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通过小RNA深度测序鉴定用于控制甜菜夜蛾的合成微小RNA模拟物的微小RNA。

Identification of microRNAs by small RNA deep sequencing for synthetic microRNA mimics to control Spodoptera exigua.

作者信息

Zhang Yu Liang, Huang Qi Xing, Yin Guo Hua, Lee Samantha, Jia Rui Zong, Liu Zhi Xin, Yu Nai Tong, Pennerman Kayla K, Chen Xin, Guo An Ping

机构信息

Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China.

Department of Plant Biology and Pathology, Rutgers, The State University of New Jersey, New Brunswick, NJ 08901, USA.

出版信息

Gene. 2015 Feb 25;557(2):215-21. doi: 10.1016/j.gene.2014.12.038. Epub 2014 Dec 17.

Abstract

Beet armyworm, Spodoptera exigua, is a major pest of cotton around the world. With the increase of resistance to Bacillus thuringiensis (Bt) toxin in transgenic cotton plants, there is a need to develop an alternative control approach that can be used in combination with Bt transgenic crops as part of resistance management strategies. MicroRNAs (miRNAs), a non-coding small RNA family (18-25 nt), play crucial roles in various biological processes and over-expression of miRNAs has been shown to interfere with the normal development of insects. In this study, we identified 127 conserved miRNAs in S. exigua by using small RNA deep sequencing technology. From this, we tested the effects of 11 miRNAs on larval development. We found three miRNAs, Sex-miR-10-1a, Sex-miR-4924, and Sex-miR-9, to be differentially expressed during larval stages of S. exigua. Oral feeding experiments using synthetic miRNA mimics of Sex-miR-10-1a, Sex-miR-4924, and Sex-miR-9 resulted in suppressed growth of S. exigua and mortality. Over-expression of Sex-miR-4924 caused a significant reduction in the expression level of chitinase 1 and caused abortive molting in the insects. Therefore, we demonstrated a novel approach of using miRNA mimics to control S. exigua development.

摘要

甜菜夜蛾(Spodoptera exigua)是全球棉花的主要害虫。随着转基因棉花植株对苏云金芽孢杆菌(Bt)毒素的抗性增加,需要开发一种替代控制方法,作为抗性管理策略的一部分与Bt转基因作物联合使用。微小RNA(miRNA)是一种非编码小RNA家族(18 - 25个核苷酸),在各种生物学过程中发挥关键作用,并且已证明miRNA的过表达会干扰昆虫的正常发育。在本研究中,我们利用小RNA深度测序技术在甜菜夜蛾中鉴定出127个保守的miRNA。据此,我们测试了11个miRNA对幼虫发育的影响。我们发现三个miRNA,即Sex-miR-10-1a、Sex-miR-4924和Sex-miR-9,在甜菜夜蛾幼虫阶段差异表达。使用Sex-miR-10-1a、Sex-miR-4924和Sex-miR-9的合成miRNA模拟物进行口服喂养实验,导致甜菜夜蛾生长受抑制和死亡。Sex-miR-4924的过表达导致几丁质酶1的表达水平显著降低,并导致昆虫蜕皮失败。因此,我们证明了一种使用miRNA模拟物控制甜菜夜蛾发育的新方法。

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