烟草BRANCHED1-Like基因的克隆及表达分析  

Molecular Cloning and Expression Analysis of BRANCHED1-Like Gene in Common Tobacco

作　　者：陈雅琼[1,2] 孙亭亭[1,2] 陈蕾[1,2] 丁安明[1] 陈涣[1,2] 孙玉合[1] 

机构地区：[1]中国农业科学院烟草研究所/烟草行业烟草基因资源利用重点实验室,青岛266100 [2]中国农业科学院研究生院,北京100081

出　　处：《植物遗传资源学报》2015年第6期1321-1329,共9页Journal of Plant Genetic Resources

基　　金：国家高科技研究发展计划("863"计划)(2012AA021801)

摘　　要：TCP转录因子家族CYC/TB1簇中成员BRC1在调控植物侧枝发育的过程中发挥重要作用。本研究利用电子克隆结合RT-PCR方法从普通烟草(Nicotiana tabacum)中克隆获得4个Nt BRC1-Like基因,分别命名为Nt BRC1-Like1、Nt BRC1-Like2、Nt BRC1-Like3和Nt BRC1-Like4。4条基因的CDS序列长度分别为1140 bp、1062 bp、987 bp和1041 bp,分别编码由379、353、328和346个氨基酸组成的蛋白,这4个蛋白与CYC/TB1簇中其他成员具有较高相似性。蛋白质保守结构域分析表明,4个Nt BRC1-Like基因编码的蛋白除具有TCP核心结构域外,还具有CYC/TB1簇蛋白特有的R结构域。系统进化树分析表明,Nt BRC1-Like蛋白都是TCP家族CYC/TB1簇中的成员,且4个Nt BRC1-Like蛋白可以被分成2组,Nt BRC1-Like1和Nt BRC1-Like4为一组,Nt BRC1-Like2和Nt BRC1-Like3为另一组。荧光定量PCR分析发现,4个烟草Nt BRC1-Like基因的表达呈现明显的组织特异性,且其表达模式可分为2种:Nt BRC1-Like1和Nt BRC1-Like4的表达模式相似,在腋芽中有极高的表达;Nt BRC1-Like2和Nt BRC1-Like3的表达模式十分相似,在叶和腋芽中的表达量都显著高于其他组织。推测烟草BRC1基因可能存在类似番茄BRC1基因在功能上保守与进化的情况。BRC1, a member of CYC/TB1 subclass in TCP transcription factor family, plays an important role in the regulation of shoot branching. Four new NtBRC1-Like genes, designated NtBRC1-Likel, NtBRC1-Like2, NtBRC1- Like3 and NtBRC1-Like4, were isolated from Nicotiana tabacum using silico cloning strategy combined RT-PCR methods. The length of CDS sequences of four NtBRC1-Like genes were 1140 bp, 1062 bp ,987 bp and 1041 bp ,respectively. The deduced protein sequences contained 379,353,328 and 346 amino acids, respectively, and exhibited high homology with other members of CYC/TB1 subclass. The analysis of conserved domain showed that besides TCP core domain,four NtBRC1-Like proteins also had the R domain specific to CYC/TB1 proteins. The phylogenetic tree showed that four NtBRC1-Like proteins belonged to CYC/TB1 subclass of TCP family,furthermore,four NtBRC1-Like proteins could be divided into two groups,one was NtBRC1-Likel/4,another was NtBRC1-Like2/3. Real-time quantitative PCR assay indicated that the expression profile of NtBRC1-Like genes exhibited significant tis- sue specificity. The expression level of NtBRC1-Likel/4 in axillary bud was the highest, however,NtBRC1-Like2/3 expressed higher in leaf and axillary bud. These results demonstrated that the functions of NtBRCls in tobacco might be differentiated just like in tomato.

关 键 词：烟草 BRC1基因 侧枝形成 基因克隆 基因表达分析 

分 类 号：S572[农业科学—烟草工业] Q943.2[农业科学—作物学]