葡萄风信子MaMYB114基因克隆及功能研究  

Cloning and Function Analysis of MaMYB114 Gene in Grape Hyacinth

作　　者：史程程 陈鹏[1] 焦清正 刘艺平[1] 刘红利 SHI Chengcheng;CHEN Peng;JIAO Qingzheng;LIU Yiping;LIU Hongli(College of Landscape Architecture and Art,Henan Agricultural University,Zhengzhou 450000,China)

机构地区：[1]河南农业大学风景园林与艺术学院,郑州450000

出　　处：《西北植物学报》2023年第12期2039-2048,共10页Acta Botanica Boreali-Occidentalia Sinica

基　　金：河南省科技攻关项目(222102110263);河南省高校国家级大学生创新创业训练计划项目(202110466004)。

摘　　要：MYB转录因子在植物花青素生物合成过程中发挥主要调控作用。为探究MYB转录因子在葡萄风信子(Muscari spp.)蓝色花色形成中的作用,研究以葡萄风信子‘亚美尼亚’为试验材料,基于课题组前期转录组数据库深度挖掘,经本地Blast比对分析,采用RT-PCR方法克隆获得1个R2R3-MYB基因,命名为MaMYB114(GenBank登录号:OQ615377),对其进行生物信息学分析和异源转化烟草功能验证。结果表明,(1)MaMYB114开放阅读框全长为714 bp,编码237个氨基酸;MaMYB114蛋白含有R2R3保守结构域,属于R2R3-MYB家族AN2亚家族。(2)亚细胞定位及转录自激活结果显示基因定位于细胞核,且具有转录自激活活性,符合转录因子一般特征。(3)实时荧光定量分析显示MaMYB114基因主要在花中高表达,表达模式具有组织特异性。结合不同时期花青苷含量变化模式,发现MaMYB114在着色期花青素含量高的花蕾中高表达,表明MaMYB114可能参与了葡萄风信子花青苷合成过程。(4)过表达MaMYB114促进烟草花青苷积累,表明MaMYB114正向调控花青素合成。本研究表明,MaMYB114基因可能是参与葡萄风信子蓝色花色形成的重要调控基因。MYB transcription factors play a major regulatory role in plant anthocyanin biosynthesis.In order to explore the role of MYB transcription factor in the formation of the blue flower color of Muscari armeniacum,this study used M.armeniacum as the experimental material.Based on the local deep mining of the transcriptome database of the research group,authors cloned one R2R3-MYB gene named MaMYB114(GenBank accession number:OQ615377)for bioinformatics analysis and functional validation of heterologous transformed tobacco.The results showed that,(1)the open reading frame of MaMYB114 had a total length of 714 bp,encoding a protein of 237 amino acids.Sequence alignment showed MaMYB114 protein contains the R2R3 conserved domain,indicating that it belongs to the AN2 subfamily of the R2R3-MYB family.(2)The results of subcellular localization and transcriptional self-activation confirmed that MaMYB114 was located in the nucleus and had transcriptional autoactivation activity,which was consistent with the general characteristics of transcription factors.(3)Real-time quantitative PCR analysis showed that MaMYB114 gene was highly expressed mainly in flowers,and the expression pattern was tissue-specific.Combined with the change patterns of anthocyanin content at different times,it was found that MaMYB114 was highly expressed in flower buds with high anthocyanin content during the coloring stage,indicating that MaMYB114 participated in the synthesis process of anthocyanins in grape hya-cinths.(4)Overexpression of MaMYB114 promotes anthocyanins accumulation in tobacco,indicating that MaMYB114 positively regulates anthocyanin synthesis.This study indicated that the MaMYB114 gene might be an important regulatory gene involved in the formation of the blue color of grape hyacinth.

关 键 词：葡萄风信子 花青素苷 MYB转录因子 异源过表达 基因功能 

分 类 号：Q785[生物学—分子生物学] Q786Q789