番茄质膜H^+-ATPase家族基因的鉴定和表达分析  

Identification and expression analysis of tomato plasma membrane H^+-ATPase family genes

作　　者：刘健健[1] 刘俊丽[1] 季敏杰 陈家栋[1] 杨晓峰[1] 陈爱群[1] 

机构地区：[1]南京农业大学资源与环境科学学院,江苏南京210095

出　　处：《浙江农林大学学报》2016年第5期734-741,共8页Journal of Zhejiang A&F University

基　　金：国家自然科学基金资助项目(31372121)

摘　　要：植物质膜H^+-ATPase(Ec3.6.1.3)是一类普遍存在于细胞质膜上通过水解三磷酸腺苷(ATP)产生能量,将细胞质中的氢离子(H^+)逆浓度泵出细胞的运输蛋白。植物中的质膜H^+-ATPase由一个多基因家族所编码,其功能涉及到植物生长发育的多个生理过程。通过对全基因组检索在茄科Solanaceae植物番茄Solanum lycopersicum中共鉴定到8个编码质膜H^+-ATPase的同源基因(LHA1~8)。生物信息学分析显示:这8个LHA基因具有较高的序列相似性和较为保守的外显子/内含子结构特征。实时荧光定量聚合酶链式反应(q RT-PCR)分析显示,LHA1~4在所有被检测的组织器官中都有表达,LHA5~7几乎只在花器官中高量表达,而LHA8在正常培养和养分(氮、磷、钾和镁)缺乏以及高盐胁迫处理条件下几乎都不表达,但能够在被菌根真菌侵染的根系中强烈表达。将一段2 669 bp的LHA8的启动子融合GUS报告基因转入到烟草Nicotiana tabacum中发现,GUS基因几乎只在被菌根真菌菌丝侵入形成丛枝的根系细胞中特异性表达。Plasma membrane(PM) H^+-ATPase, which catalyzes ATP hydrolysis coupled with pumping out protons from cells and which is ubiquitously existent in all cell types of plants examined so far, has been characterized to be encoded by a multigene family involved in a number of plant physiological and developmental processes. The aim of this study is to identify and characterize the genes encoding PM H^+-ATPase in Solanum lycopersicum(tomato). Through genomic sequence database hunting and bioinformatics analysis, eight putative PM H^+-ATPase genes(named as LHA1-8) were identified, and their expression patterns were subsequently analysed by quantitative real-time reverse transcription polymerase chain reaction(q RT-PCR). Bioinformatics analysis revealed a high conservation among these paralogous genes in both coding sequences and Exon/Intron structures. Expression analysis showed that LHA1, LHA2, LHA3, and LHA4 were expressed in all tissues examined; whereas LHA5, LHA6, and LHA7 were predominantly expressed in flowers. LHA8, the newly identified gene in this study, with barely detectable transcripts in normal growth conditions or in nutrient(N, P, K,and Mg)-deficient conditions, was strongly induced in arbuscular mycorrhizal(AM) fungal-colonized roots. A2 669 bp LHA8 promoter fragment could direct the β-glucuronidase(GUS) reporter expression specifically in transgenic tobacco roots colonized by AM fungi. The results provide new insights into the evolutionary conservation and functional divergence of the tomato PM H^+-ATPase gene family during plant growth and development.

关 键 词：分子生物学 番茄 质膜H+-ATPase 表达模式 菌根真菌 

分 类 号：S641.2[农业科学—蔬菜学] Q943.2[农业科学—园艺学]