科研之友
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摘要
To date, a number of genes that are expressed in the early stages of infection have been reported, while few genes that are expressed during the course of colonization, after the initiation of plant infection, have been studied. Plant inoculations, real-time polymerase chain reaction (PCR), gene cloning, protein expression and bioinformatics analysis were combined to identify a novel gene, MgNIP04, in the rice blast fungus, Magnaporthe grisea. Bioinformatics analysis showed that the amino acid sequence contained a signal peptide. The wounded inoculation resulted in necrosis specks when the total expressed proteins including MBP-MgNIP04 was inoculated on the wounded rice leaves, which demonstrated the protein directly interacted with rice. The real-time PCR result of infected leaves showed that it is upregulated during late stages of infection of rice. Additionally, the copies of the gene expression absolute quantity of the gene were 7.61 x 10(2), 1.06 x 10(3), 1.31 x 10(3), 4.13 x 10(3), and 4.00 x 10(3), at 24, 48, 72, 96 and 168 h postinoculation (HPI), respectively. The higher copies of MgNIP04 were found at 96 and 168 HPI. The copies of MgNIP04 in mycelia of Y98-63C, Y99-16, 94-64-1b and 95-23-4a were significantly lower than those of infected leaves. Through the above bioinformatics and experimental analysis, our result suggested that the MgNIP04 was novel pathogenicity-related protein in rice blast fungus.
