Identification And Analysis Of Interaction Proteins Of Soybean Stress Resistant Protein PM18

Late embryogenesis abundant(LEA)protein is intrinsically disordered and related to stress resistance in plant.Under stress condition,LEA protein could protect proteins(enzymes),lipids or cell membrane structures,and bind metal ions.According tothe conserved amino acid region,LEA protein can be divided into seven groups.The soybean anti-reverse proteinPM18 belongs to the group 3 LEA family(LEA3).Previous studies have shown that the PM18 protein is resistant in vitro and in Arabidopsis.It also exist alternative splicing in PM18,which has two transcripts(PM18A and PM18B).The interaction proteins of PM18 A and PM18 B are screened by yeast two hybrids.The main content of this paper is to confirm the interaction protein of PM18 A and PM18 B,then analyze the differential expression of these genes in soybean stem and leaf.The specific results are as follows:the full-length rotary validation of yeast,Pull-down,and BiFC were used to verify the interaction between PM18A/B and candidate interaction proteins.It was found that PM18B-9can interact with PM18A/B.PM18B-4 can interact with PM18A/B in vitro,and PM18A-1 has interaction with PM18 A in vitro.the subcellular localization of PM18A/B was then detected.Finally,qRT-PCR was used to study the transcription level of PM18 and the coding genes of interacting protein under salt stress.The relative expressions of PM18B-9,PM18 A and PM18 B genes were up-regulated in salt-stressed soybean leaves and stems,and the level of gene expression were up-regulated more obviously in soybean stems.In addition,the relative expression of PM18B-4 in salt-stressed soybean leaves was down-regulated.The upregulation of expression in salt-stressed soybean stems indicated that PM18B-4 had tissue differences in response to stress.In summary,the interaction protein of PM18A/B was identified,and the molecular mechanism of PM18A/B under abiotic stress in soybean were analyzed in this paper.