花生C4H和ANR基因的克隆与表达研究  

CLONING AND EXPRESSION ANALYSIS OF C4H AND ANR GENES IN PEANUT(Arachis hypogaea L.)

作　　者：马敬[1,2,3] 苏磊[1,2,3] 袁美[4] 纪鸿飞[1,2,3] 李爱芹[2,3] 王兴军[1,2,3] 

机构地区：[1]山东省农业科学院高新技术研究中心/山东省作物与畜禽品种改良生物技术重点实验室,山东济南250100 [2]山东师范大学生命科学学院,山东济南250014 [3]农业部黄淮海作物遗传改良与生物技术重点开放实验室,山东济南250100 [4]山东省花生研究所,山东青岛266100

出　　处：《核农学报》2012年第1期43-48,共6页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金(31000720);山东省优秀中青年科学家奖励基金(2006BS06008);山东省自然科学基金(Y2008D44;ZR2010CZ002);山东省泰山学者岗位基金

摘　　要：通过构建花生未成熟种子cDNA文库,结合大规模EST测序,首次从花生中克隆了原花青素代谢途径上的2个关键酶[肉桂酸-4-羟化酶(C4H)和花青素还原酶(ANR)]基因的全长编码序列。序列分析表明,与其他植物的C4H相比,花生C4H整个编码区高度保守,开放阅读框长度1518bp,编码蛋白长度505个氨基酸,预测的蛋白分子量为57.9kDa,等电点为9.04。花生中ANR开放阅读框长度966bp,编码蛋白长度321个氨基酸,预测的蛋白分子量为35kDa,等电点为8.31。亚细胞定位预测分析表明,C4H定位在线粒体中,ANR则定位在胞外。半定量RT-PCR结果表明,C4H在花生根、茎、叶、花、果针、种子中均有表达,但在茎、叶中表达相对较低,而原花青素合成途径中的关键酶ANR在各个组织中均有表达,其中在果针中表达最强。By constructing a cDNA library of peanut immature seed and large scale EST sequencing, full length open reading frame of peanut cinnamate 4-hydroxylase （C4H） and anthocyanidin reductase （ANR） were cloned. Sequence alignment results showed that the whole coding region of C4H was highly conserved among different plant species. The ORF of AhC4H was 1518bp, which encoded a protein of 505 amino acids with a predicted molecular weight of 57.9kDa and an isoelectric point of 9.04. AhANR with an ORF of 966bp encoded a protein of 321 amino acids which had a predicted molecular weight of 35kDa and an isoelectric point of 8.31. Subeellular localization analysis indicated that AhC4H was located in mitochondria and AhANR in extracellular. Semi-quantitative PCR results demonstrated that AhC4H and AhANR were ubiquitously expressed in root, stem, leaf, flower, gynophore and seed. The expression of AhANR was strong and higher in gynorphore than in other tissues.

关 键 词：花生 CDNA文库 基因克隆 基因表达 

分 类 号：S565.2[农业科学—作物学]