Functional Properties Of UeSho1 And UeMsb2 In Ustilago Esculenta

Ustilago esculenta is a parasitic fungus of Zizania latifoli.The ability of the fungus to infect the plant caused the difference of the phenotype of Zizania latifolia.In the field,Zizania latifolia has three phenotypes: normal Jiaobai,male Jiaobai and gray Jiaobai.Only normal Jiaobai has high nutritional and economic values,while male and gray Jiaobai can affect the cultivation of normal Jiaobai,no economic value,so it needs to be cleaned up in time.When both Zizania latifolia and Ustilago esculenta are in the suitable growth environment,it is beneficial to the growth of Zizania latifolia,and the adaptation to the external environment will be through the MAPK signaling pathways.Sho1 and MSb2 proteins were found in Saccharomyces cerevisiae,which were located in the front of FG and HOG-MAPK signaling pathways and involved in filamentous growth and osmotic regulation.In this study,the UeSho1 and UeMsb2 genes were cloned.The biological functions of UeSho1 and UeMsb2 genes were studied.The environmental response of UeSho1 and UeMsb2 genes was evaluated assessment of stress tolerance.Further,in order to investigate the effects of UeSho1 and UeMsb2 deletion on proliferation,filamentation and pathogenic-related genes expression of Ustilago esculenta by q RT-PCR.The UeSho1 was cloned.Sequence analysis results showed that the gene had a total length of 1137 bp,with 1 intron and 2 exons forming 795 bp open reading frame,encoding 264 amino acids,containing SH3 conserved domain.The UeSho1-e GFP expression strain was constructed,and it was found that the UeSho1 protein was localized in the cell membrane.The expression analysis of UeSho1 revealed that it was in duced to express during the fusion and infection.Ue T14△UeSho1 and Ue T55△UeSho1 were obtained by PEG mediated protoplast transformation method.The study found that the deletion of UeSho1 had no effect on the morphology and growth rate of haploid strains,the formation of conjugation tube and hyphae,the growth of hyphae.However,it affected the formation of fusion of white aerial hyphae and teliospores formation in Zizania latifoli swollen stems.The UeMsb2 was cloned.The length of UeMsb2 c DNA is 3015 bp,without intron,encoding 1004 amino acids,containing Serine-rich domain.The UeMsb2-e GFP expression strain was constructed,and it was found that the UeMsb2 protein was localized in the Vacuole.The expression analysis of UeMsb2 revealed that it was in duced to express during the haploid budding,fusion and infection.Ue T14△UeMsb2 and Ue T55△UeMsb2 strains were obtained by PEG-mediated transformation of protoplasts.The study found that the deletion of UeMsb2 had no effect on the morphology and growth rate of haploid strains,the formation of conjugation tube and pathogenicity,but affected the length and number of hyphae.Ue T14△UeMsb2△UeSho1 and Ue T55△UeMsb2△UeSho1 strains,double deletion UeSho1 and UeMsb2,were obtained by PEG-mediated transformation of protoplasts.We found that double deletion strains were similar to the deletion of UeSho1 strain in haploid budding,vitro fusion and pathogenicity.In addition,we also found that UeSho1 and UeMsb2 were almost not involved in the oxidation,cell wall integrity(CWI)or temperature stress response at the haploid budding stage,at the functions were not redundant.However,UeSho1 and UeMsb2 genes have redundancy in regulating high osmosis stress during budding.Further,the study showed that the deletion of UeSho1 and UeMsb2 had significant effects on the downstream gene expression by q RT-PCR,which were related to the proliferation,filamentous growth and pathogenic of Ustilago esculenta.