论文部分内容阅读
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-1的Na2CO3胁迫不同时间后,羊草根中LcPIP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。
In this study, a 1204bp aquaporin gene was cloned from Leymus chinensis using RACE cloning technology. Its GenBank accession number is KJ459872. Bioinformatics analysis revealed that the open reading frame (ORF) of this gene was 879bp. The encoded protein contained 292 amino acids with a molecular weight of 30.93kDa and a theoretical isoelectric point (pI) of 7.00. Compared with the known list of wheat, barley and maize The homologous genes of the same plant originated from leaves are higher, the similarities are 80% ~ 98%. The genetic relationship with TaPIP1 is the most recent, which is similar to PIP1 family. Fluorescence quantitative PCR analysis showed that the expression of LcPIP gene was inhibited at 6h after 200mmol·L-1 Na2CO3 stress, and the expression level of LcPIP increased significantly after 12 ~ 24h, but the LcPIP The amount of expression dropped to very low levels. The expression of LcPIP gene in Leymus chinensis leaves increased gradually, reached its peak at 48h and decreased at 72h.