金荞麦C2H2-ZFP家族基因分析及FdC2H2-2在芦丁合成积累中的功能表征  

Analysis of C2H2-ZFP Family Genes and Functional Characterization of FdC2H2-2 in Rutin Synthesis and Accumulation in Golden Buckwheat

作　　者：龙欧 卢翔[1,2] 石亚亮 李光胜 张凯旋 王俊珍 周美亮 LONG Ou;LU Xiang;SHI Ya-liang;LI Guang-sheng;ZHANG Kai-xuan;WANG Jun-zhen;ZHOU Mei-liang(Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 100081;College of Agriculture,Guizhou University,Guiyang 550025;College of Life and Health,Hunan University of Science and Technology,Xiangtan 411100;Agricultural Science Research Institute of Liangshan Yi Autonomous Prefecture,Xichang 615000,Sichuan)

机构地区：[1]中国农业科学院作物科学研究所,北京100081 [2]贵州大学农学院,贵阳550025 [3]湖南科技大学生命与健康学院,湘潭411100 [4]凉山彝族自治州农业科学研究院,四川西昌615000

出　　处：《植物遗传资源学报》2023年第4期1161-1173,共13页Journal of Plant Genetic Resources

基　　金：国家重点研发计划(2019YFD1001300,2019YFD1001304);国家自然科学基金国际(地区)合作与交流项目(32111540258);湖南省科技厅湖湘高层次人才聚集工程项目(2020RC5031)。

摘　　要：C2H2锌指蛋白转录因子在植物生长发育、胁迫响应和次生代谢合成调控等方面发挥重要作用。前期研究发现金荞麦根部类黄酮含量高于苦荞麦可能是由于FdCHI,FdF3H,FdDFR等与类黄酮生物合成相关的基因家族被扩增。然而,参与类黄酮生物合成的C2H2锌指蛋白转录因子家族基因在金荞麦中如何调控芦丁合成尚未见报道。本研究对金荞麦FdC2H2-ZFP转录因子进行了全基因组鉴定和表达谱分析。共鉴定出114个FdC2H2-ZFPs。在对RNA-Seq数据分析基础上,筛选并克隆C2H2锌指蛋白基因FdC2H2-2。该基因具有3个典型的C2H2锌指结构,与拟南芥AtTREE1、AtDAZ3同源性较高。qRT-PCR显示FdC2H2-2基因的表达显著受茉莉酸诱导。此外,过表达FdC2H2-2毛状根中芦丁含量显著高于对照,且其芦丁合成途径中关键酶基因黄酮醇合成酶(FLS)、苯丙氨酸解氨酶(PAL)与类黄酮3’,5’-羟化酶(F3’5’H)表达量显著提高。以上结果表明,金荞麦FdC2H2-2基因可能通过激活芦丁生物合成关键酶基因FLS、PAL以及F3’5’H的表达,从而正向调控芦丁的积累。本研究为今后解析金荞麦C2H2锌指蛋白基因的功能提供参考。C2H2 zinc finger protein transcription factor plays an important role in plant growth and development,stress response and regulation of secondary metabolism synthesis.The previous study found that the flavonoid content in the root of fagopybuckwheat was higher than that of Tartary buckwheat,which may be due to the amplification of gene families related to flavonoid biosynthesis,such as FdCHI,FdF3H and FdDFR.However,it has not been reported how the C2H2 zinc finger protein transcription factor family genes involved in flavonoid biosynthesis regulate rutin synthesis in golden buckwheat.In this study,the whole genome of FdC2H2-ZFP transcription factor was identified and its expression profile was analyzed.A total of 114 FdC2H2-ZFPs were identified.Based on the analysis of RNA-Seq data,the C2H2 zinc finger protein gene FdC2H2-2 was screened and cloned.This gene has three typical C2H2 zinc finger structures,and has high homology with Arabidopsis AtTREE1 and AtDAZ3.qRT-PCR showed that the expression of FdC2H2-2 gene was significantly induced by jasmonic acid.In addition,the content of rutin in the hairy roots of overexpressing FdC2H2-2 was significantly higher than that of the control,and the transcriptional expression of flavonol synthetase(FLS),phenylalanine aminolyase(PAL)and flavonoid 3',5'-hydroxylase(F3'5'H),the key enzyme genes in the rutin synthesis pathway were significantly increased.These results suggest that FdC2H2-2 gene may positively regulate the accumulation of rutin by activating the expressions of the key enzymes in rutin biosynthesis,such as FLS,PAL and F3'5'H.This study provides reference for the future analysis of C2H2 zinc finger protein gene function of golden buckwheat.

关 键 词：金荞麦 C2H2锌指蛋白 全基因组鉴定 系统进化 基因克隆 芦丁合成 

分 类 号：S517[农业科学—作物学]