海滨雀稗PvPRMT家族成员鉴定、表达模式及PvPRMT10基因的功能  

Paspalum vaginatum PvPRMT family member identification,expression patterns and PvPRMT10 gene function analysis

作　　者：孙灿灿 刘宇 邹月 刘君 杨志民 陈煜 SUN Cancan;LIU Yu;ZOU Yue;LIU Jun;YANG Zhimin;CHEN Yu(College of Agro-Grassland Science,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学草业学院,江苏南京210095

出　　处：《南京农业大学学报》2021年第4期705-715,共11页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(31672193)。

摘　　要：[目的]蛋白质精氨酸甲基转移酶(protein arginine methyltransferases,PRMT)家族基因在调控植物生长发育及抗逆途径过程中发挥重要作用,而草坪草PRMT成员的表达模式和抗逆功能尚不明晰,本文旨在分析海滨雀稗PvPRMT基因家族在非生物胁迫下的表达模式并鉴定PvPRMT10成员的抗逆功能。[方法]以暖季型草坪草海滨雀稗为研究材料,依据转录组数据库信息,经序列分析及PCR克隆获得8个海滨雀稗PRMT家族成员全长序列,进行系统进化树及MEME蛋白结构域分析;采用荧光定量PCR分析不同胁迫条件(盐、干旱、重金属镉和ABA处理)下PvPRMT的相对表达水平;通过转基因技术获得PvPRMT10拟南芥过量表达株系,并进行胁迫条件下表型鉴定。[结果]同为单子叶的海滨雀稗与水稻的8个PRMT家族成员进化关系相较于拟南芥更为接近,可以分为3类(Ⅰ型、Ⅱ型和Ⅲ型)。进化关系越近的基因在不同胁迫下的表达模式也越为相似;除PvPRMT4外,其他7个成员基因的表达对不同胁迫均有不同程度的上调或下调,叶中PvPRMT10在NaCl处理下的表达量在6和12 h时增加近10倍,在CdCl_(2)处理下的表达量在6 h时增加近15倍;海滨雀稗叶中PRMTs基因响应比根系更为强烈。进一步转基因发现,Ⅰ型PvPRMT10过表达拟南芥株系在重金属镉(Cd)和盐胁迫下的种子萌发和幼苗生长均优于野生型,CdCl_(2)处理下过表达株系的种子存活率提高15%,NaCl处理下过表达株系的种子存活率提高45%,且幼苗根长提高近2倍。[结论]本研究克隆获取海滨雀稗PRMT家族基因,分析其在非生物胁迫下的表达模式,且发现PvPRMT10具有正调控耐镉和耐盐的功能。[Objectives]Protein arginine methyltransferases(PRMT)family genes performed important roles in regulating plant growth,development and stress tolerance.However,the expression patterns and stress tolerance functions of PRMT members in turfgrass were still unclear.The purpose of this study was to explore the expression patterns of Paspalum vaginatum PvPRMT gene family under abiotic stress and the stress tolerance functions of PvPRMT10.[Methods]With warm-season turfgrass seashore paspalum as the plant material,and by referring to the transcriptome database information,sequence analysis and PCR were used to clone the full-length sequences of 8 members of seashore paspalum PRMT family,and then the proteins were analyzed by phylogenetic trees and MEME online tool.Quantitative real-time PCR(RT-qPCR)was used to analyze the relative expression levels of PvPRMTs under different stress conditions(NaCl,drought,Cd and ABA treatment).Arabidopsis PvPRMT10 overexpressed lines were obtained by transgenic technology,and phenotypic identification was performed under stress conditions.[Results]The analysis found that seashore paspalum and rice were both monocotyledon and their 8 genes of PRMT family evolutionary relationship were closer than Arabidopsis,and could be divided into three categories(Ⅰ,ⅡandⅢtype).The more closely related genes had similar expression patterns under different stresses.In addition to PvPRMT4,other genes were up or down-regulated to different expression levels in response to different stresses.The expression level of PvPRMT10 in leaves under NaCl treatment increased by nearly 10 times at 6 and 12 h,and the expression level under CdCl_(2) treatment increased by nearly 15 times at 6 h.PRMT genes responses in leaves were stronger than those in root under different stress conditions.Further,we also found that overexpression of the typeⅠgene PvPRMT10 enhanced cadmium(Cd)and salt stress tolerance of Arabidopsis.The seed survival rate of overexpressed lines under CdCl_(2) treatment was about 15%higher than

关 键 词：蛋白质精氨酸甲基转移酶 海滨雀稗 转录组数据库 非生物胁迫 

分 类 号：S688.4[农业科学—观赏园艺]