紫色马铃薯花青素StAN1基因的克隆及功能分析
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引用本文:贾羊毛加,王 芳,叶广继,王 舰.紫色马铃薯花青素StAN1基因的克隆及功能分析[J].西北植物学报,2019,39(3):397~403
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贾羊毛加1,王 芳1,2,3,4,5,叶广继1,2,3,4,5,王 舰1,2,3,4,5* (1 青海大学西宁 8100162 青海省农林科学院, 西宁 8100163 青海大学 青藏高原生物技术教育部重点实验室, 西宁 8100164 青海省马铃薯育种重点实验室,西宁 8100165 省部共建三江源生态与高原农牧业国家重点实验室, 西宁 810016) 
基金项目:国家自然科学基金(31660417);
中文摘要:MYB是植物花青素合成代谢途径中最主要的转录因子。该研究以紫色马铃薯B 5为试验材料,通过同源克隆技术,克隆了马铃薯花青素StAN1基因,并对StAN1基因的表达、遗传转化及其转基因烟草的花青素含量进行分析。结果表明:(1)StAN1基因全长774 bp,编码了258个氨基酸;系统进化树分析发现,StAN1与辣椒、茄子、芦竹的亲缘关系最近,与矮牵牛、苹果等的亲缘关系最远。(2)荧光定量PCR分析显示,StAN1基因在马铃薯不同组织均有表达,其表达量从小到大依次为:根<叶<茎<匍匐茎<薯皮<薯肉。(3)成功构建了表达载体pJAM1502 AN1,并经农杆菌(Gv3101)转化获得根、茎、叶片及叶脉均为紫红色的转StAN1基因烟草,PCR鉴定表明目的基因StAN1已成功转入烟草中。(4)花青素含量分析表明,野生型烟草叶片中花青素含量为2 mg/g,叶片颜色为绿色,而转StAN1基因烟草叶片花青素含量达20 mg/g,叶片颜色变成了紫红色。
中文关键词:紫色马铃薯  StAN1基因  克隆与转化  功能分析
 
Cloning and Function Analysis of Anthocyanin StAN1 Gene from Purple Potato
Abstract:MYB is the most important transcription factor in plant anthocyanin biosynthesis pathway. In this study, the anthocyanin StAN1 gene of purple potato was cloned by homologous cloning technique. The results showed that: (1) the full length of the StAN1 gene was 774 bp, encoded 258 amino acids; Phylogenetic tree analysis showed that StAN1 had the closest relationship with pepper, eggplant and asparagus, and had the furthest relationship with Petunia and apple. (2)Fluorescence quantitative PCR analysis showed that StAN1 gene was expressed in different tissues of potato. (3) The expression vector pJAM1502 AN1 was successfully constructed by Agrobacterium (Gv3101) transformation to obtain the root, stem, leaf and vein are purple red transgenic tobacco, the expression vector StAN1 gene was transformed from small to large as the root < leaf < stolon < tuber skin < tuber. (3) Transgenic tobacco with StAN1 gene was constructed successfully through Agrobacterium (Gv3101) transformation. PCR identification showed that the target gene StAN1 had been successfully transferred into tobacco. (4) The anthocyanin content of wild type tobacco leaves was 2 mg/g, and the color of leaves was green. However, the anthocyanin content of the transgenic tobacco leaves reached 20 mg/g, and the color of the leaves changed to purplish red.
keywords:purple potato  StAN1 gene  cloning and transformation  functional analysis
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