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2025年8月21日 星期四
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洋葱花青素合成相关基因(AcPAL1)的克隆和表达分析
梁毅1,刘小义2,张洪伟1,谭武平3
1. 北京市农林科学院蔬菜研究中心
2.
3. 北京市农林科学院蔬菜研究中心,农业部华北地区园艺作物生物学与种质创制重点实验室,北京京研益农科技发展中心
Cloning and Expression Analysis of an Anthocyanin Bio-synthesis-related Gene(AcPAL1) in Onion(Allium cepa L.)
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摘要 苯丙氨酸解氨酶(PAL)作为植物苯丙烷类代谢途径中的关键酶,在其生长发育、抗病抗逆等多种生命进程中起重要作用,是花青素生物合成途径中的第一个酶。为了研究洋葱(Allium cepa L.) PAL基因的生物学功能及其与花青素合成之间的关系,利用不同植物PAL基因设计简并引物,通过RT-PCR和RACE技术克隆洋葱PAL基因全长cDNA序列(GenBank登录号: KF421110),并对该基因进行序列分析和Real-time PCR表达分析。结果表明,该序列全长2 363 bp,编码包含708个氨基酸残基的蛋白质多肽;Blast分析和系统进化分析表明,该多肽与大蒜(Allium. sativum)、石蒜(Lycoris radiate)PAL蛋白相似性很高,因此被命名为AcPAL1。Real-time PCR表达分析结果表明,AcPAL1基因在白皮、黄皮和红皮洋葱中表达量依次增加;而随着鳞茎的不断膨大,其表达量却不断降低。本实验初步证实了所克隆的洋葱AcPAL1基因与花青素合成相关联,为研究洋葱PAL基因同花青素合成积累之间的关系提供了依据。
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梁毅
刘小义
张洪伟
谭武平
关键词 洋葱花青素AcPAL1    
Abstract:As a key enzyme of the plant phenylpropanoid pathway, phenylalanine ammonia-lyase (PAL), which plays an important role in the life processes such as growth and development, and disease resistance, is the first enzyme in the anthocyanin synthesis pathway. In order to study the biological function for PAL in onion(Allium cepa L.) and its relationship with anthocyanin synthesis, the full length cDNA of PAL gene(GenBank accession no. KF421110) in onion bulb was cloned by RT-PCR and RACE technology using degenerate primers designed according to PAL genes in different plants. The sequence analysis and expression analysis using Real-time PCR were performed either. The results showed that the cloned sequence designated AcPAL1 had full length of 2 363 bp, which encoded a protein polypeptide of 708 amino acids. Blast and phylogenetic analysis indicated that the polypeptide shared a very high similarity with Allium sativum and Lycoris radiate. Expression analysis using Real-time PCR demonstrated that the transcription of AcPAL1 increase by degrees in white, yellow and red onions, and the expression was down-regulated by the continuous expansion of the bulb. In this study it is demonstrated that the expression of AcPAL1 is somewhat related to the onion anthocyanin biosynthesis and provided the basis for their relationship study between the onion PAL gene and accumulation of anthocyanin synthesis.
Key wordsOnion    Anthocyanin    AcPAL1
收稿日期: 2013-07-23     
通讯作者: 张洪伟   
引用本文:   
梁毅1,刘小义2,张洪伟1,谭武平3. 洋葱花青素合成相关基因(AcPAL1)的克隆和表达分析[J]. , 2014, 22(1): 47-54.
, , ,. Cloning and Expression Analysis of an Anthocyanin Bio-synthesis-related Gene(AcPAL1) in Onion(Allium cepa L.). , 2014, 22(1): 47-54.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2014/V22/I1/47
 
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