| Phytophthora infestans is a kind of plant pathogens causing potato and tomato late blight.Its sexual reproduction plays an important role in the evolution and continuation of this species.However,up to now,there have been few studies focused on the molecular mechanism of this biological process.In this study,the proteome of P.infestanse respectively treated with hormone al and DMSO was determined by iTRAQ technology with the strain P7723 as the material.Then,two differentially expressing genes,PITG-08690T0 and PITG-09593T0,were selected to study their functions.The expression profile of the two genes at different development stages was measured by real time R.T-PCR technology.The subcelluar localization of the proteins was detected by fluorescent protein fused expression technology.One way anova analysis of the protomics data showed that there were 795 differentially expressed proteins under hormone al treatment and 594 of them existed only in the hormone αl treatment group compared with that in the DMSO control group.Throuth GO annotation at level 2,7 differential proteins existed only or were expressed at a high level in the hormone al treatment group.T test analysis of the data discovered 183 differentially expressed proteins,of which there were 10 down and 25 up regulated proteins at 3h,10 down and 4 up regulated proteins at 8h,12 down and 5 up regulated proteins at 24h,and 60 down and 78 up regulated proteins at 72h after the hormone α1 treatment.The enrichment analysis showed there were totally 10 down-regulated and 13 up-regulated proteins.The sporangium amount of the strain P7723.growing on the CA medium with 40 ℃ treatment was the most,reaching 767±7.98 个/cm2.The best condition preparing the protoplasts was to add 30mL of 10 mg/mL Driselase and Lysing Enzymes into the mycelia from 400mL liquid medium and inoculated at room temperature for 90 min.The coding regions of PITG-08690T0 and PITG-09593T0 cDNA were 1233bp and 501bp respectively and the amino acid sequences of the two proteins deduced according to the cDNA were the same as that published on NCBI web.Both the expression level of the gene PITG-08690T0 and PITG-09593T0 were the highest in the cysts and decreased gradually in zoospore,germinating cysts,sporangia,and mycelia of the strain P7723.The PITG-08690T0 protein may located in cytoplasm.The differentially expressed proteins of P.infestans related with its sexual reproduction were revealed and the functions of two genes,PITG-08690 and PITG-09593,were researched preliminarily in this study,which laid a foundation for the discovery of the signal pathway of sexual reproduction in P.infestanse and for the effective control of plant diseases caused by this pathogen. |
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