滞育七星瓢虫抑制性消减杂交文库的构建及分析

doi:10.16409/j.cnki.2095-039x.2016.01.005

中国生物防治学报 ›› 2016, Vol. 32 ›› Issue (1): 33-39.DOI: 10.16409/j.cnki.2095-039x.2016.01.005

滞育七星瓢虫抑制性消减杂交文库的构建及分析

蒋莎^1,2, 黄凤霞², 齐晓阳¹, 任小云², 陈红印², 黄建¹, 张礼生²

1. 福建农林大学植物保护学院, 福州 350002;
2. 中国农业科学院植物保护研究所/农业部作物有害生物综合治理重点实验室/中美生物防治实验室, 北京 100193

收稿日期:2015-03-18 出版日期:2016-02-08 发布日期:2016-02-22
通讯作者: 张礼生, 黄建
作者简介:蒋莎(1989-),女,硕士研究生,E-mail:jiangshajs@126.com
基金资助:
国家公益性行业(农业)科研专项(201103002);农业部"948"项目(2011-G4)

Construction and Analysis of Suppression Subtractive Hybridization cDNA Library of the Diapause Coccinella septempunctata L.

JIANG Sha^1,2, Huang Fengxia², Qi Xiaoyang¹, Ren Xiaoyun², CHEN Hongyin², HUANG Jian¹, ZHANG Lisheng²

1. College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou 350002, China;
2. Key Laboratory of Integrated Pest Management in Crops, Ministry of Agriculture/Institute of Plant Protection, Chinese Academy of Agricultural Sciences/Sino-American Biological Control Laboratory, Beijing 100193, China

Received:2015-03-18 Online:2016-02-08 Published:2016-02-22

摘要/Abstract

摘要： 七星瓢虫是优良的天敌昆虫,可凭滞育特性抵御逆境胁迫,提高种群生存能力。为研究七星瓢虫的滞育分子机制,构建抑制性消减杂交文库,经反向Northern斑点杂交技术进一步筛选,得到231个差异点。blastx/n比对后,正、反向文库分别得到差异表达基因64和54个,GO分类和PATHWAY分析表明,差异表达基因参与的生物过程涉及滞育的信号传导、生物合成、初级代谢和次级代谢等。文库中发现多种基因,为进一步筛选、克隆、分析滞育相关基因提供线索。着重讨论了与抗寒性相关的28 kD抗干燥应激蛋白、卵巢发育相关的Cullin 3蛋白,能量代谢相关的异柠檬酸脱氢酶的生物学功能,进而推测其与七星瓢虫滞育的相关性。

关键词: 七星瓢虫, 抑制性消减杂交, 差异表达基因, 反向Northern 斑点杂交, 滞育

Abstract: The ladybird Coccinella septempunctata L., an excellent insect natural enemy, enter diapause to avoid unfavorable environmental conditions, which offers a mechanism for a survival. In this study, suppression subtractive hybridization (SSH) library and reverse Northern hybridization blot were employed to screen differentially expressed genes in the diapause insects and 231 differentially dots were obtained. Using an approach of blastx/n alignment, 64 and 54 differentially expressed genes were identified in forward and reverse library, respectively. These genes were mainly involved in signal transduction, biosynthesis, primary metabolism and secondary metabolism. Multiple genes found in the library provide references for further screening, cloning, and analysis of diapause-associated genes. The biological functions of 28 kD desiccation stress protein associated with cold hardiness, Cullin 3 related to the development of ovary and isocitrate dehydrogenase involved in energy metabolism were discussed in particular and their relatedness with diapause in C. septempunctata was reviewed.

Key words: Coccinella septempunctata, suppression subtractive hybridization, differentially expressed genes, reverse Northern dot-blotting, diapause

中图分类号:

S476.2

蒋莎, 黄凤霞, 齐晓阳, 任小云, 陈红印, 黄建, 张礼生. 滞育七星瓢虫抑制性消减杂交文库的构建及分析[J]. 中国生物防治学报, 2016, 32(1): 33-39.

JIANG Sha, Huang Fengxia, Qi Xiaoyang, Ren Xiaoyun, CHEN Hongyin, HUANG Jian, ZHANG Lisheng. Construction and Analysis of Suppression Subtractive Hybridization cDNA Library of the Diapause Coccinella septempunctata L.[J]. journal1, 2016, 32(1): 33-39.

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滞育七星瓢虫抑制性消减杂交文库的构建及分析

Construction and Analysis of Suppression Subtractive Hybridization cDNA Library of the Diapause Coccinella septempunctata L.

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