河北杨脱水素基因(Phdhn1)的克隆与胁迫表达分析

Cloning and Expression Analysis of Dehydrin Gene(Phdhn1) in Populus hopeiensis Under Stress

为了鉴定河北杨中的脱水素基因并研究其在逆境胁迫中的表达规律,以初步分析脱水素基因在河北杨逆境胁迫中的调控功能,在实验室前期工作的基础上设计引物,通过PCR方法扩增河北杨cDNA文库获取脱水素基因,并通过半定量RT-PCR方法研究其在逆境中的表达情况。结果表明,河北杨脱水素基因全长856 bp,共编码227个氨基酸。同源性分析表明,河北杨中的脱水素基因与已知的其他杨树脱水素高度同源,因此将该基因定名为Phdhn1。半定量RT-PCR结果表明在干旱、低温、高温、高盐及ABA溶液不同时间段(0、0.5、1、2、5、10、24 h)的胁迫下,河北杨Phdhn1在干旱、ABA以及低温胁迫下均出现表达差异。该研究初步阐述了Phdhn1的逆境表达模式,说明了Phdhn1在河北杨逆境胁迫中起到的重要作用。

In order to identify the dehydrin gene of Populus hopeiensis and analyze the expression regularity under abotic stresses for the genetic regulation functions elucidation in stress,the primers were designed on the basis of previous work in our lab and the gene was amplified by PCR using cDNA library,the expression of the gene under abotic stresses was analyzed by semi-quantitative RT-PCR.The results indicated that dehydrin gene of P.hopeiensis was 856 nucleotides long and encoded a protein of 227 amino acid sequences. Sequencing and the homology analysis indicated that P.hopeiensis dehydrin gene was highly similar with dehydrin gene in Populus species.Consequently P.hopeiensis dehydrin gene could be named Phdhn1. Semi-quantitative RT-PCR expression analysis of Phdhn1 under drought,cold,heat,high salinity and ABA at different stress stage（0,0.5,1,2,5,10,24 h） showed that it was differentially expressed in response to drought,ABA and cold stresses.The study showed that Phdhn1 played an important role in abotic stresses of P. hopeiensis.