棉花乙醇酸氧化酶(Glycolate oxidase)基因的克隆及表达分析  

Cloning and Expression Analysis of Glycolate oxidase in Cotton(Gossypium hirsutum)

作　　者：张雪 董丽君[1] 封润霞 姚莹 刘建凤[1] 张书玲[1] Zhang Xue;Dong Lijun;Feng Runxia;Yao Ying;Liu Jianfeng;Zhang Shuling(College of Life Scicnce,Hebei Universiy,Baoding,071002)

机构地区：[1]河北大学生命科学学院,保定071002

出　　处：《分子植物育种》2020年第22期7309-7316,共8页Molecular Plant Breeding

基　　金：河北大学研究生创新资助项目(hbu2019ss027)资助。

摘　　要：黄萎病是造成棉花产量和品质严重下降的重要病害,生产上至今无法根治。棉花抗黄萎病分子机制不明是阻碍棉花分子育种的主要原因。乙醇酸氧化酶(glycolate oxidase, GOX)是植物中产生ROS主要来源之一,在植物抗病性中起着至关重要的作用。本研究从棉花基因组中鉴定了11个GOX家族成员,并克隆了其中一个成员GHD01G0345,其开放阅读框(open reading frame, ORF)为1 110 bp,编码369个氨基酸。该基因编码氨基酸序列中包含一个完整的α-羟基酸氧化-FMN域和关键的催化活性氨基酸。结果表明,当陆地棉受到黄萎病菌侵染12 h时,出现活性氧积累和过氧化氢(H2O2)含量升高,同时GHD01G0345基因表达量也达到最大值。启动子顺式作用元件分析发现GHD01G0345启动子中除了大量的光反应元件外,还有参与水杨酸(salicylic acid, SA)反应的顺式作用元件。当SA诱导后,GHD01G0345表达模式与黄萎病菌胁迫后相似,其表达量在6 h达到最大值。由此推测,棉花体内SA水平升高会使GHD01G0345基因表达上调,导致ROS的大量产生,进而介导多种应答反应,最终使棉花获得对黄萎病菌的抗性。Verticillium wilt is an important disease that causes serious decline of cotton yield and quality, which can not be cured in production up to now. The unclear molecular mechanism of cotton resistance to Verticillium wilt is the main reason that hinders cotton molecular breeding. Glycolate oxidase(GOX) is one of the main sources of ROS in plants, which plays an important role in plant disease resistance. In this study, 11 members of GOX family were identified from cotton genome, and one of them, GH_D01 G0345, was cloned. The open reading frame of this gene was 1 110 bp, encoding 369 amino acids. The amino acid sequence of the gene contained an alpha_hydroxyacid_oxid_FMN domain and key catalytic active amino acids. The results indicated when cotton was infected by Verticillium dahliae for 12 h, the accumulation of active oxygen and the content of hydrogen peroxide(H2 O2) increased, and the expression of GH_D01 G0345 gene also reached the maximum. In addition to a large number of photoreactive elements, there are also cis acting elements involved in the reaction of salicylic acid(SA) in the promoter of GH_D01 G0345. After SA induction, the expression pattern of GH_D01 G0345 was similar to that of Verticillium dahliae, and its expression reached the maximum value at 6 h. It can be concluded that the increase of SA level in cotton will increase the gene expression of GH_D01 G0345, lead to the production of ROS,and then mediate a variety of response reactions, and finally make cotton obtain resistance to Verticillium wilt.

关 键 词：陆地棉 乙醇酸氧化酶 克隆 表达分析 黄萎病 

分 类 号：S562[农业科学—作物学]