甜荞花青素合成相关基因FeR2R3-MYB的克隆与表达分析

Cloning and expression analysis of FeR2R3-MYB of anthocyanin synthesis-related genes of common buckwheat

关键词:  甜荞, MYB 转录因子, 生物信息学, 亚细胞定位, 表达分析

Key words:  common buckwheat, MYB transcription factor, bioinformatics, subcellular location, expression analysis

摘要:

MYB 是一类常见的转录因子,广泛参与植物花青素生物合成的调控。为探究 MYB转录因子在甜荞花青素生物合成中的调控作用,该研究从红花甜荞和白花甜荞转录组学数据中筛选并克隆出一个和花青素生物合成相关的MYB基因,将其命名为 FeR2R3-MYB,GenBank 登录号为 MT151381.1,并对该序列进行生物信息学分析,以及利用 qRT-PCR 分析FeR2R3-MYB基因在白花甜荞和红花甜荞中的表达特征。结果表明:(1)FeR2R3-MYB基因全长 831 bp,编码 276 个氨基酸,蛋白的相对分子质量为 30.95 kD,理论等电点(pI)为 8.73,蛋白的不稳定指数为 69.64,属于不稳定蛋白,总疏水值为-0.679,整条肽链呈现亲水特性。(2)FeR2R3-MYB 具有典型的 R2R3-MYB 结构域,属于 R2R3-MYB 亚家族。(3)FeR2R3-MYB 与同属蓼科的苦荞和虎杖亲缘关系比较近。(4)FeR2R3-MYB 的启动子序列共含有 9 个光照响应元件、17 个转录因子结合位点、4 个非生物响应元件和 2 个激素响应元件。(5)亚细胞定位发现 FeR2R3-MYB 只在细胞核中表达。(6)FeR2R3-MYB 基因的表达量在叶片和花序中红花甜荞均高于白花甜荞,推测 FeR2R3-MYB 基因可以正向调节甜荞花青素生物合成。综上所述,该研究结果为进一步深化 FeR2R3-MYB 基因在甜荞花青素生物合成途径中的功能及表达调控方面的研究提供了基础。

Abstract:

MYB is a common transcription factors widely involved in the regulation of anthocyanidin biosynthesis. In order to explore the regulatory role of MYB transcription factors in the biosynthesis of common buckwheat anthocyanidins, this study screened and cloned a MYB gene associated with anthocyanin biosynthesis from the transcriptomic data of safflower common buckwheat and white flower common buckwheat, and named it FeR2R3-MYB, GenBank login number was MT151381.1. The sequence was analyzed by bioinformatics analysis and qRT-PCR was used to analyze the expression characteristics of FeR2R3-MYB gene in white flower common buckwheat and safflower common buck wheat. The results were as follows:(1)FeR2R3-MYB gene was 831 bp in total length, encoding 276 amino acids. The relative molecular mass of the protein was 30.95 kD, the theoretical isoelectric point(pI)was 8.73, and the instability index of the protein was 69.64, which belonged to the unstable protein. The total hydrophobic value was -0.679, and the whole peptide chain showed hydrophilic characteristics.(2)FeR2R3-MYB had a typical R2R3-MYB domain and belonged to the R2R3-MYB subfamily.(3)FeR2R3-MYB was closely related to common buckwheat and knotweed, belonging to the same family.(4)The promoter sequence of FeR2R3-MYB contained a total of nine light corresponding elements, 17 transcription factor binding sites, four abiotic corresponding elements and two hormone response elements.(5)Subcellular localization found that FeR2R3-MYB was only expressed in the nucleus.(6)The expression of FeR2R3-MYB gene of safflower common buckwheat was higher than that of white flower common buckwheat in leaves and inflorescences, and it was further speculated that FeR2R3-MYB gene could positively regulate the biosynthesis of common buckwheat anthocyanin. In summary, these results lay a foundation for further deepening the research on the function and expression regulation of FeR2R3-MYB gene in the biosynthetic pathway of common buckwheat anthocyanin.