碱茅6-磷酸海藻糖合成酶基因的克隆及其耐逆性分析  

Cloning of a TPS gene and analysis of its function in stress tolerance in Puccinellia tenuiflora

作　　者：李莹[1,2] 柳参奎[1] 

机构地区：[1]东北林业大学盐碱地生物资源环境研究中心,黑龙江哈尔滨150040 [2]黑龙江八一农垦大学寒地作物种质改良与栽培重点实验室,黑龙江大庆163319

出　　处：《草业学报》2015年第1期99-106,共8页Acta Prataculturae Sinica

基　　金：黑龙江省寒地作物种质改良与栽培重点实验开放课题资助项目(CGIC201204);教育部创新团队发展计划(IRT13053);黑龙江省自然科学基金项目(C201406);中央高校基本科研业务费专项资金(2572014BA20);863项目(2013AA102701-7);哈尔滨市自然科学基金(2008RFQXN007)资助

摘　　要：从碱茅酵母cDNA文库中经过NaCl、NaHCO。筛选，均得到长为1153bp碱茅6一磷酸海藻糖合成酶基因（PutTPS）片段。通过3’End cDNA amplification方法获得基因缺失的3’序列，该基因全长3358 bp，编码882个氨基酸。氨基酸序列比较结果表明，PutTPS氨基酸序列与水稻、拟南芥、玉米等多种高等植物的TPS蛋白的同源性高达60％～90％。利用Northern blot技术，研究该基因的组织表达模式及NaCl、NaHCO3胁迫处理下基因的表达模式变化；同时对转pYES2-PutTPS基因酵母菌株进行盐、碱、氧化胁迫、渗透胁迫处理，观察其在逆境下的生存表现。结果表明，PutTPS具有组织表达特异性，其中在根和花中表达量最大；NaCl、NaHCO3会诱导PutTPS基因在根和叶中的上调表达；同时重组酵母菌株对盐碱、氧化、渗透胁迫及干旱等逆境的适应能力显著增强。以上研究结果表明，碱茅PutTPS基因与逆境之间具有一定的应答关系，并在植物适应环境逆境过程中起着重要的作用。Abstract： In this study, a 6-trehalose phosphate synthase gene （TPS） fragment, 1153 bp in length, was isola- ted through NaC1 and NaHCO3 screening from Puccinellia tenuiJTora yeast cDNA library. A 3＇-end cDNA am- plification technique was used to clone PutTPS. The full-length cDNA is 3358 bp containing an open reading frame （ORF） of 882 amino acids. Multiple alignment analysis based on amino acids of TPS proteins of rice, Arabidopsis, maize, and other higher plants revealed that the amino acid homologies were about 60% to 90%. The tissue expression patterns of P. tenuiflora PutTPS and the expression under NaC1 and NaHCO3 stress were further investigated by northern blot. Also the recombinant yeast INVScI （pYES2-PutTPS） and control INVScI （pYES2） were subjected to salt, drought, osmotic and oxidative stresses. PutTPS was highly ex- pressed in roots and flowers; in roots and leaves, NaC1 and NaHCO3 induced up-regulated expression of Put-TPS genes and the recombinant yeast cells showed higher stress resistance than control cells. It was also found that the TPS from alkali grass contributes to salt, drought, osmotic and oxidative resistance. The present study indicated that there was a relationship between PutTPS expression and multiple stresses, and PutTPS may play an important role for P. tenuiflora during adaption to environmental abiotic stress.

关 键 词：碱茅 6-磷酸海藻糖合成酶 基因表达 酵母 

分 类 号：Q943.2[生物学—植物学]