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影响稻纵卷叶螟RNA干扰效率的双链RNA降解核酸酶的鉴定与特性分析

Identification and Characterization of a Double-Stranded RNA Degrading Nuclease Influencing RNAi Efficiency in the Rice Leaf Folder .

作者信息

Li Jiajing, Du Juan, Li Shangwei, Wang Xin

机构信息

Guizhou Provincial Key Laboratory for Agricultural Pest Management of Mountainous Regions, Institute of Entomology, Guizhou University, Guiyang 550025, China.

出版信息

Int J Mol Sci. 2022 Apr 2;23(7):3961. doi: 10.3390/ijms23073961.

DOI:10.3390/ijms23073961
PMID:35409320
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8999895/
Abstract

Rice leaf folder is one of the most serious pests of rice in rice-planting regions worldwide. DsRNA-degrading nucleases (dsRNases) are important factors in reducing the efficiency of RNA interference (RNAi) in different insects. In this study, a gene from () was cloned and characterized. The cDNA was 1395 bp in length, encoding 464 amino acids. The CmdsRNase zymoprotein contains a signal peptide and an endonuclease NS domain that comprises six active sites, three substrate-binding sites, and one Mg-binding site. The mature CmdsRNase forms a homodimer with a total of 16 α-helices and 20 β-pleated sheets. Homology and phylogenetic analyses revealed that CmdsRNase is closely related to dsRNase2 in . Expression pattern analysis by droplet digital PCR indicated that the expression levels of varied throughout the developmental stages of and in different adult tissues, with the highest expression levels in the fourth-instar larvae and the hemolymph. CmdsRNase can degrade dsRNA to reduce the efficiency of RNAi in . Co-silencing of ( from ) and CmdsRNase affected significantly the growth and development of and thus improved RNAi efficacy, which increased by 27.17%. These findings will be helpful for green control of and other lepidopteran pests by RNAi.

摘要

稻纵卷叶螟是全球水稻种植区最严重的水稻害虫之一。双链RNA降解核酸酶（dsRNases）是降低不同昆虫中RNA干扰（RNAi）效率的重要因素。在本研究中，克隆并鉴定了来自（）的一个基因。该cDNA长度为1395 bp，编码464个氨基酸。CmdsRNase酶蛋白包含一个信号肽和一个核酸内切酶NS结构域，该结构域包含六个活性位点、三个底物结合位点和一个镁结合位点。成熟的CmdsRNase形成一个同二聚体，共有16个α螺旋和20个β折叠片层。同源性和系统发育分析表明，CmdsRNase与（）中的dsRNase2密切相关。通过液滴数字PCR进行的表达模式分析表明，（）在整个发育阶段和不同成虫组织中的表达水平各不相同，在四龄幼虫和血淋巴中表达水平最高。CmdsRNase可以降解dsRNA以降低（）中RNAi的效率。（来自）与CmdsRNase的共沉默显著影响了（）的生长发育，从而提高了RNAi效率，提高了27.17%。这些发现将有助于通过RNAi对（）和其他鳞翅目害虫进行绿色防控。

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