Functional Study Of AfphyA(b) Gene In Soybean (Glycine Max Merr.) And Bioinformatics Analysis Of The Thpht1 Gene Family

Phosphorus is an important nutrient element involved in the physiology and growth of soybean.But the mobility of phosphorus is poor in the soil,it is difficult for phosphorus to be absorbed for its adsorption.So it turns to be the focus of research on genetically modified soybean to enhance the ability of crops to transport the phosphorus and improve the efficiency of phosphorus absorption.According to the preference of codon usage,the gene phyA?b?of Aspergillus fumigatus which is suitable for expression in soybean was synthesized by codon optimization of phytase gene phyA.The plant expression vector pCBPS-phyA?b?was constructed with pCAMBIA3301 as the backbone,which is regulated by the agglutinin gene promoter and signal peptide sequence.Agrobacterium tumefaciens-mediated transformation of Jilin 35 soybean cotyledon,the results showed that the target gene had been integrated into the soybean genome by PCR detection,bar test strip test and herbicide leaf smear detection.The highest amount of exogenous gene T₃ strain expression was selected according to the results of the RT-PCR,which were used in the progress of correlation analysis of phosphorus.The results showed that phyA?b?overexpression lines was superior to neigher phyA overexpression lines nor wild type in many aspects such as phytase activity,phytate phosphorus content and inorganic phosphorus content.It was concluded that the phytase expressed through the synthetic phyA?b?gene was more active than the unmodified one in soybean seeds,which indicated that codon optimization could improve the expression of exogenous genes.Phosphate transporters is a kind of protein which can transfer the inorganic phosphorus into available phosphorus that can be absorbed by the plant easily.The process of phosphorus uptake by plants is the same as the process of the transportation of the plasma membrane mediated by the phosphate transporter.Therefore,it is of great theoretical and practical significance to identify and isolate the genes related to the progress of uptake and utilization of phosphate.According to the phosphate transporter gene with known function in Arabidopsis thaliana,the phosphate transporter genes were found through Phytozome website,the30 gene sequences that has the highest functional similarity were chosen as a result of comparison.It was found that the gene structure of the PHT family was more conserved than that of the PHO gene family by analyzing the intron and extron structure of phosphate transporters.The results of the conserved motifs of the Thellungiella halophila phosphate transporter showed that the stability and conservativeness of subfamily ThPHT1 genes are higher than those of other subfamilies of PHT.Therefore,this paper focuses on systematic analysis of PHT1subfamily members by analyzing the results of multiple sequence alignment and the spectrum of phylogenetic tree of ThPHT1 and AtPHT1 subfamily members,the physical and chemical parameters,the hydrophobic of ThPHT1 genes,the secondary and the three-dimensional structures and the subcellular localizations of ThPHT1subfamily members,which further clarified the phosphate transporter gene function and laid the foundation for screening of the functional gene could have efficient use of phosphorus.