沈宁,黄宇星,唐涛,彭英传,张莹,赵春青,2023,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[J].环境昆虫学报,45(1):269-276
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 dsECR of Henosepilachna vigintioctopunctata
  
DOI:
中文关键词:  纳米材料  层状双氢氧化物  LDH  茄二十八星瓢虫  RNA干扰
英文关键词:Nanomaterials  layered double hydroxide  LDH  Henosepilachna vigintioctopunctata  RNA interference
基金项目: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)
作者单位
沈宁,黄宇星,唐涛,彭英传,张莹,赵春青 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 
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中文摘要:
      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。
英文摘要:
      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 dsEcR were assembled to form a complex of LDH-dsEcR. 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 dsECR 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-dsEcR, 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.
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