稻瘟病菌相关基因camk和chk1的克隆和功能分析-环境科学专业论文.pdf.docxVIP

Ab Ab stract Magnaporthe oryzae is a well-known ascomycetous fungus that causes rice blast· Understanding of the molecular basis of this disease is not only beneficial for rice blast control，but also can serve as a model for revealing other fungal pathogen-plant interactions． Identification of pathogenicity—relative genes is a key step towards this goal．Mutation analysis of pathogen is a powerful strategy in study of the parasitic interaction between the pathogenic fungus and its plant host，which might make the identification of pathogenicity—relative genes possible．In this study,CAMK and CHKl gene from坛oryzae were cloned．The results are showed as follows： 1．CAMK was found from a library of mature appressorium eDNA and its eDNA clones were obtained by PCR strategy,while the sequence homologues were found by using the yeast amino acid sequence of CAMK and CHKl to search from朋·oryzae genome databases，assigned as CAMK and CHKl The genomic DNA clones covered the full length of ORF and eDNA clones were obtained by PCR strategy and the nucleotide sequences of the genomic DNA and eDNA were analyzed． 乞 Hygromycin．resistance transformants were obtained by CAMK and CHKl gene replacement vector construction and fungal transformation．Knockout mutants were identified by PCR and Southern blot．Single copy was detected by genomic Southern blot in the genome of Guyll for each gene． 生 The colony modality of ACAMK was different from Guyll．Compared to Guyll，the growth rate of the mutants on CM，CM．C，CM—N and CM with 1M NaCl had no obvious change．Conidia germination and appressoria formation of mutant ACAMK was different from that of wild type． t Several eGFP fusion expressing vectors，including PHNSCHKlE，PHNECHKl and PHCHKIPE were constructed． 5 The colony modality of ACAMK was different from Guyll．Compared to Guyll，the growth rate of the mutants on CM，CM．C，CM—N and CM with 1M NaCI had no obvious 1I change．Conidia change．Conidia germination and appressoria formation of mutant zl