控制稻瘟菌菌丝生长基因的克隆-植物病理学专业毕业论文.docxVIP

AbstractMagnaporthe Abstract Magnaporthe grisea,which CaB infect many grasses，is the pathogenic fungus causing rice blast disease．It has been used as a model for studying interactions between plant and phytopathogenic fungi． Isolate$2514 that obtained by screening REMI lransformant library is a slow-growth mutant．On the CM solid medium，hyphal growth rate of$2514 is 45％ofthe wild type strain P131．Also，Isolate S2514 is not only a mutant with slow hyphal growth rate but also with abnormal conidia and pathogenic deficiency．Compared with the wild strain P131．the conidia of S2514 is shorter and wider；and is completely lost its pathogenicity to the susceptible cultivar．The result ofgenetic analysis indicated that the change in hyphal growth，conidia morphology and pathogenicity is caused by the insertion of the plasmid through REMI． Southern blot analysis revealed that the mutant had a single site insertion ofthe marked plasmid in its genome and a 18kb sequence flanking the right side of insertion plasmid was yield．By comparing the flanking sequence with that of the rice blast genome，it was found that the flanking sequence WaS located on contj92．1938 of the Chromosome V．However the flanking sequence on left side of the insertion is located on contil92．610 ofthe Chromosome VII．The leason that two flanking sequences is located on different Chromosomes may be due to DNA deletion or DNA recombination during REMI． GENSCAN with the right side flanking sequence revealed a gene encodes a protein with predicted 656 amino acids Was inactivate．To confirm the gene function，the flanking sequence Was used as a probe to screen the genomic TAC library of the wild type strain Y34，and 9 positive clones were obtained．A fragment containing the whole gene from the positive clone Was cut by Sac I and Xho I and Was cloned into the vector 7afNeu with Neuomycin resistance to transform the mutant．CompIementation test are in process． Key words：Magnaporth grisea，hyphal growth，REMI，plasmid rescue，posit