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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。
The full length cDNA of flavanone 3-hydroxylase (F3H) gene was cloned by RACE method using the fruits of Ribes rubrum L. and R.albrum L., Named Rr F3H and Ra F3H respectively (KU984435 and KU984436). The Rf F3H was 1 351 bp in length and 1 098 bp in open reading frame encoding 365 amino acids. The full length of Rf F3H was 1 291 bp with an open reading frame of 1 098 bp encoding a protein of 365 amino acids. The amino acid sequence alignment showed that the protein encoded by this gene has DIOX-N superfamily and typical F3H protein functional domain (2OG-FeⅡ-Oxy oxygenase domain) Enzyme superfamily Phylogenetic analysis showed that Rr F3H and Ra F3H have obvious species characteristics in evolution and belong to relatively independent evolutionary branch. Quantitative PCR analysis showed that the expression level of F3H in red currant was much higher than that in white currant. In red currant, the expression level of F3H increased gradually with the coloring of fruit, and reached the highest level when the color of fruit was about 75% Decreased; the expression of the gene decreased with the growth of fruit in white currant, and the content of anthocyanin also showed a downward trend, indicating that F3H gene plays a role in the coloring of currant fruit.