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利用 RNA 干扰技术对调控水稻褐飞虱几丁质代谢途径的三个海藻糖酶基因进行功能表征。

Functional characterization of three trehalase genes regulating the chitin metabolism pathway in rice brown planthopper using RNA interference.

机构信息

Hangzhou Key Laboratory of Animal Adaptation and Evolution, College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou, Zhejiang, 310036, China.

出版信息

Sci Rep. 2016 Jun 22;6:27841. doi: 10.1038/srep27841.

Abstract

RNA interference (RNAi) is an effective gene-silencing tool, and double stranded RNA (dsRNA) is considered a powerful strategy for gene function studies in insects. In the present study, we aimed to investigate the function of trehalase (TRE) genes (TRE 1-1, TRE 1-2, and TRE-2) isolated from the brown planthopper Nilaparvata lugens, a typical piercing-sucking insect in rice, and investigate their regulating roles in chitin synthesis by injecting larvae with dsRNA. The results showed that TRE1 and TRE2 had compensatory function, and the expression of each increased when the other was silenced. The total rate of insects with phenotypic deformities ranged from 19.83 to 24.36% after dsTRE injection, whereas the mortality rate ranged from 14.16 to 31.78%. The mRNA levels of genes involved in the chitin metabolism pathway in RNA-Seq and DGEP, namely hexokinase (HK), glucose-6-phosphate isomerase (G6PI) and chitinase (Cht), decreased significantly at 72 h after single dsTREs injection, whereas two transcripts of chitin synthase (CHS) genes decreased at 72 h after dsTRE1-1 and dsTREs injection. These results demonstrated that TRE silencing could affect the regulation of chitin biosynthesis and degradation, causing moulting deformities. Therefore, expression inhibitors of TREs might be effective tools for the control of planthoppers in rice.

摘要

RNA 干扰 (RNAi) 是一种有效的基因沉默工具,双链 RNA (dsRNA) 被认为是研究昆虫基因功能的有力策略。在本研究中,我们旨在研究从水稻上的典型刺吸式昆虫褐飞虱中分离出的海藻糖酶 (TRE) 基因 (TRE1-1、TRE1-2 和 TRE-2) 的功能,并通过注射 dsRNA 研究它们在几丁质合成中的调节作用。结果表明, TRE1 和 TRE2 具有代偿功能,当另一个基因沉默时,每个基因的表达都会增加。dsTRE 注射后,具有表型畸形的昆虫总比率从 19.83%到 24.36%不等,而死亡率从 14.16%到 31.78%不等。在 RNA-Seq 和 DGEP 中,参与几丁质代谢途径的基因,即己糖激酶 (HK)、葡萄糖-6-磷酸异构酶 (G6PI) 和几丁质酶 (Cht) 的 mRNA 水平在单次 dsTRE 注射后 72 小时显著降低,而 CHS 基因的两个转录本在 dsTRE1-1 和 dsTREs 注射后 72 小时降低。这些结果表明, TRE 沉默可能会影响几丁质生物合成和降解的调节,导致蜕皮畸形。因此, TREs 的表达抑制剂可能是控制水稻稻飞虱的有效工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/45b4/4916506/0f7b1b29c95c/srep27841-f1.jpg

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