Cloning of ERF Transcription Factor Gene and Its Response Analysis Induced by Abiotic Stress in Sugar Beet
孙亚卿 1米翔宇 1王彤 1王妍 1李宁宁 1张少英 1李国龙1
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作者信息
1. 内蒙古农业大学农学院,呼和浩特,010019
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摘要
ERF(Ethylene-responsive element binding factors)是调控植物生长发育和响应胁迫的重要转录因子,在植物应对生物及非生物胁迫反应、调控胁迫相关功能基因的表达、提高植物的抗逆性中起着重要的作用.分析甜菜ERF转录因子基因在非生物胁迫下的表达规律,旨在为深入研究ERF转录因子在甜菜逆境胁迫应答中的作用机制提供理论依据.本研究以高糖型甜菜品系'BS02'为试材,利用RT-PCR方法克隆得到Bv_ammr基因,并对其进行生物信息学分析.该基因CDS区长906 bp,编码301个氨基酸.蛋白质分子量为33.19 kD,理论等电点为8.61,其二级结构以无规则卷曲和α-螺旋为主,具有一个AP2保守结构域;氨基酸序列的同源性和进化树分析显示其编码蛋白与马兜铃菌毛所编码的蛋白亲缘关系较近.采用实时荧光定量PCR方法观测非生物胁迫下该基因表达模式,结果表明,Bv_ammr基因对低温、高温、干旱、盐、ABA等非生物胁迫有不同程度响应.
Abstract
ERF(Ethylene-resp active element binding factors)is an important transcription factor that regulates plant growth and response to stress,and plays an important role in plant response to biological and abiotic stress,regulating the expression of stress-related functional genes,and improving plant resistance.The expression of ERF transcription factor gene in sugar beet under abiotic stress was analyzed to provide theoretical basis for studying its mechanism in stress regulation.In this study,Bv_ammr was cloned by RT-PCR from sugar beet,which CDS region length was 906 bp long,encoding 301 amino acids.Bioinformatics analysis showed that the molecular weight of the protein was 33.19 kD,the theoretical isoelectric point was 8.61,and its secondary structure was irregular curl and α-helix-dominated,with an AP2 conserved domain.The homology and phylogenetic tree analysis of amino acid sequence showed that its encoded protein was close to the protein encoded by A ristolochia pili.Real-time fluorescence quantitative PCR was used to observe the expression pattern of the gene under abiotic stress.The results showed that Bv_ammr gene had different responses to abiotic stresses such as low temperature,high tem-perature,drought,salt,strong light and weak light.
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