月季Rchsp1718基因转化烟草的非生物胁迫耐性研究

 为了研究从月季中分离的小分子热激蛋白Rchsp1718的生理功能, 把Rchsp1718的编码框插入到表达载体PHB的35S组成型启动子后面, 通过农杆菌GV3101转化烟草, 获得转基因植株。在高温、干旱、渗透、高盐胁迫条件下分别检测转基因烟草形态、失水率、电导率、脯氨酸含量及其内外源基因表达模式。结果显示: 与对照相比, 转基因烟草抗性表型明显, 电导率较低, 脯氨酸含量大幅度提高, 失水率降低; P5CS基因的表达量上调, 说明转Rchsp1718基因烟草苗在高温以及高渗透胁迫下表现出明显的耐受性。

Transformation of Tobacco with Rchsp1718 from Chinese Rose EnhancesTolerance to Different Abiotic Stresses

Rchsp1718 cloned from Chinese rose belongs to a cytosolic class I sHSP, and it is induced under different stress. In order to study the physiological function of Rchsp1718, we subcloned the ORF of Rchsp1718 recombined after 35S: p romoter into DH5α, then transformed into plant gene expression vector Agrobaterium rumefaciens, leaf disc of tobacco was infected by Agrobaterium and three transformants were attained. We assessed the phenotype, relative electrical leakage, proline content, water lose rate under high temperature, osmotic stress, and salt stress respectively, and the transcrip t levels of P5CS gene was also investigated by semi-quantitative RT2PCR. Results showed that transgenic line showed lower relative electrical leakage, more proline content, lower water lose rate, and the expressed of P5CS was elevated. Comparedwith the non-transgenic lines, transformed ones showed obvious resistance to high temperature and osmotic stress. The possible mechanism to different abiotic stresswas also discussed.