一个亚洲棉MYB家族新基因的克隆及特征分析

doi:10.3864/j.issn.0578-1752.2013.01.023

中国农业科学 ›› 2013, Vol. 46 ›› Issue (1): 195-204.doi: 10.3864/j.issn.0578-1752.2013.01.023

一个亚洲棉MYB家族新基因的克隆及特征分析

杨召恩, 杨作仁, 刘坤, 刘传亮, 张朝军, 李付广

1.中国农业科学院棉花研究所/棉花生物学国家重点实验室/农业部棉花遗传改良实验室，河南安阳 455000
2.周口师范学院植物遗传与分子育种重点实验室，河南周口 466000

收稿日期:2012-09-18 出版日期:2013-01-01 发布日期:2012-11-13
通讯作者: 通信作者李付广，Tel：0372-2562204；E-mail：aylifug@163.com
作者简介:杨召恩，E-mail：yangzhaoen0925@126.com
基金资助:
国家杰出青年基金项目“棉花高频再生性状的遗传与应用研究”（31125020）

Cloning and Characterization of a Noval Gene of MYB Family from Gossypium arboreum L.

YANG Zhao-恩, YANG Zuo-Ren, LIU Kun, LIU Chuan-Liang, ZHANG Chao-Jun, LI Fu-Guang

1.Cotton Research Institute, Chinese Academy of Agricultural Sciences/State Key Laboratory of Cotton Biology/Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Anyang 455000, Henan
2.Key Laboratory of Plant Genetics and Molecular Breeding of Zhoukou Normal University, Zhoukou 466000, Henan

Received:2012-09-18 Online:2013-01-01 Published:2012-11-13

摘要/Abstract

摘要： 【目的】克隆亚洲棉MYB家族的一个新基因（GaMYB2），研究其表达模式，分析其预期编码蛋白。【方法】利用RT-PCR和RACE技术克隆亚洲棉MYB2的cDNA序列全长；应用生物信息学软件分析该基因及预期编码蛋白的特征；采用实时荧光定量PCR技术对该基因的组织特异性和PEG6000模拟干旱胁迫处理下的表达模式进行分析。【结果】从亚洲棉(Gossypium arboreum L.)中克隆了MYB家族的一个新基因MYB2，该基因全长1 117 bp,ORF全长840 bp，编码279个氨基酸，含有MYB保守域，氨基酸的BALSTP分析表明GaMYB2氨基酸序列与已报道的水稻（BAA23338.1）、小麦(AAT37168.1)等的序列有40.50%到68.20%的相似性。亚细胞定位表明该基因在细胞核中表达，实时荧光定量PCR结果表明该基因在根和花中的表达量较高，并且响应17%的PEG6000模拟干旱胁迫处理，上调表达。【结论】GaMYB2是亚洲棉MYB家族的一个新基因，并认为该基因可能在棉花调控干旱胁迫的生理适应过程中发挥重要作用。

关键词: 亚洲棉 , GaMYB2 , 干旱模拟 , 上调表达

Abstract: 【Objective】The objective of this study is to clone a novel gene of MYB family (GaMYB2) from Gossypium arboreum L., to analyze its expression patterns, and to characterize the deduced protein of this gene. 【Method】The full length of GaMYB2 was cloned by RACE and RT-PCR techniques, the gene sequence obtained and the putative amino acid sequence was analyzed by bioinformatics software. Its expression patterns were analyzed by real-time PCR under normal and PEG6000 stress respectively，which simulated drought tolerance.【Result】The GaMYB2 full length cDNA sequence is 1 117 bp, and the ORF is 840 bp, which encode a deduced protein including 279 amino acid residues. Blastp reseach revealed 40.50% to 68.20% similarities between GaMYB2 and other reported species, such as Oryza sativa (BAA23338.1) and Triticum aestivum (AAT37168.1). Subcellular localization showed that it expressed possibly in cell nucleus. The experiment results of real time PCR exhibited that GaMYB2 could be induced by 17% PEG6000, and had a higher expression level in root and flower than in other tissues. 【Conclusion】The GaMYB2 , cloned for the first time from G. arboreum L., is a new transcription factor of MBY family, and may play an important role in plant tolerance to drought stress.

Key words: Gossypium arboreum L. , GaMYB2 , drought tolerance simulation , up-regulate expression

杨召恩, 杨作仁, 刘坤, 刘传亮, 张朝军, 李付广. 一个亚洲棉MYB家族新基因的克隆及特征分析[J]. 中国农业科学, 2013, 46(1): 195-204.

YANG Zhao-恩, YANG Zuo-Ren, LIU Kun, LIU Chuan-Liang, ZHANG Chao-Jun, LI Fu-Guang. Cloning and Characterization of a Noval Gene of MYB Family from Gossypium arboreum L.[J]. Scientia Agricultura Sinica, 2013, 46(1): 195-204.

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一个亚洲棉MYB家族新基因的克隆及特征分析

Cloning and Characterization of a Noval Gene of MYB Family from Gossypium arboreum L.

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