巴拉圭瓜多竹CAD基因的克隆与分析  

Cloning and Sequence Analysis of Cinnamoyl Alcohol Dehydrogenase Gene from Guadua paraguayana

作　　者：朱金鑫[1,2] 孙金金[1,2] 原晓龙[1,3] 王娟[1,3] 杨宇明[1,3] 王毅[1,3] 

机构地区：[1]国家林业局重点开放性实验室云南珍稀濒特森林植物保护和繁育实验室,云南省森林植物培育与开发利用重点实验室,云南昆明650201 [2]西南林业大学,云南昆明650224 [3]云南省林业科学院,云南昆明650201

出　　处：《西部林业科学》2017年第1期6-11,16,共7页Journal of West China Forestry Science

基　　金：云南省对外科技合作计划(2014IA3);云南省基础研究重点项目(2013FA054);云南省中青年学术技术带头人后备人才培养项目(2010CI016);国家林业局云南珍稀濒特森林植物繁育和保护重点实验室开发基金共同资助

摘　　要：肉桂醇脱氢酶(cinnamyl alcohol dehydrogenase,CAD,EC1.1.1.195)是木质素生物合成途径中的最后关键酶,在木质素的生物合成中发挥关键作用。本研究依据转录组数据设计特异性引物,采用RT-PCR方法成功地从巴拉圭瓜多竹中克隆得到一个全新CAD基因的c DNA全长序列,命名为GPCAD(Gene Bank登录为KJ784465)。序列分析结果表明,GPCAD基因片段包含完整的c DNA开放阅读框(ORF),序列全长1 080bp,编码含有359个氨基酸残基的蛋白质。Blast比对结果显示该蛋白质属于CAD家族蛋白;系统进化分析结果显示巴拉圭瓜多竹与禾本科植物亲缘关系较近;荧光定量PCR技术检测结果显示GPCAD在茎秆中表达量最高,叶中的表达量最低。本研究对巴拉圭瓜多竹CAD基因进行克隆与分析,为深入研究巴拉圭瓜多竹木质素代谢途径相关基因以及通过分子生物学手段对木质素的含量和单体比例进行调控提供科学依据。Cinnamyl alcohol dehydrogenase （CAD, EC1.1.1. 195） is the last key enzyme in the specific pathway of lignin biosynthesis, and it plays an key role in the biosynthesis of lignin. In this study, a full-length eDNA sequences of a new CAD genes was successfully cloned from G. paraguayana by RT-PCR with the specific primers designed based on transcriptome datas, named GPCAD （GeneBank, KJ784465 ）. Sequences analysis showed that cDNA fragment of GPCAD gene containing the complete open reading frame （ ORF）, the full-length sequence of 1080 bp, encoded a protein with 359 amino acid. Blast comparison showed that the protein belonged to the CAD family; phylogenetic tree showed Guadua paraguayana had close genetic relationship with the plants belongings to Graminales; RT-PCR showed GPCAD had the highest expression in culm, the lowest in leaf. In this study, the CAD gene cloned from G. paraguayana provide a scientific basis for further study about related genes in Lignin metabolic pathways and regulation of lignin content and proportion by molecular methods.

关 键 词：巴拉圭瓜多竹 木质素 肉桂醇脱氢酶 基因克隆与分析 

分 类 号：S795.9[农业科学—林木遗传育种]