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园艺学报 ›› 2019, Vol. 46 ›› Issue (3): 473-485.doi: 10.16420/j.issn.0513-353x.2018-0572

• 研究论文 • 上一篇    下一篇

苋菜中甜菜红素代谢相关基因AmMYB2的表达与功能分析

彭丽云,王 云,孙雪丽,王 晓,赵春丽,王丛巧,项蕾蕾,陈家兰,赖钟雄*,刘生财*   

  1. (福建农林大学园艺植物生物工程研究所,福州350002)
  • 出版日期:2019-03-25 发布日期:2019-03-25

Expression and Functional Analysis of AmMYB2 Related to Betalain Metabolism of Amaranthus tricolor

PENG Liyun,WANG Yun,SUN Xueli,WANG Xiao,ZHAO Chunli,WANG Congqiao,XIANG Leilei,CHEN Jialan,LAI Zhongxiong*,and LIU Shengcai*   

  1. (Institute of Horticultural Plant Biological Engineering,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
  • Online:2019-03-25 Published:2019-03-25

摘要:

为了研究AmMYB2基因在苋菜(Amaranthus tricolor)甜菜红素合成中的作用,对红色苋菜品种‘大红’AmMYB2(登录号为KY814751.1)进行生物信息、基因表达及功能分析。结果表明:AmMYB2的开放阅读框(ORF)全长为678 bp,编码蛋白分子质量为25.96 kD,该蛋白具有R2R3型MYB结构域,属于R2R3型MYB转录因子。核酸序列分析结果显示,AmMYB2与甜菜红素合成相关的甜菜BvMYB1和苋菜AmMYB1的相似度分别达到62.11%和49.93%。亚细胞观察显示,AmMYB2蛋白定位于细胞核。实时荧光定量(qPCR)分析发现,AmMYB2在红色苋菜品种的表达量高于绿色苋菜品种;在‘大红’苋菜中,不同组织以及叶片不同部位中AmMYB2的表达量与甜菜红素含量成正比。构建AmMYB2过表达载体,将其转入烟草和拟南芥中,发现该基因能够在这两种植物中高水平表达,但不会引起颜色变化,揭示AmMYB2可能是苋菜中甜菜红素合成的正调控因子,可能不参与花青素的合成。

关键词: 苋菜, AmMYB2基因, 甜菜红素, 表达分析, 功能鉴定

Abstract:

In order to explore the role of AmMYB2 in the betalain synthesis in Amaranthus tricolor,the bioinformatics,gene expression and functional analysis of AmMYB2(Genbank accession No. KY814751.1)in the red amaranth cultivar‘Dahong’were conducted. The results showed that the open reading frame(ORF)of the AmMYB2 is 678 bp in length. The coding protein molecular mass fraction is 25.96 kD,which belongs to R2R3 MYB transcription factor with R2R3 MYB domain. Subcellular localization assay revealed that the AmMYB2 protein was localized in the nucleus. Nucleotide sequence analysis revealed that the AmMYB2 is relatively close to betalain synthesis-related BvMYB1 and AmMYB1,with an identity of 62.11% and 49.93%,respectively. Quantitative real-time PCR(qRT-PCR)results showed that the expression level of AmMYB2 was higher in red amaranth cultivar than that in green,and was different in different tissues and the different parts of leaves from the‘Dahong’amaranth,consistent with the proportional to the content of betalains. After the over expression vector of AmMYB2 was constructed and transferred to tobacco and Arabidopsis thaliana,the expression level of AmMYB2 was higher in both plants,without color changes. Therefore,the AmMYB2 may be a positive regulatory factor for the synthesis of betalains in amaranth,and maybe not involved in anthocyanin synthesis.

Key words: Amaranthus tricolor, AmMYB2, betalain, expression analysis, functional identification

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