茶树黄酮合成酶Ⅱ基因全长cDNA序列的克隆和实时荧光定量PCR检测  

Molecular Cloning and Real-Time PCR Analysis of Flavone SynthaseⅡ Gene Full-length cDNA from the Tea Plant

作　　者：乔小燕[1,2] 马春雷[1] 陈亮[1] 

机构地区：[1]中国农业科学院茶叶研究所茶树资源与改良研究中心,国家茶树改良中心,浙江杭州310008 [2]广东省农业科学院茶叶研究所,广东广州510640

出　　处：《茶叶科学》2009年第5期347-354,共8页Journal of Tea Science

基　　金：国家863计划(2006AA10Z171);“现代农业产业技术体系建设专项资金资助”内容之一

摘　　要：利用RT-PCR和RACE技术,克隆得到茶叶中合成黄酮类化合物的关键酶基因—黄酮合成酶Ⅱ,该基因属于细胞色素P450家族,在GenBank登录号为FJ169499。黄酮合成酶ⅡcDNA全长序列1824bp,其中1605bp为编码区,编码534个氨基酸,分子量为60.4kD。通过SYBRGreenI实时荧光定量PCR检测,发现黄酮合成酶Ⅱ在茶树一芽二叶春梢、当季成熟叶、花瓣、花蕊、种子中都有表达,成熟叶的表达量显著高于其他四种组织。Using RT-PCR and RACE techniques, flavone synthaseⅡ （FS Ⅱ） gene, the key enzyme in synthesize flavone, was cloned in the tea plant. The FS Ⅱ belongs to cytoehrome P450 superfamily. The GenBank Accession No. is FJ169499 in the NCBI. The FS Ⅱ gene eDNA had 1 824 bp in full-length with 1 605 bp in coding region, encoding 534 amino acids. The putative molecular weight was 60.4 kD. The expression levels of flavone synthase Ⅱ gene in spring ＇two and a bud＇, mature leaves, petals, stamen and pistils, and seeds of the tea plant were assessed using SYBR Green Ⅰ Real-time PCR. The expression in mature leaves was significantly higher than that in the other four tissues.

关 键 词：茶树 黄酮合成酶Ⅱ 实时荧光PCR 细胞色素P450家族 

分 类 号：S571.1[农业科学—茶叶生产加工] Q523[农业科学—作物学]