Excavation Of Differentially Expressed Genes And Cloning And Transformation Of GmPHF1 Gene Under Low Phosphorus Stress In Soybean

Soybean is a phosphorus-loving crop,and phosphorus deficiency is one of the main factors limiting soybean yield.Although phosphorus is abundant in soil,the amount of phosphorus available for crop uptake in soil is very limited.Although phosphorus fertilizer can alleviate soil phosphorus deficiency,phosphate rock is scarce,and a large amount of fertilization pollutes the environment.Therefore,in order to clarify the ways of phosphorus uptake and transport by high phosphorus efficient genotypes,the following experiments were carried out to improve the phosphorus utilization efficiency of soybean in soil.Soybean varieties with high phosphorus efficiency and low phosphorus efficiency were treated with normal phosphorus(P=0.5mmol/L)and low phosphorus(P=0.005mmol/L)respectively.The results showed that there were significant differences in root morphology,phosphorus content and free amino acid content between the two types of soybean varieties.Under low phosphorus treatment,the total root length of P-efficient soybean varieties was longer,the number of root tips was more,the free amino acid content of P-inefficient soybean varieties increased,and the phosphorus content of roots of both types of soybean varieties decreased.A total of 9669 differentially expressed genes were obtained from the transcriptome sequencing data of the roots of two soybean cultivars under low phosphorus stress.236 genes were found to be involved in the response to low phosphorus stress,of which 10 genes were related to low phosphorus stress.The PHF1 gene was further studied.PHF1 is a phosphate transporter absorption and transport related gene.The CDs of GmPHF1 are 1185 bp,encoding 394 amino acids.The expression patterns of GmPHF1 in soybean tissues showed that GmPHF1 gene was mainly expressed in soybean roots and leaves.The plant overexpression vectors pCAMBIA3300N-GmPHF1 and pCNG-GmPHF1-GFP were cloned and constructed.Subcellular localization results showed that GmPHF1 gene was located in the endoplasmic reticulum.Transgenic plants of T2 generation of Arabidopsis thaliana overexpressing GmPHF1 gene were preliminarily obtained.