丹参咖啡酸-O-甲基转移酶基因(SmCOMT1)的克隆及其分析  

Cloning and Analysis of Caffeic Acid O-methyltransferase Gene(SmCOMT1) from Salvia miltiorrhiza Bge.

作　　者：宋银[1] 王东浩[1] 吴锦斌[1] 周露[1] 王国栋[1] 王喆之[1] 

机构地区：[1]药用资源与天然药物化学教育部重点实验室,西北濒危药材资源开发国家工程实验室,陕西师范大学生命科学学院,西安710062

出　　处：《植物研究》2012年第4期437-443,共7页Bulletin of Botanical Research

基　　金：国家自然科学基金项目(31000134)资助

摘　　要：依据丹参转录组数据库得到的咖啡酸-O-甲基转移酶基因序列设计特异性引物,采用RT-PCR方法从丹参分离得到一个新的COMT基因,命名为SmCOMT1(GenBank注册号为JF693491)。该基因cDNA全长1 158 bp,包含一个长为1 095 bp的开放阅读框,编码364个氨基酸。SmCOMT1 gDNA序列长2 275 bp,包含4个外显子和3个内含子。序列分析结果表明,SmCOMT1编码的多肽具有COMT的序列保守元件,与同科植物罗勒COMT编码的多肽高度同源,同源性达到89%。系统进化树分析表明,SmCOMT1与双子叶植物的COMT亲缘关系较近。qRT-PCR结果表明,SmCOMT1基因在丹参不同组织器官中差异表达,其中茎中的表达量最高,并且其表达受茉莉酸甲酯和病原菌的诱导,显示SmCOMT1基因可能在植物防御反应中发挥作用。According to the sequencing result of caffeic acid O-methyltransferase from Salvia miltiorrhiza transcriptome database analysis, its specific primers were designed. By RT-PCR method, a novel COMT gene was i-solated from S. miltiorrhiza, and named as SmCOMT1 （ Genebank accession number： JF693491 ）. SmCOMT1, with full-length cDNA of 1 158 bp, includes an open reading frame of 1 095 bp which encodes a 364 amino acids polypeptide. Furthermore, a length of 2 275 bp sequence was also cloned by PCR from genomic DNA of S. miltiorrhiza. The genomic DNA of SmCOMT1, aligned with cDNA, contains four exons and three introns in the encoding region. The results of amino acid sequence analysis shows that deduced amino acid polypeptide contains all the conserved element of COMT family and it is highly homologous to COMT proteins from the same family of Ocimum basilicum with 89% identity. Phylogenetic tree analysis also indicates that SmCOMT1 is more related to the genetic relationship of COMT in dicotyledonous plants. Quantitative RT-PCR analysis revealed that SmCOMT1 was expressed in different organs, and was highly expressed in stem, and could be induced by methyl jasmonate （MeJA） and pathogen. These results showed that SmCOMT1 might be pathogen-responsive gene in plant defenses.

关 键 词：丹参 咖啡酸-O-甲基转移酶 基因克隆 qRT—PCR 

分 类 号：S567.2[农业科学—中草药栽培]