作物学报2018，Vol.44Issue(7)：1021-1031,11.DOI:10.3724/SP.J.1006.2018.01021
马铃薯块茎花色素苷合成相关R2R3 MYB蛋白基因的克隆和功能分析
Cloning and Functional Analysis of R2R3 MYB Genes Involved in Anthocyanin Biosynthesis in Potato Tuber
摘要
Abstract
The anthocyanins biosynthesis is regulated by transcription factors, and R2R3 MYB is the most important transcrip-tional regulator in plant. In this study, three homologous genes were isolated from tetraploid potato, belonging to R2R3 MYB gene family. The structure and function analysis of the three homologous genes were characterized by bioinformatics analysis, stable tobacco genetic transformation and qPCR assays. The three homologous genes contained R2 and R3 conserved domains, differed in the number of repeats (R) consisting of 10 amino acid sequences, and named as StAN1-R0, StAN1-R1, and StAN1-R3 according to the number of R. The coding proteins are hydrophilic with molecular weight of 28 047.91, 29 458.35, 31 527.60 Da, and isoelectric points (pI) of 6.14, 6.90, and 8.39, respectively. The accumulation of anthocyanin was significantly increased in StAN1-R0, StAN1-R1 and StAN1-R3 overexpressed plants. The leaf color of StAN1-R1-overexpressed plants was dark red with the highest anthocyanin content among the three transgenic events. The qPCR assays showed that exogenous StAN1 genes enhanced the expression of endogenous NtbHLH transcription factor as well as NtCHS, NtCHI, NtF3H, NtF3’H, NtDFR , NtANS, and NtUFGT, involved in anthocyanin biosynthesis in transgenic tobacco leaves. The overexpression of StAN1-R1 resulted in higher expression of NtDFR, NtANS, and endogenous NtbHLH in transgenic tobacco. The results showed that three homologous genes of StAN1 can regulate anthocyanin biosynthesis, among them StAN1-R1 containing one R has the strongest regulatory capacity.关键词
马铃薯/花色素苷/R2R3 MYB/基因克隆/生物信息学分析/烟草转化/基因表达分析Key words
potato/anthocyanin/R2R3 MYB/gene cloning/bioinformatics analysis/stable tobacco transformation/gene expres-sion analysis引用本文复制引用
谈欢,刘玉汇,李丽霞,王丽,李元铭,张俊莲..马铃薯块茎花色素苷合成相关R2R3 MYB蛋白基因的克隆和功能分析[J].作物学报,2018,44(7):1021-1031,11.基金项目
本研究由国家自然科学基金项目(31601356), 甘肃省杰出青年科学基金项目(17JR5RA138), 甘肃省农业生物技术研究与应用开发专项(GNSW-2015-15), 甘肃省高等学校科研项目(2016A-030)和甘肃农业大学"伏羲人才"计划项目(Gaufx-02Y04)资助.This study was supported by the National Natural Science Foundation of China (31601356), Gansu Science Foundation for Distinguished Young Scholars (17JR5RA138), the Agricultural Biotechnology Research and Application Development Foundation of Gansu Province (GNSW-2015-15), Gansu Scientific Research Foundation for the Universities (2016A-030), and Fuxi Talent Project of Gansu Agricultural University (Gaufx-02Y04). （31601356）
