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草业学报  2015 

碱茅6-磷酸海藻糖合成酶基因的克隆及其耐逆性分析

DOI: 10.11686/cyxb20150113, PP. 99-106

Keywords: 碱茅,6-磷酸海藻糖合成酶,基因表达,酵母

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Abstract:

从碱茅酵母cDNA文库中经过NaCl、NaHCO3筛选,均得到长为1153bp碱茅6-磷酸海藻糖合成酶基因(PutTPS)片段。通过3'EndcDNAamplification方法获得基因缺失的3'序列,该基因全长3358bp,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Northernblot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。

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