The Interaction Between ZmHAK1 And ZmCIPK23 Proteins In Maize

The ZmHAK1 protein in maize is a kind of high-affinity potassium transporter proteins.Under a-low amount of potassium stress,it can enhance the ability of maize absorbing and utilizing K~+.In order to explore the molecular mechanism of ZmHAK1 gene and its expressed protein in this function,this paper studied the interaction between ZmHAK1 and ZmCIPK23 proteins.The molecular mechanism of ZmHAK1 gene in promoting the ability of maize absorbing potassium ions under a-low amount of potassium stress was gradually revealed.The mechanism lays a solid foundation for cultivating maize varieties resisting a-low amount of potassium stress,reducing potassium fertilizer application,reducing production costs,and protecting the ecological environment.In this study,the vector of subcellular localization containing ZmCIPK23 gene was constructed by Gateway ligation technology.The vector of fusion protein was transferred into tobacco.The transformed tobacco leaves were observed by fluorescence microscopy and laser confocal microscopy.The results showed that the fluorescent signal of ZmCIPK23 was located on the plasma membrane.In this study,the vector of yeast two-hybrid and the bimolecular fluorescent complementary containing ZmHAK1 and ZmCIPK23 genes were constructed by Gateway technology,and the interaction between ZmHAK1 and ZmCIPK23proteins was studied by yeast two-hybrid system and bimolecular fluorescence complementation technology.In the yeast two-hybrid experiment,no colonies were observed on the quadruple dropout.The results showed that there was no interaction between the two proteins.In the bimolecular fluorescence complementation experiment,the infected tobacco leaves were observed by fluorescence microscope.There was no fluorescence signal of the blank control which was transferred to the blank vector;on the contrary,there was a strong fluorescence signal on the plasma membrane of the experimental group which was co-transferred into the expression vector containing the target genes.The results indicated that there was an interaction between ZmHAK1 and ZmCIPK23 proteins.The RNA interference vector pRNAi-ZmCIPK23 of ZmCIPK23 gene was transfected into maize inbred lines,and the positive maize seedlings with RNA interference vector pRNAi-ZmCIPK23 were screened by Basta spraying and genomic DNA PCR.