：纳米材料 层状双氢氧化物 LDH 茄二十八星瓢虫 RNA干扰 

分 类 号：Q968.1[生物学—昆虫学] S433[农业科学—农业昆虫与害虫防治]