水稻OsNAD-ME1的克隆、预测及逆境下的表达分析  

Cloning and Prediction of OsNAD-ME1 from Oryza sativa L. and Its Expression Profiles under Stresses

作　　者：周滈[1] 杨传平[1] 柳参奎[1] 

机构地区：[1]林木遗传育种与生物技术国家重点实验室,盐碱地生物资源环境研究中心,东北林业大学,哈尔滨150040

出　　处：《分子植物育种》2011年第6期680-687,共8页Molecular Plant Breeding

摘　　要：植物中的苹果酸酶能够参与光合作用、呼吸作用和脂类生成等多种重要的代谢途径,而且与植物的逆境防御有着密切的关系。为研究水稻中一个苹果酸酶基因(OsNAD-ME1,GeneID:4343294)的基本特征及其与逆境间的响应关系,克隆得到该基因的cDNA片段,应用生物信息学对其序列进行分析及预测,并运用Northern检测的方法,对该基因在多种非生物逆境下的表达情况做出研究。由序列分析结果可知,OsNAD-ME1的ORF为1869bp,编码622个氨基酸,OsNAD-ME1属于植物NAD-ME的α亚组,其理论分子量为68.9kD,理论等电点为6.03。预测结果表明该蛋白属于苹果酸酶家族,无跨膜结构域,定位于线粒体上。并利用GFP定位的方法,在拟南芥悬浮细胞中对亚细胞定位的预测结果进行了验证。Northern检测结果显示,OsNAD-ME1能够响应多种非生物逆境,在NaCl和NaHCO3胁迫下其表达水平在24h时达到最大,在PEG和H2O2胁迫下其表达水平在12h时达到最大。该结果表明OsNAD-ME1可能与植物对非生物胁迫的响应有关。Malic enzymes in plants are involved in diverse important metabolic pathways, such as photosynthesis, respiration and lipid biosynthesis. And it was found that malic enzymes were involved in plant defense responses. In order to investigate the characters of a rice malic enzyme gene （OsNAD-ME1, Gene ID： 4343294） and confirm the response relationship between this gene and stress environments, OsNAD-ME1 was cloned and bioinformatics analysis and northern analysis of the response to stress environments about this gene were performed. The results of sequence analysis indicated that the open reading frame of OsNAD-ME1 was 1 869 bp in length, encoding 622 amino acid residues. OsNAD-ME1 belonged to α group of plant NAD-ME with predicted molecular weight of 68.9 kD and isoelectric point of 6.03. OsNAD-ME1 was a member of malic enzyme family, whereas it didn＇t have transmembrane domain. Sub-cellular location prediction showed that OsNAD-ME1 was located in mitochondria, and the predicting result was verified in Arabidopsis protoplasts using green fluorescence protein to track the location of this protein. The results of northern analysis indicated that OsNAD-ME1 had response to several abiotic stresses. In the presence of NaCl and NaHCO3, transcripts of OsNAD-ME1 reached peak level at 24 h of treatment, and the expression of OsNAD-ME1 was induced drastically at 12 h under PEG and H2O2 stresses. The results indicated that OsNAD-ME1 was involved in response to abiotic stresses.

关 键 词：OsNAD-ME1 序列分析 亚细胞定位 Northern检测 

分 类 号：S511[农业科学—作物学]