普通烟草CESA基因家族成员的鉴定、亚细胞定位及表达分析

发布时间：2018-03-23 21:43

本文选题：普通烟草　切入点：CESA基因家族　出处：《遗传》2017年06期

【摘要】：纤维素合成酶蛋白(cellulose-synthase proteins,CESA)是一类质膜定位蛋白,以蛋白复合体的形式存在于质膜上合成纤维素,在细胞壁建成和植物生长发育过程中起着非常重要的作用。本研究利用CESA蛋白保守域序列PF03552检索普通烟草(Nicotiana tabacum L.)蛋白序列,并通过拟南芥(Arabidopsis thaliana)10个CESA蛋白序列在普通烟草基因组数据库中利用TBLASTN程序进行比对,共获得21条NtCESA基因候选序列,对这些序列进行蛋白序列理化性质分析、系统进化树构建、基因结构分析、保守结构域及跨膜区分析和组织表达模式分析,并对NtCESA9和NtCESA14两个蛋白进行了亚细胞定位实验。结果表明:获得的21条NtCESA蛋白序列的理化性质相似;系统进化分析将21个NtCESA基因和10个At CESA基因分成5个分支,每一个分支各成员之间的进化相对保守,基因结构类似,不同分支之间的基因结构差异也较小;NtCESA蛋白结构域相对保守,都含有CESA蛋白典型的N端锌指结构、C端跨膜区和DDD-QXXRW保守功能域;组织表达分析结果表明,大部分NtCESA基因在幼苗和成熟期烟草的根、叶、胚芽和愈伤组织中都有表达,同一个分支中的基因表达模式基本一致,并且NtCESA基因参与初/次生细胞壁纤维素的合成与该基因编码蛋白的跨膜区数目存在关联,表明NtCESA基因家族成员功能上的复杂性;亚细胞定位结果证实NtCESA9和NtCESA14为质膜定位蛋白。本研究为烟草CESA基因家族功能的深入研究奠定了基础。
[Abstract]:Cellulose synthase (Cellulose-synthase proteinsCesa) is a kind of plasmalemma localization protein, which exists in the form of protein complex on the plasma membrane to synthesize cellulose. It plays an important role in cell wall building and plant growth and development. In this study, the conserved domain PF03552 of CESA protein was used to search the protein sequence of Nicotiana tabacum L.in common tobacco. Ten CESA sequences of Arabidopsis thaliana were compared by using TBLASTN program in the general tobacco genome database. A total of 21 candidate sequences of NtCESA gene were obtained. The physicochemical properties of these sequences were analyzed and the phylogenetic tree was constructed. Gene structure analysis, conserved domain and transmembrane domain analysis, tissue expression pattern analysis, and subcellular localization of NtCESA9 and NtCESA14 proteins were carried out. The results showed that the obtained 21 NtCESA protein sequences had similar physicochemical properties. Phylogenetic analysis divided 21 NtCESA genes and 10 at CESA genes into five branches. The evolution of each member of each branch was relatively conserved, and the gene structure was similar. The structural differences of NtCESA protein between different branches are also relatively conserved, both of which contain the typical N-terminal zinc finger structure, C-terminal transmembrane region and DDD-QXXRW conserved functional domain, and the results of tissue expression analysis indicate that, Most of the NtCESA genes were expressed in root, leaf, embryo and callus of seedling and mature tobacco, and the same gene expression pattern was found in the same branch. Moreover, NtCESA gene involved in the synthesis of primary / secondary cell wall cellulose was associated with the number of transmembrane regions of the protein encoded by the gene, indicating the functional complexity of the members of the NtCESA gene family. The results of subcellular localization confirmed that NtCESA9 and NtCESA14 were plasma membrane localization proteins, which laid a foundation for the further study of the function of CESA gene family in tobacco.
【作者单位】：中国农业科学院烟草研究所中国农业科学院烟草遗传改良与生物技术重点开放实验室;中国农业科学院研究生院;
【基金】：国家科技支撑计划项目(编号:2015BAD15B03-05) 国家自然科学基金项目(编号:31470291,31670302)资助~~
【分类号】：Q943.2

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