羊草水孔蛋白基因LcPIP的克隆与表达分析  

Cloning and Expression Analysis of LcPIP cDNA from Leymus chinensis

作　　者：张旸[1] 孙天旭[1] 汤明威 郭勃予 解莉楠[1] 李玉花[1] 

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040

出　　处：《植物生理学报》2014年第10期1585-1592,共8页Plant Physiology Journal

基　　金："十二五"农村领域国家科技计划课题项目(2013AA 102706)

摘　　要：本研究基于羊草水孔蛋白EST序列，应用RACE克隆技术从盐胁迫的羊草中克隆了cDNA全长为1204bp的水孔蛋白基因，其GenBank登录号为KJ459872。经生物信息学分析，该基因开放阅读框为879bp，其编码的蛋白含有292个氨基酸，蛋白质分子量为30．93kDa，理论等电点（pI）为7．00，与已知的小麦、大麦和玉米等单子叶植物来源的同类基因同源性较高，相似性为80％～98％，与小麦TaPIP1的遗传关系最近，与PIP1家族聚为一类。荧光定量PCR分析显示，在200mmol·L^-1NNa，CO3胁迫不同时间后，羊草根LePIP基因的表达量在6h时受到抑制，12～24h之间表达量明显上升，但胁迫持续48h后，LcPIP表达量降至极低水平。羊草叶片的LcPIP基因表达量逐渐上升，48h达到最高峰，72h表达量下降。Based on the PIP EST sequence ofLeymus chinensis, a full length cDNA named LcPIP from L. chin- crisis under salinity stress was cloned by RT-PCR RACE. Sequence analysis indicated that the gene contained 1 024 bp nucleotide, and an open reading frame （ORF） encoding 292 amino acids. The molecular weight of the protein was 30.93 kDa and theoretical pI was 7.00. Homology analysis showed that the deduced amino acid sequence of LcPIP shared 85%-98% identity with PIP from monocotyledon, such as Triticum aestivum, Hor- deum vulgare and Zea mays, had a closest genetic relationship with wheat TaPIP1, belonged to the PIP1 fam- ily. The expression of LcPIP was analyzed by fluorogenic quantitative PCR under Na2CO3 stress. With the extension of Na2CO3 stress time, the expression of LcPIP dropped from 0 to 6 h in the roots, and then de- creased, the peak of expression level arrived at 12 h; but dropped after 24 h. In the leaves, the expression of LcPIP decreased gradually, and the peak of expression level arrived at 48 h; and then the LcPIP expression dropped after 48 h.

关 键 词：RACE 质膜内在蛋白 生物信息学 羊草 表达分析 

分 类 号：S543.9[农业科学—作物学]