中华稻蝗几丁质合成酶1基因的mRNA表达及功能

doi:10.3864/j.issn.0578-1752.2012.05.007

摘要/Abstract

摘要： 【目的】研究中华稻蝗（Oxya chinensis（Thunberg））几丁质合成酶1（CHS1）基因的表达特性及其在生长发育过程中的作用，从而为实现基于RNAi的蝗虫有效控制提供理论依据。【方法】根据已知中华稻蝗几丁质合成酶1基因（OcCHS1）保守区域部分cDNA序列（GenBank登录号：HM214491）设计特异性表达引物，运用RT-PCR和qPCR研究时空表达特性；采用RNA干扰技术研究其生物学功能。【结果】OcCHS1在中华稻蝗各龄期都有表达，其体表为高表达，其次是气管，其它组织部位表达量低。RNA干扰试验发现，4龄第4天若虫注射OcCHS1的双链RNA（dsRNA）后，与对照组相比，OcCHS1 mRNA表达量被沉默了70.8%，稻蝗出现蜕皮时间延迟、不能完成蜕皮或腹部皱缩死亡等现象，注射dsOcCHS1组死亡率为85.2%，与对照组（7.4%）相比差异显著。【结论】几丁质合成酶1对中华稻蝗的生长发育具有重要作用，其主要参与体表和气管几丁质的合成。采用RNA干扰技术使该基因沉默后可导致中华稻蝗死亡。

关键词: 中华稻蝗, 几丁质合成酶基因1, mRNA表达, RNA干扰

Abstract: 【Objective】The objective of this study is to analyze the mRNA expression characteristics and physiological function of chitin synthase1 gene (OcCHS1) from the Chinese rice grasshopper, Oxya chinensis (Thunberg), and to provide a theoretical basis for effective pests control based on RNAi.【Method】Based on the known partial conservative cDNA fragment of OcCHS1 (GenBank accession number: HM214491), primers were designed for mRNA expression analysis. The developmental and tissue-specific expression patterns of OcCHS1 were determined by RT-PCR and qPCR. RNAi experiment was performed to explore the physiological function. 【Result】 OcCHS1 was expressed at all the developmental stages. Tissue-specific analysis showed that OcCHS1 was highly expressed in the integument, followed in the trachea, and lowest in other tissues. RNAi-based gene silencing experiment showed that the fourth day of the 4th instar nymphs injected by OcCHS1-speci?c dsRNA could repress the transcript levels of OcCHS1 by 70.8% compared to those in the control nymphs. Moreover, dsRNA injections signi?cantly increased abnormalities, such as stunted development, uncompleted molting, crimpy abdomen and eventually died. Consequentially, 85.2% of mortalities were observed, while the mortality in the control nymphs was only 7.4%. 【Conclusion】OcCHS1 is essential for development and mainly responsible for chitin synthesis in cuticle and trachea of O. chinensis. Moreover, silencing of chitin synthase 1 gene by RNAi is lethal to O. chinensis.

Key words: Oxya chinensis (Thunberg), chitin synthase 1 gene, mRNA expression, RNA interference

余志涛, 刘晓健, 马恩波, 郭亚平, 张建珍. 中华稻蝗几丁质合成酶1基因的mRNA表达及功能[J]. 中国农业科学, 2012, 45(5): 877-884.

YU Zhi-Tao, LIU Xiao-Jian, MA 恩Bo, GUO Ya-Ping, ZHANG Jian-Zhen. mRNA Expression and Physiological Function of Chitin Synthase1 from Oxya chinensis (Thunberg)[J]. Scientia Agricultura Sinica, 2012, 45(5): 877-884.

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