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棉花学报 ›› 2016, Vol. 28 ›› Issue (6): 547-554.doi: 10.11963/issn.1002-7807.201606004

• 研究与进展 • 上一篇    下一篇

棉花抗黄萎病相关基因的序列和表达分析

赵君1(),徐剑文1,刘剑光1,吴巧娟1,肖松华1,**(),赵亮1,袁有禄2,**()   

  1. 1.江苏省农业科学院经济作物研究所/农业部长江下游棉花和油菜重点实验室,江苏 南京 210014
    2.中国农业科学院棉花研究所/棉花生物学国家重点实验室,河南 安阳 455000
  • 收稿日期:2016-01-04 出版日期:2016-11-15 发布日期:2016-11-15
  • 通讯作者: 肖松华,袁有禄 E-mail:sxzhaojun88@aliyun.com;njxsh@sina.com;yuanyl@cricaas.com.cn
  • 作者简介:赵君(1982―),男,博士,副研究员,sxzhaojun88@aliyun.com
  • 基金资助:
    国家自然科学基金(31401727);江苏省重点研发项目(BE2015353);江苏省自然科学基金(BK20130699);棉花生物学国家重点实验室开放课题(BC2015A07);国家自然科学基金青年项目配套资金(0120656100ZX)

Cloning and Expression Analysis of Resistance-related Genes to Verticillium Wilt in Cotton

Zhao Jun1(), Xu Jianwen1, Liu Jianguang1Wu Qiaojuan1, Xiao Songhua1,*(), Zhao Liang1YuanYoulu2,*()   

  1. 1. Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences/Key Laboratory of Cotton and Rapeseed, Ministry of Agriculture, Nanjing, Jiangsu 210014, China
    2. Institute of Cotton Research, Chinese Academy of Agricultural Sciences/State Key Laboratory of Cotton Biology, Anyang, Henan 455000, China
  • Received:2016-01-04 Online:2016-11-15 Published:2016-11-15
  • Contact: Songhua Xiao,Youlu Yuan E-mail:sxzhaojun88@aliyun.com;njxsh@sina.com;yuanyl@cricaas.com.cn

摘要: 黄萎病是棉花生产的主要病害,克隆抗病基因是培育棉花抗黄萎病品种的关键。本文根据前期的定位结果,结合棉花基因组测序信息,提取定位区段内基因组序列,预测获得63个基因。Gene Ontology分析表明63个基因参加多种生物进程,其中6个基因参与植物抗逆进程。根据Gene Ontology分析和前人研究结果,选择15个基因进行序列分析。启动子序列分析表明启动子区域包含各种抗逆的调控元件,其中6个基因包含了W-box元件。对海7124和苏棉8号棉花幼苗进行黄萎病菌处理,取病菌处理后不同时期的根,选择14个基因进行表达分析,结果表明在黄萎病菌处理后,有8个基因表达出现变化,其中在海7124和苏棉8号之间表达差异最大的基因是跨膜蛋白(Transmembrane protein 214-a isoform 1)、 细胞色素P450(Cytochrome p450)和Udp-糖基转移酶UGT89A2(Udp-glycosyl transferase 89a2-like)基因。本研究为抗病基因克隆提供了候选基因。

关键词: 棉花; 黄萎病; 抗病基因; 克隆

Abstract:

Verticillium wilt is a major limiting factor in cotton production, and the key of breeding resistant varieties by genetic engineering is to clone resistance genes. In this study, based on the cotton genomic sequence and previous Verticillium wilt-resistance gene predictions, we extracted the target sequence and predicted that it contained 63 genes. The gene Ontology showed that 63 genes participated in a variety of biological processes, and 6 of them were involved in plant stress responses. According to the results of the Gene Ontology analysis and previous studies, we selected 15 genes as research subjects. A promoter sequence analysis showed that the promoter regions of 15 genes contained a variety of cis-acting elements, and 6 genes contained W-box components. Using the root of Gossypium barbadense L. cv. Hai7124 and Gossypium hirsutum L. cv. Sumian 8 seedlings treated with Verticillium dahliae for different times, we analyzed the expression profiles of 14 genes. The expression levels of eight genes were induced and changed after treatment with V. dahliae, and the expression differences of transmembrane protein 214-a isoform 1, Cytochrome P450 and Udp-glycosyl transferase 89a2-like were the most significant between G. barbadense L. cv. Hai7124 and G. hirsutum L. cv. Sumian 8. The study provides candidate Verticillium wilt-resistance genes for cloning in the future.

Key words: cotton; Verticillium dahliae; resistance gene; cloning