拟南芥HKT1基因pCAMBIA载体构建及转基因材料筛选

Construction of pCAMBIA vector of Arabidopsis thaliana HKT1 gene and identification of transgenic lines

探索盐胁迫作用机理并将其运用到农业生产领域中,对培育耐盐作物具有非常重要的现实意义。土壤环境中过量的Na+会干扰植物根对其他离子的吸收,导致植物生理代谢紊乱、离子失衡。在拟南芥中,已有研究证明HKT1基因在盐胁迫信号通路中扮演十分重要的角色,控制着Na+的吸收、转运和储存。文章通过正向和反向遗传学手段,发现一个转录因子MYBX在分子水平正向调控基因HKT1的表达,并通过构建HKT1过表达载体、获得含有测序正确质粒的农杆菌、侵染拟南芥突变体mybx,最终筛选获得1300-HKT1/mybx转基因植株,从而可以进一步验证MYBX和HKT1之间的关系。

Exploring the signaling pathway of salt stress and applying it to the field of agricultural production have very important practical significance for the cultivation of salt-tolerant crops.Excessive Na+in the soil environment can interfere with the absorption of other ions by plant roots,leading to plant physiological and metabolic disorders and ion imbalances.Studies have shown that HKT1 gene plays a very important role in the salt stress signaling pathway in Arabidopsis thaliana,which controls the absorption,transport and storage of Na+.In the laboratory,through forward and reverse genetics,this paper proposed a transcription factor MYBX that may positively regulate the expression of the gene HKT1 at the molecular level.By constructing HKT1 overexpression vector,obtaining Agrobacterium containing the correct sequencing plasmid,and infecting Arabidopsis mutant mybx,1300-HKT1/mybx transgenic plants were finally screened to further verify the relationship between MYBX and HKT1.