红穗和白穗醋栗F3H的克隆及其在果实成熟过程中的表达分析  

Molecular Cloning and Expression Analysis of F3H Gene in Ribes rubrum and R.albrum During Fruit Mature

作　　者：王海竹[1,2] 闫海芳[2] 徐启江[1,2] WANG Hai-zhu YAN Hai-fang and XU Qi-jiang(The State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040, China College of Life Sciences, Northeast Forestry University, Harbin 150040, China)

机构地区：[1]东北林业大学林木遗传育种国家重点实验室,哈尔滨150040 [2]东北林业大学生命科学学院,哈尔滨150040

出　　处：《园艺学报》2016年第10期2003-2011,共9页Acta Horticulturae Sinica

基　　金：国家基础科学人才培养基金项目(J1210053);中央高校基本科研业务费专项资金项目(DL12CA10;2572014EA03-01);林木遗传育种国家重点实验室创新项目(2013A06;2013B010);黑龙江省博士后科研启动金项目(LBH-Q14011)

摘　　要：以红穗醋栗(Ribes rubrum L.)和白穗醋栗(R.albrum L.)果实为试材,采用RACE方法克隆黄烷酮3–羟化酶(F3H)基因c DNA全长序列,分别命名为Rr F3H和Ra F3H(KU984435和KU984436)。Rr F3H全长1 351 bp,开放阅读框长1 098 bp,编码365个氨基酸;Ra F3H全长1 291 bp,开放阅读框长1 098 bp,编码365个氨基酸。氨基酸多序列比对表明该基因编码的蛋白具有非血红素双加氧酶结构域(DIOX-N superfamily)和典型的F3H蛋白功能结构域(2OG-FeⅡ_Oxy加氧酶结构域),属于双加氧酶超家族。系统发育分析表明,Rr F3H和Ra F3H在进化上具有明显种属特性,属于相对独立的进化分支。定量PCR分析表明,F3H在红穗醋栗中表达量远远高于白穗醋栗,在红穗醋栗中随着果实着色加深表达量逐渐上升,果实着色约75%时表达量最高,之后下降;白穗醋栗中随着果实生长,该基因的表达下降,花色苷含量也呈下降趋势,说明F3H基因在醋栗果实着色过程中发挥作用。Red currant（Ribes rubrum L.）and white currant（R. albrum L.）fruits were used as materials,two full-length c DNA sequences of F3H gene were cloned using RACE（rapid amplification of c DNA ends）,and denoted as RrF3H and RaF3H. Both of them contained a 1 098 bp open reading frame that encoded a deduced protein with 365 amino acid residues. Amino acid sequence alignment showed that the protein had a non-heme dioxygenase domain（DIOX-N superfamily）and 2OG-FeⅡ-dependent dioxygenase superfamily. Phylogenetic analysis showed that,RrF3H and RaF3H had obvious characteristics of species in evolution,which had an relative independent evolutionary branch. Quantitative RT-PCR analysis showed that,during the period of fruit development F3 H expression in red currant was much higher than that in white currant. With the deepening of fruit colour,the expression level gradually increased until the period of 75% fruit colour,then the RrF3H decreased rapidly. With the fruit growth of white current,the expression pattern of F3H decreased. And the content of anthocyanins in the same period also showed a decreasing trend. All the results suggested that F3H play a role in the process of fruit coloring.

关 键 词：红穗醋栗 白穗醋栗 花色苷 F3H 基因克隆 

分 类 号：S663.9[农业科学—果树学]