滇牡丹MYB家族成员鉴定及PdMYB2的功能验证  

Identification of MYB Family Members and Functional Verification of PdMYB2 in Paeonia delavayi

作　　者：邹红竹 韩璐璐 周琳[1] 吕纪杭 贾莹华 王雁[1] ZOU Hong-zhu;HAN Lu-lu;ZHOU Lin;LV Ji-hang;JIA Ying-hua;WANG Yan(State Forestry Administration,Key Laboratory of Tree Breeding and Cultivation,Research Institute of Forestry,Chinese Academy of Forestry,Beijing 100091,China)

机构地区：[1]中国林业科学研究院林业研究所,国家林业和草原局林木培育重点实验室,北京100091

出　　处：《林业科学研究》2022年第5期1-13,共13页Forest Research

基　　金：国家重点研发计划项目(2018YFD1000405);国家自然科学基金项目(31972456)。

摘　　要：[目的]研究MYB转录因子家族在滇牡丹黄色花花色变黄过程中的重要作用,为牡丹花色改良和分子育种提供候选基因。[方法]本研究基于滇牡丹花瓣的转录组数据,对滇牡丹MYB家族进行了成员鉴定及分类。依据系统进化树、同源序列比对、基因差异表达筛选到候选基因PdMYB2,利用亚细胞定位、实时荧光定量PCR、转基因烟草及液相色谱等方法对PdMYB2进行功能验证。[结果]在滇牡丹转录组中共鉴定到64个MYB基因家族成员,分别为R2R3-MYB成员25个,R1R2R3-MYB 1个,4R-MYB 2个,MYB-related 36个。筛选到1个R2R3-MYB转录因子PdMYB2,定位在细胞核和细胞质中,在烟草中过表达PdMYB2会引起烟草花青素积累量的增加,在花瓣中与花青素相关的结构基因NtCHS和NtCHI的表达量均上调。[结论]MYB转录因子家族中的Pd MYB2在类黄酮合成途径中具有正调控作用,可以促进花青素的积累,提高类黄酮途径上结构基因的表达量。[Objective]The role of MYB transcription factor family in pigment formation of yellow Paeonia delavayi flowers was analyzed to provide candidate genes for color improvement and molecular breeding of tree peony.[Method]In this study,members of MYB gene family of P.delavayi were identified and classified based on the transcriptome data of petals.One member of R2R3-MYBs,PdMYB2,was selected as a candidate gene according to phylogenetic tree,homologous sequence alignment and gene differential expression analysis.The function of PdMYB2 was analyzed by subcellular localization,qRT-PCR,transient overexpression,transgenic tobacco and LC-MS/MS.[Result]A total of 64 MYB genes were identified,including 25 R2R3-MYBs,one R1R2R3-MYBs,two 4R-MYBs and 36 members of MYB-related.The PdMYB2 was found to express in the nucleus and cytoplasm.Overexpression of PdMYB2 in tobacco increased anthocyanin accumulation and up-regulated the expression of anthocyanin-related structural genes NtCHS and NtCHI in petals and leaves.[Conclusion]PdMYB2 plays a positive regulatory role in the flavonoid biosynthesis pathway,which can promote the accumulation of anthocyanins and increase the expression of structural genes in the flavonoid pathway.

关 键 词：滇牡丹 花色 MYB转录因子家族 PdMYB2 功能验证 

分 类 号：S685.11[农业科学—观赏园艺]