Phosphoproteomics Analysis Of Different Stages Of Sexual Reproduction In Phytophthora Infestans

Phytophthora infestans,belonging to the class Oomycetes,are semi-biotrophic pathogens that cause late blight disease in potatoes and many other related plants.For potatoes,the disease causes losses and management costs more than US $6 billion a year and is a major threat to global food security.Sexual reproduction is an important event in the life cycle of Phytophthora,but the research on the mechanism of sexual reproduction is still insufficient.In this study,P.infestans strains P7723 and HQK8-3 were used as materials.The effects of rye medium(RA),carrot medium(CA),rye medium covered with cellophane(CP),and rye medium covered with polycarbonate membrane on growth and reproduction of P.infestans were tested,The results showed that rye medium covered with WATMAN polycarbonate membrane was the most suitable medium for collecting sexual mycelium.Phosphoproteomics analysis was carried out on different stages of sexual reproduction of P.infestans,strains P7723 and HQK8-3 were cultured in confrontation,mycelia were collected at three time points during the stage of sexual reproduction,which were 0 days(PP0?P group,PP0?H group),3 days(PP3 group)and 9 days(PP9 group),respectively;Label Free technique was used to study the expression profiles of different phosphorylated proteins at different time points in sexual reproduction of P.infestans,and bioinformatics analysis was carried out on the differential protein expression profiles to screen out a group of proteins that might be closely related to sexual reproduction in P.infestans.The results of phosphoproteomics analysis showed that a total of 3938 phosphorylated peptides and2028 proteins were detected.There were 444 different proteins in PP3 group compared with PP0?P group.After GO functional annotation and KEGG pathway analysis at the level 2,6 phosphorylated proteins showing significant differences were screened out,including PITG?00334,PITG?11466,PITG?13426,PITG?13666,PITG?19590,and PITG?21117.One phosphorylated protein PITG?10663 showing significant differences was detected in PP3 group compared with PP0?H group.Six phosphorylated proteins with significant differences were screened in PP9 group compared with PP3 group,including PITG?00397,PITG?06171,PITG?18434,PITG?01036,PITG?03936 and PITG?16833.These proteins are very likely to be involved in the sexual reproduction process of P.infestans and their functions need to be further verified in future work.In this study,the proteins related to the sexual reproduction process of P.infestans were screened by phosphorylproteomics analysis,which promoted the understanding of the signaling pathway in the sexual reproduction of P.infestans,and was of great significance for the more effective prevention and treatment of Phytophthora disease in the future.