紫色番茄黄烷酮3-羟化酶基因SlF3H-like克隆与表达分析  

Cloning and Expression Analysis of Flavanone 3-hydroxylase SlF3H-like Gene From Purple Tomato

作　　者：刘正杰[1,2] 刘运通 徐绍忠[1] 毛自朝[1,2] 林春[1,2] LIU Zhengjie;LIU Yuntong;XU Shaozhong;MAO Zichao;LIN Chun(College of Agriculture and Biotechnology,Yunnan Agricultural University,Kunming,Yunnan 650201;Institute of Improvement and Utilization of Characteristic Resource Plants,Yunnan Agricultural University,Kunming,Yunnan 650201)

机构地区：[1]云南农业大学农学与生物技术学院,云南昆明650201 [2]云南农业大学特色小宗作物研究中心,云南昆明650201

出　　处：《北方园艺》2022年第17期26-33,共8页Northern Horticulture

基　　金：云南省农业基础研究联合专项面上资助项目(2018FG001(-019));云南省基础研究计划面上资助项目(202101AT070714)。

摘　　要：以樱桃型紫色番茄‘砚紫1号’为试材,采用RT-PCR、生物信息学与qRT-PCR表达分析的方法,研究了该材料中黄烷酮3-羟化酶基因SlF3H-like的生物学信息,以期表征该基因序列特征和表达模式。结果表明:SlF3H-like基因开放阅读框(ORF)长度为1 089 bp,编码362个氨基酸;随机卷曲占据SlF3H-like蛋白结构元件的大多数,其次是α-螺旋和延伸链;‘砚紫1号’与其它物种的F3H氨基酸序列保守性非常高,SlF3H-like蛋白含有典型的非血红素双加氧酶和2OG-FeⅡ_Oxy加氧酶结构域;系统进化分析发现‘砚紫1号’与‘潘那利’番茄亲缘关系最近,其次是马铃薯。测定‘砚紫1号’发育过程中叶片和果皮中花青素含量,发现果皮中花青素含量随着果实直径的变大而逐渐积累,而叶片中花青素含量随着叶片直径的变大表现为先升后急剧下降。qRT-PCR分析SlF3H-like基因在‘砚紫1号’发育过程中果皮和叶片内的表达,发现该基因表达变化趋势与花青素含量的变化趋势类似,说明SlF3H-like基因在紫色番茄花青素积累的过程中起到重要的作用。The cherry purple tomato ‘Yanzi No.1’ was used as test material.RT-PCR,bioinformatics and qRT-PCR expression analysis were used to study the biological information of flavanone 3-hydroxylase gene SlF3 H-like,in order to characterize the sequence characteristics and expression pattern of the gene.The results showed that the open reading frame(ORF) of SlF3H-like gene was 1 089 bp, encoding 362 amino acids.The random coil was found to occupy the majority of structural elements of SlF3 H-like protein, followed by α-helix and extended chain.The F3 H amino acid sequence of ‘Yanzi No.1’ was highly conserved from other species.SlF3 H-like protein contained typical DIOX-N superfamily and 2 OG-FeⅡ_Oxy domains.Phylogenetic analysis showed that ‘Yanzi No.1’ was closely related to ‘Pannali’ tomato, followed by potato.The anthocyanin content in leaves and pericarp of ‘Yanzi No.1’ was measured during its development.It was found that the anthocyanin content in pericarp gradually accumulated with the increase of fruit diameter, while the anthocyanin content in leaves increased first and then decreased sharply with the increase of leaf diameter.qRT-PCR was used to analyze the expression of SlF3 H-like gene in the pericarp and leaves of ‘Yanzi No.1’.Results showed that the change trend of this gene expression was similar to that of anthocyanin content, indicating that SlF3 H-like gene played an important role in anthocyanin accumulation of purple tomato.

关 键 词：紫色番茄 SlF3H-like基因 克隆 花青素 表达分析 

分 类 号：S641.2[农业科学—蔬菜学]