胡萝卜肉桂醇脱氢酶基因的克隆及其对非生物胁迫的响应

Cloning and Expression Profile Analysis of the Gene Encoding Cinnamyl Alcohol Dehydrogenase under Abiotic Stress in Carrot

该实验采用RT-PCR技术,从胡萝卜‘黑田五寸’中克隆获得了编码肉桂醇脱氢酶(Cinnamyl alcohol dehydrogenase,CAD)的基因DcCAD。DcCAD序列长1 074bp,编码357个氨基酸。亲水/疏水性分析表明,DcCAD属于亲水性蛋白。系统进化分析显示,DcCAD与番茄CAD(XP_010314515.1)亲缘关系最近。荧光定量PCR结果显示,DcCAD基因在胡萝卜根、叶片和叶柄中的表达量差异显著,其相对表达量为叶片>根>叶柄。DcCAD基因对高温(38℃)、低温(4℃)、干旱(20%PEG)和盐(0.2mol·L-1 NaCl)胁迫均有响应,尤其对高温胁迫和低温胁迫响应明显,而且高温处理后1h和低温处理后2h表达量最高。研究推测,DcCAD基因对胡萝卜抗逆性具有一定的作用。

Here ,the gene encoding DcCAD were cloned from carrot cultivar ‘Kurodagosun’ with RT-PCR method .The length of DcCA D was 1 074 bp ,and encoding 357 amino acids .The DcCAD may belong to hydrophilic protein . Phylogenetic analysis showed that DcCAD closed to CAD from tomato （XP_010314515 .1） .Quantitative real-time PCR analysis showed that the expression profiles of the DcCA D gene are significantly different in different tissues （root ,leaf blade ,and petiole） of carrot .Relative expression level of the DcCA D gene is leaf blade〉 root〉 petiole .The DcCA D gene is responsive to various abiotic stresses ,such as heat （38 ℃） ,cold （4 ℃） ,drought （20% PEG） and salt （0 .2 mol？L -1 NaCl） .The ex-pression level was significantly increased after high temperature or low temperature treatments ,respective-ly .The expression levels peaked at 1 h heat treatment and 2 h cold treatment ,respectively .These results suggested that DcCA D gene participate in the response process of carrot under the abiotic stress .