桑树花青素还原酶基因MaANR的克隆和表达分析  

Cloning and Expression Analysis of Anthocyanidin Reductase Gene from Mulberry

作　　者：李军[1] 梁燕梅[2] 赵爱春[2] 刘长英[2] 吕蕊花[2] 刘晓清[1] 余茂德[2] 

机构地区：[1]贵阳中医学院,贵阳550025 [2]西南大学生物技术学院,重庆400715

出　　处：《蚕业科学》2016年第4期570-575,共6页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(No.31360190);贵州省社会发展攻关项目(No.黔科合SY[2015]3030)

摘　　要：花青素还原酶(ANR)是原花青素单体生物合成过程中的关键酶之一,对花青素在植物组织中的积累有重要调控作用。通过检索川桑(Morus notabilis)基因组数据库鉴定获得了ANR候选基因序列,并以人工四倍体果桑品种嘉陵30号的桑椹c DNA为模板克隆了ANR基因(Ma ANR),其CDS序列长1 014 bp,编码337个氨基酸残基。聚类分析结果表明Ma ANR与草莓、西洋梨的花青素还原酶Fa ANR和Pc ANR的亲缘关系较近。实时荧光定量PCR检测Ma ANR在桑椹中的表达量最高,在桑树其他部位和组织中的表达量极低,在茎中几乎检测不到表达;Ma ANR在生长发育早期的桑椹中表达量较低,在快速生长过程的桑椹中高量表达,而在花青素大量合成的桑椹成熟时期表达量极低。初步推测Ma ANR可能是桑椹中花青素积累的重要负调控因子。Anthocyanidin reductase（ ANR） is one of the key enzymes in biosynthesis of proantho cyanidins,which has an important regulatory role in anthocyanin accumulation in plant tissues. In this study,a candidate ANR gene was identified from Morus notabilis genome database. Ma ANR gene was cloned from Jialing 30,a tetraploid mulberry variety for fruit use,using mulberry fruit c DNA as template. The amplified coding sequence of Ma ANR is 1 014 bp,encoding 337 amino acids. Cluster analysis result indicated that Ma ANR had closer relationship with Fa ANR from Fragaria ananassa and Pc ANR from Pyrus communis. Real time fluorescent quantitative PCR results indicated that the expression level of Ma ANR was the highest in mulberry fruits,was very low in other positions and tissues,and was scarcely expressed in stem. The expression level of Ma ANR was low in mulberry fruits at the early developmental stage,was high at the fast growing stage,and was the lowest at the mature stage. It was preliminarily inferred that Ma ANR may be an important negative regulator in anthocyanins accumulation.

关 键 词：桑树 花青素还原酶 基因克隆 表达模式 桑椹 发育时期 

分 类 号：S888.2[农业科学—特种经济动物饲养] Q943.2[农业科学—畜牧兽医]