Study On The Difference Of Secreted Proteins In Chestnut Blight Fungus And The Functions Of Protein

The study of secretion mechanism and pathways in filamentous fungi Cryphonectria parasitica remain at early stage.Here,adopting virulent wild-type strain EP155,hypo virus CHV1-EP713 infected hypovirulent strain EP713,hypovirus CHV1-Euro7 infected moderate hypovirulent strain Euro7 and the papain-like protease infectious cDNA clone p29 infected strain P29 as experimental material,we used iTRAQ technology to quantify the differential secretory proteins between EP155 and the other three hypovirulent-related strains,in order to understand how the hypovirus affects the secreted proteins and virulence factors.In three independent expriments,755 proteins were identified in which 43 proteins were up-regulated and 48 proteins were down-regulated in EP713 comparing to EP155;747 proteins were identified in which 16 proteins were up-regulated and 14 proteins were down-regulated in Euro7 comparing to EP155;629 proteins were identified in which 4 proteins were up-regulated and 19 proteins were down-regulated in P29 comparing to EP155.For further understanding the roles of differential secretory proteins,we constructed the null mutations of five proteins including laccase(id:327261),FAD binding domain containing protein(id:350207),aspartyl protease(id:107093),and two hypothetical proteins(id:351611,356341).The deletion of these five genes resulted in abnormal morphology,reduced pigmentation,less aerial hyphae and sporulation.Furthermore,the deletion of 107093 lead to reduced virulence.These results indicate that the five secretory proteins are related to growth,sporulation,pigmentation and virulence in Cryphonectria parasitica.Our study provided a solid foundation for study of secretory pathway and regulation mechanism in filamentous fungi..