‘胎里红’枣果皮花青素合成酶ZjANS基因克隆及表达分析  

Cloning and Expression Analysis of Anthocyanidin Synthase Gene ZjA NS from the Peel of Ziziphus jujuba cv.'Tailihong'

作　　者：祝怛均 石倩倩 李希 杜江涛 李新岗[1] Zhu Dajun;Shi Qianqian;Li Xi;Du Jiangtao;Li Xingang(Shaanxi Forestry Comprehensive Laboratory,College of Forestry,Northwest A&F University,Yangling,712100)

机构地区：[1]西北农林科技大学林学院,陕西省林业综合实验室,杨凌712100

出　　处：《分子植物育种》2023年第20期6613-6624,共12页Molecular Plant Breeding

基　　金：国家重点研发计划项目(2022YFD2200404);陕西省科技创新课题(2021XYF-11)共同资助。

摘　　要：为了研究枣果实发育过程中花青苷的积累和相关基因的表达模式,以紫色枣品种‘胎里红’为研究材料,利用枣基因组数据克隆获得两条ANS基因全长序列,将其命名为ZjANS1和ZjANS2。其开放阅读框分别为888和648 bp,分别编码295和215个氨基酸。蛋白多序列比对和进化树分析表明,ZjANS1和ZjANS2具有典型的2OG-Fe(II)-Oxy保守结构域。蛋白互作预测发现,含有LDOX/ANS结构域的蛋白主要与MYB113及DFR、UF3GT、F3H、TT8等存在互作关系。ZjANS1和ZjANS2与甜橙CsANS蛋白序列聚为一支,亲缘关系较近。亚细胞定位分析发现,ZjANS1和ZjANS2的功能结构域均定位在内质网上。RT-qPCR结果表明,ZjANS1和ZjANS2在不同组织中均有表达,但表达存在差异。ZjANS1在茎、芽和幼果中表达量较高,在成熟叶中最低,而ZjANS2在茎中表达量相对较高,在其他组织中较低。花青苷含量测定结果表明‘,胎里红’果实发育前期总花青苷和各花青苷组分的含量较高,后期花青苷含量较少。此外,将花青苷合成相关结构基因表达模式和与总花青苷含量进行关联分析,结果表明ZjDFR、ZjANS1和ZjUFGT表达模式类似,并且与花青苷含量呈极显著正相关。由此推测ZjANS1、ZjDFR和ZjUFGT是‘胎里红’果皮花青苷合成的关键基因,对‘胎里红’果皮色素合成及颜色转变中起着重要的作用。本研究可为深入研究枣果实花青苷合成机理提供理论基础。In order to study the accumulation of anthocyanins and the expression patterns of anthocyanin-related genes during the development of jujube fruit,the purple jujube variety'Tailihong'was used as the research material to clone two full-length sequences of A NS genes named ZjA NSI and ZjANS2 basing on the jujube genome data.Their open reading frames are 888 and 648 bp,encoding 295 and 215 amino acids,respectively.Protein sequence alignment and phylogenetic tree analysis showed that ZjANS1 and ZjANS2 have typical 2OG-Fe(II)-Oxy conserved domains.Protein interaction prediction found that LDOX/ANS domain containing proteins mainly interacted with MYB113 and DFR,UF3GT,F3H,TT8,etc.ZjANS1 and ZjANS2 are closely related to the sweet orange CsANS protein.Subcellular localization analysis found that the functional domains of ZjA NSI and ZjA NS2 are located in the endoplasmic reticulum.The results of RT-qPCR showed that ZjANSI and ZjANS2 are expressed in different tissues,but the expression is different.ZjANSI has a higher expression level in stems,buds and young fruits,and is the lowest in matured leaves,while ZjANS2 has a relatively higher expression level in stems and low-er levels in other tissues.The results of anthocyanin content determination showed that the total anthocyanin contents and the contents of each anthocyanin component in the early ripening stage of'Tailihong'fruit are higher,and the anthocyanin content is lower in the later stage.In addition,the correlation analysis between the expression patterns of functional genes related to anthocyanin synthesis and total anthocyanin contents showed that the expression patterns of ZjDFR,ZjANSI and ZjUFGT were similar,and had a very significant positive correlation with the anthocyanin content.It is inferred that ZjA NS1,ZjDFR and ZjUFGT are the key genes responsible for the anthocyanin synthesis of'Tailihong'peel,and play an important role in pigment synthesis and coloring of'Tailihong'pericarp.This study may provide a theoretical basis for in-depth study of the mecha

关 键 词：枣(Ziziphus jujuba) ANS 花青苷 基因克隆 表达分析 

分 类 号：S665.1[农业科学—果树学]