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东亚飞蝗几丁质酶家族基因的表达特性与功能研究

李大琪 杜建中 张建琴 郝耀山 刘晓健 王亦学 马恩波 张建珍 孙毅

中国农业科学2011,Vol.44Issue(3):485-492,8.
中国农业科学2011,Vol.44Issue(3):485-492,8.DOI:10.3864/j.ssn.0578-1752.2011.03.007

东亚飞蝗几丁质酶家族基因的表达特性与功能研究

Study on Expression Characteristics and Functions of Chitinase Family Genes from Locusta migratoria manilensis (Meyen)

李大琪 1杜建中 2张建琴 1郝耀山 2刘晓健 1王亦学 2马恩波 1张建珍 1孙毅2

作者信息

  • 1. 山西大学应用生物学研究所,太原,030006
  • 2. 山西省农业科学院生物技术研究中心,太原,030031
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摘要

Abstract

[Objective] In order to provide important basis for effective pests control based on RNAi, the temporal and spatial expression characterizations and the biological functions of chitinase family genes from L. migratoria manilensis were investigated and chitinase genes causing locusts death during development were screened. [ Method ] Chitinase-like gene fragments were obtained from locustDB and the sequences of these genes were analyzed by bioinformatics methods. The first-strand cDNAs were synthesized using RNA isolated from the fourth day of different locust stages and the various tissues of the 5th nymphs and RT-PCR was carried out to analyze the gene expression patterns. Biological functions of LmCht6 that was highly expressed in the integument were studied by RNA interference. [ Result ] Seven chitinase-like gene fragments were gained from locustDB. The RT-PCR results showed that the temporal and spatial expression patterns among seven chitinase genes were significantly different and LmCht6 was highly expressed in the integument. RNA interference of LmCht6 showed that the corresponding transcript level was reduced in the nymphs after being injected with LmCht6 dsRNA compared with the control insects. The nymphs injected with LmCht6 dsRNA displayed slow development and failed to detach old cuticle during molting. Consequently, 72.2% of mortalities were observed. [ Conclusion ] More than one chitinase gene was present in locust and the temporal and spatial expression profiles of these genes were different.The RNAi results indicated that LmCht6 play an important role in the molting process of L. migratoria manilensis, and gene silencing can result in its ecdysis block and even death.

关键词

东亚飞蝗/几丁质酶基因/RT-PCR/RNA干扰

引用本文复制引用

李大琪,杜建中,张建琴,郝耀山,刘晓健,王亦学,马恩波,张建珍,孙毅..东亚飞蝗几丁质酶家族基因的表达特性与功能研究[J].中国农业科学,2011,44(3):485-492,8.

基金项目

国家自然科学基金(30970410)、山西省回国留学人员科研资助项目(2009)、国家转基因生物新品种培育重大科技专项(2008ZX08003-001)、国家公益性行业科技项目(200903021) (30970410)

中国农业科学

OA北大核心CSCDCSTPCD

0578-1752

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