转BADH基因花生幼苗抗盐性研究

Study on the Salt Tolerance of Transgenic Peanut Seedlings Containing BADH Gene

[目的]为提高花生幼苗的抗盐性和BADH基因在花生耐盐基因工程中的应用提供试验依据。[方法]以根癌农杆菌LBA4404为转化受体菌,pCGⅡ(Kmr)为表达载体,将BADHcDNA导入花生,并通过测定盐胁迫下花生幼苗的根冠鲜重、细胞膜相对电导率和叶绿素含量研究转化植株的耐盐性。[结果]PCR检测结果表明转化植株有1 301 bp目的带,阴性对照植株无目的带。对阳性植株进行Northern杂交,结果表明BADH基因已整合到转化植株的基因组中,并可以正常表达。盐胁迫下,检测组植株的根冠鲜重比对照组植株显著增加;检测组植株的相对电导率与对照组植株差异显著,转基因植株的相对电导率均低于对照植株;对照组植株的叶绿素含量比检测组植株低且差异显著。[结论]转BADH基因花生比对照植株表现出更强的耐盐性。

[ Objective ] The aim was to supply experimental foundation for enhancing the sah tolerance of peanut seedlings and applying BADH gene in the salt-resistant gene project of peanut. [Method] The BADH cDNA was mediated into peanut with agrobacterium tumefaciens LBA4404 as transfomation receptor and pCGⅡ （Kmr） as expression vector and the salt tolerance of transformed plants was studied through determining the fresh weight of root cap, relative conductivity of cell membrane and content of chlorophyll of peanut seedlings under salt stress. [Result] The results of PCR detection showed that the transformed plants had 1 301 bp target band and the negative control plants had no target band. The positive plants were hybridized by Northern hybridization and the results showed that BADH gene had been integrated into the genomes of transfomed plants and it could be expressed normally. Under salt stress, the flesh weights of root cap of plants in detection groups were increased significantly in comparison with the control group, their relative canductivities had significant difference from that in the control group, the restive conductivities of transgenic plants were lower than that of the control plants, the contents of chlorophyll in plants in the control group were lower than that in plants in the detection group and their difference was significant. [Conclusion] Compared with control plants, the peanut containing BADH gene showed stronger salt tolerance.