茶树类黄酮3′-羟化酶基因的克隆与表达特性分析

发布时间：2018-03-15 00:08

本文选题：茶树　切入点：类黄酮′-羟化酶　出处：《茶叶科学》2017年01期 　论文类型：期刊论文

【摘要】：类黄酮3′-羟化酶(Flavonoid 3′-hydroxylase,F3′H)是细胞色素P450酶家族(Cytochrome P450,CYP450)的单加氧酶,在植物次生代谢和逆境调控方面起着重要的作用。本实验以茶树品种龙井43作为试验材料,利用RT-PCR方法,从c DNA中克隆得到茶树中编码类黄酮3′-羟化酶基因,命名为Cs F3′H1。序列分析显示,Cs F3′H1基因开放阅读框为1 530 bp,编码509个氨基酸,含有相对保守的P450酶系结合域。进化树分析表明,Cs F3′H1与Cs F3′H3的进化树关系相近,与Cs F3′H2的进化树关系较远。序列多重比对显示,Cs F3′H1蛋白具有F3′H蛋白的特征基序,并与高粱、猕猴桃、杨树、拟南芥和葡萄同源蛋白一致性为66.48%。氨基酸组分、理化性质、亲水性/疏水性和无序化分析显示,Cs F3′H1是亲水性蛋白,无序化特征不明显。利用实时荧光定量PCR对Cs F3′H1基因在茶树不同组织和不同激素处理下的表达谱进行检测,结果显示:不同组织中,Cs F3′H1基因的表达水平在第一叶中最高,之后随着叶片的成熟逐渐下降,Cs F3′H1基因在老叶和根中几乎不表达,表达水平从高到低依次为第一叶第二叶第三叶第四叶嫩茎根老叶;在不同激素处理中,Cs F3′H1在SA激素处理下的表达量最高,为对照的2.24倍,在Me JA处理下表达量最低,为对照的0.43倍。
[Abstract]:Flavonoid 3-hydroxylase (F3H) is a monooxygenase of cytochrome P450 (CYP450), which plays an important role in plant secondary metabolism and stress regulation. Longjing 43, a tea plant cultivar, was used as experimental material and RT-PCR method was used in this experiment. The coding gene of flavonoid 3-mo-hydroxylase in tea plant was cloned from c DNA and named as Cs F3H1. Sequence analysis showed that the open reading frame of CS-F3OH1 gene was 1 530bp, encoding 509 amino acids. The phylogenetic tree analysis shows that the phylogenetic tree of Cs F3H1 is similar to that of Cs F3H3, and the relationship is far from that of CS-F3H3.The sequence multiple alignment shows that the F3H1 protein has the characteristic motif of F3H1 and has the characteristic motif with sorghum. The homologous protein identity of kiwifruit, poplar, Arabidopsis thaliana and grape was 66.48.The amino acid composition, physicochemical properties, hydrophilicity / hydrophobicity and disordering showed that Cs F3H1 was a hydrophilic protein. The expression profiles of Cs F3OH1 gene in different tissues and different hormone treatments of tea plants were detected by real-time fluorescence quantitative PCR. The results showed that the expression level of Cs F3OH1 gene in different tissues was the highest in the first leaf. After that, the expression level of Cs F3H1 gene was almost not expressed in the old leaves and roots with the maturation of the leaves. The order of expression level from high to low was the first leaf, the second leaf, the third leaf, the third leaf, the tender stem root, and the old leaf, the first leaf, the second leaf, the third leaf and the young stem root. In different hormone treatments, the expression of Cs F3H1 was the highest under SA hormone treatment, 2.24 times of that of the control, and the lowest under me JA treatment, 0.43 times of the control.
【作者单位】：南京农业大学园艺学院茶叶科学研究所;
【基金】：国家自然科学基金(31570691)
【分类号】：S571.1

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