文献类型：期刊文献

中文题名：基于滚环转录技术的小菜蛾RNAi靶标致死基因筛选及活性评价

英文题名：Screening and activity evaluation of lethal RNAi target genes in Plutella xylostella based on RCT technology

作者：须志平[1];王新骞[1];刘晓颖[1];刘彬[2];张阳[1]

机构：[1]华东理工大学药学院,上海市化学生物学(芳香杂环)重点实验室,上海200237;[2]上海市青浦区农业技术推广服务中心,上海201700

年份：2025

卷号：27

期号：5

起止页码：849

中文期刊名：农药学学报

外文期刊名：Chinese Journal of Pesticide Science

收录：;EI(收录号：20260319913445);北大核心:【北大核心2023】；

基金：上海市农业科技创新项目(No.K2025008)。

语种：中文

中文关键词：小菜蛾;RNA干扰;滚环转录;RNA生物农药;海藻糖酶;V-ATP酶;RPS13

外文关键词：Plutella xylostella;RNAi;rolling circle transcription;RNA biopesticide;trehalase;V-ATPase;RPS13

摘要：为筛选可防控小菜蛾(Plutella xylostella)的关键RNA干扰(RNAi)靶标基因,针对性选取海藻糖酶(trehalase)基因(PxTRE)、V-ATP酶E亚基(vacuolar ATP synthethase subunit E)基因(PxATPE)、RPS13基因(PxRPS13)和几丁质酶(chitinase)基因(PxCHT),采用滚环转录(RCT)技术设计并合成了多种RNA微球(RMS),采用饲喂法处理后,观察并测定了所得RNA微球对小菜蛾3龄幼虫生长发育的影响和致死效果。结果显示:处理48 h后,靶标基因沉默效果较为显著,RMS-PxTRE、RMS-PxATPE、RMS-PxRPS13和RMS-PxCHT处理组幼虫的mRNA基因表达水平相较于RMS-EGFP对照组分别降低了74.84%、73.36%、62.90%和38.10%(P<0.05)。此外,靶向双基因的多组分RMS-PxTA(RMS-PxTRE/ATPE)和RMS-PxTC(RMS-PxTRE/CHT)能同时显著降低PxTRE、PxATPE和PxCHT基因的表达水平;RMSPxTA处理组中,PxTRE和PxATPE基因表达量较对照组分别降低了86.16%和96.38%(P<0.05);RMS-PxTC处理组中,PxTRE和PxCHT基因表达量分别比对照组降低了45.22%和28.02%(P<0.05)。处理72 h后,RMS-PxTRE、RMS-PxATPE、RMS-PxCHT、RMS-PxRPS13、RMS-PxTA和RMS-PxTC处理组幼虫的死亡率分别为62.22%、86.67%、60.00%、57.78%、35.56%和66.67%;且处理组的存活幼虫表现出取食减少、体长缩短以及蜕皮失败等现象。研究结果表明,RCT技术可用于小菜蛾RNAi靶标基因的快速筛选,PxATPE基因可作为小菜蛾高效致死的RNA生物农药潜在靶标基因,推测其在小菜蛾RNAi绿色防控方面可能具有较好的应用前景。
To screen for key RNA interference(RNAi)target genes for controlling Plutella xylostella,trehalase(PxTRE),vacuolar ATP synthase subunit E(PxATPE),ribosomal protein S13(PxRPS13),and chitinase(PxCHT)were selected.Multiple RNA microspheres(RMS)were designed and synthesized using rolling circle transcription(RCT)technology.After administering the RMS to third-instar larvae via oral feeding,larval development and mortality were assessed.The results showed that gene silencing was significant after 48 hours of treatment.Compared to the RMS-EGFP control group,mRNA expression levels of PxTRE,PxATPE,PxRPS13,and PxCHT were reduced by 74.84%,73.36%,62.90%,and 38.10%,respectively(P<0.05).In addition,multi-RMS RMS-PxTA(RMS-PxTRE/ATPE)and RMS-PxTC(RMS-PxTRE/CHT)significantly downregulated the expression of the corresponding genes.In the RMS-PxTA treatment,the expression levels of PxTRE and PxATPE were reduced by 86.16%and 96.38%,respectively(P<0.05).Similarly,in the RMS-PxTC treatment,the expression levels of PxCHT were decreased by 45.22%and 28.02%,respectively(P<0.05).After 72 hours,the larval mortality rates in the RMS-PxTRE、RMS-PxATPE、RMS-PxCHT和RMS-PxRPS13、RMS-PxTA and RMS-PxTC treatments were 62.22%,86.67%,60.00%,57.78%,35.56%,and 66.67%,respectively.Surviving larvae in the treatment groups exhibited reduced feeding,shortened body length,and molting failure.These findings suggest that RCT technology is a viable method for rapid screening of RNAi target genes in P.xylostella.Among the tested target genes,PxATPE demonstrates potential as a highly effective target for RNA-based biopesticide,offering promising prospects for environmentally friendly control of P.xylostella.