生姜NHX基因家族成员鉴定及其在硅缓解盐胁迫中的表达特征  

Identification of Ginger(Zingiber officinale Roscoe)NHX Gene Family Members and Characterization of Their Expression Patterns in Silicon Alleviating Salt Stress

作　　者：尹军良 李婧怡 韩硕 杨培华 马佳伟 刘奕清 胡海骏 朱永兴 YIN JunLiang;LI JingYi;HAN Shuo;YANG PeiHua;MA JiaWei;LIU YiQing;HU HaiJun;ZHU YongXing(College of Agriculture,Yangtze University,Jingzhou 434025,Hubei;College of Horticulture and Gardening,Yangtze University/Hubei Key Laboratory of Spices&Horticultural Plant Germplasm Innovation&Utilization/Engineering Research Center of Wetland Ecology and Agricultural Use,Ministry of Education,Jingzhou 434025,Hubei)

机构地区：[1]长江大学农学院,湖北荆州434025 [2]长江大学园艺园林学院/香辛园艺植物种质创新与利用湖北省重点实验室/湿地生态与农业利用教育部工程研究中心,湖北荆州434025

出　　处：《中国农业科学》2024年第19期3848-3869,共22页Scientia Agricultura Sinica

基　　金：湖北省重点研发项目(2022BBA0061);重庆英才优秀科学家项目(CQYC20220101514);湿地生态与农业利用教育部工程研究中心开放基金(KFT202305);重庆市自然科学基金项目-面上项目(CSTB2022NSCQ-MSX1588)。

摘　　要：【目的】系统分析生姜NHX基因家族成员的基本特征,探究其在生姜不同组织、处理下的表达模式,为进一步研究ZoNHXs功能提供理论基础。【方法】以拟南芥NHX蛋白序列为参考序列,与生姜基因组进行BLAST比对,筛选出包含Na^(+)/H^(+)exchange保守结构域的蛋白序列,得到生姜ZoNHX家族成员。使用MEGA 7.0构建拟南芥和生姜NHX家族成员的系统发育树,利用ExPASy ProtParam、GSDS、MEME、PHYRE2、SOPMA和TBtools对ZoNHXs的理化性质、基因结构、保守基序、二级结构和三级结构、染色体定位、基因共线性及表达模式进行分析。借助RT-qPCR分析ZoNHXs在对照(CK)、盐胁迫(NaCl)、盐胁迫+纳米硅(NaCl+SiNP)处理的生姜根系和叶片中的表达特征,并测定生姜各个组织内的Na^(+)、K^(+)离子含量。【结果】从生姜基因组中共鉴定出15个ZoNHX基因家族成员,根据其染色体位置分别命名为ZoNHX1—ZoNHX15。根据系统发育关系和亚细胞定位分析,将ZoNHXs归为液泡膜(vacuole,Vac)、核内体膜(endosome,Endo)和质膜(plasma membrane,PM)3个亚组。蛋白质特征分析结果表明,ZoNHXs的相对分子量介于26.01—163.59 kDa,氨基酸序列长度介于231—1 459aa。亚细胞定位分析发现,11个ZoNHXs定位在液泡,ZoNHX14定位于细胞膜和细胞核,ZoNHX1、ZoNHX7和ZoNHX9分别定位于细胞质、细胞膜和叶绿体。信号肽预测发现,只有ZoNHX1为含有信号肽的分泌蛋白,其余均为非分泌蛋白。顺式作用元件分析显示,ZoNHXs启动子区域包含生长发育响应元件、激素响应元件和胁迫响应元件。转录组数据分析表明,15个ZoNHXs在生姜叶片和根系中均有表达;其中,ZoNHX12在生姜各组织和各非生物胁迫下表达水平相对较高,ZoNHX14在生姜各生长时期及低温下表达量显著上调。RT-qPCR结果显示,与CK相比,除ZoNHX8、ZoNHX11外,其余基因在盐胁迫后的根系和叶片中表达量均显著上调。同单独盐胁迫相比,NaCl+Si【Objective】Systematic analysis of the basic features of the ZoNHX family members in ginger and exploration of their expression patterns in different tissues and treatments of ginger can provide a theoretical foundation for further research on the function of ZoNHXs.【Method】The Arabidopsis thaliana NHX protein sequences were used as reference sequences to compare with the ginger genome to obtain the ginger ZoNHXs.The phylogenetic tree containing Arabidopsis and ginger NHX family members were constructed based on the neighbor-joining method by MEGA 7.0.Physicochemical properties,gene structure,conserved motifs,secondary structure and tertiary structure of ZoNHXs were analyzed by using ExPASy ProtParam software,GSDS,MEME online website,PHYRE 2 and SOPMA,while chromosomal localization,intragenomic covariance and expression patterns of ZoNHXs were analyzed by using TBtools software.The gene expression levels of ZoNHXs in different treatments including CK,NaCl,and NaCl+SiNP100,and different ginger tissues were analyzed using RT-qPCR.The Na^(+)and K^(+)contents of ginger tissues was determined.【Result】A total of 15 NHX gene family members were identified from the ginger genome,and were named as ZoNHX1-ZoNHX15 based on their chromosomal locations.The ZoNHXs were classified into three subgroups according to phylogenetic relationship and subcellular localization,including vesicular membranes(Vac,vacuole),endosomal membranes in the nucleus(Endo,endosome)and plasma membranes(PM,plasma).Protein characterization analysis showed that the relative molecular weights of ZoNHXs ranged from 26.01 kDa to 163.59 kDa,and the protein lengths ranged from 231 to 1459 aa.The results of subcellular localization prediction showed that 11 ZoNHXs were distributed on the vacuole,ZoNHX14 were distributed both on the cell membrane and nucleus,ZoNHX1,ZoNHX7 and ZoNHX9 were respectively distributed on cytoplasm,cell membrane,and chloroplast.Signal peptide prediction showed that only ZoNHX1 contained a signal peptide and was a secreted p

关 键 词：生姜 NHX基因家族 盐胁迫 表达分析 转录组 

分 类 号：S632.5[农业科学—蔬菜学]