Abstract
Treatment with cyclic AMP (cAMP) induces appressorium formation in the phytopathogenic fungus Magnaporthe grisea, the causative agent of rice blast disease. In a search for the M. grisea genes responsible for appressorium formation and host invasion, SAGE (Serial Analysis of Gene Expression) was carried out using mRNA isolated from fungal conidia germinating in the presence and absence of cAMP. From cAMP-treated conidia 5087 tags including 2889 unique tags were isolated, whereas untreated conidia yielded 2342 unique tags out of total of 3938. cAMP treatment resulted in up- and down-regulation of genes corresponding to 57 and 53 unique tags, respectively. Upon consultation of EST/cDNA databases, 22 tags with higher representation in cAMP-treated conidia were annotated with putative gene names. Furthermore, 28 tags corresponding to cAMP-induced genes could be annotated with the help of the recently published genome sequence of M. grisea. cAMP-induced genes identified by SAGE included many genes that have not been described so far, as well as a number of genes known to be involved in pathogenicity, e.g. MPG1, MAS1 and MAC1. RT-PCR of 13 randomly selected genes confirmed the SAGE results, verifying the fidelity of the SAGE data.
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Acknowledgements
We thank Dr. K. Kinzler (Johns Hopkins University, Baltimore) for provision of the SAGE protocol and SAGE analysis software, Dr. S. Koizumi (Tohoku Agricultural Research Center, MAFF, Japan) for the gift of M. grisea INE86-137, and Dr. N. Talbot (University of Exeter, Exeter, UK) for invaluable discussion. We are grateful to Dr. Y. Takano (Kyoto University, Kyoto, Japan) for invaluable suggestions and for technical advice concerning fungal cultures and RNA isolation from M. grisea. Thanks are due to Dr. P. C. Sharma (Ch. Charan Singh University, Meerut, India) and to Dr. S. Rakshit (Directorate of Maize Research, Indian Agricultural Research Institute, India) for the improvement of the manuscript. This work was partly supported by Grant No. S1-1105 from the Institute of Bioresources, Tsukuba, Japan, and by the Research for Future Program of the Japan Society for the Promotion of Science
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Irie, T., Matsumura, H., Terauchi, R. et al. Serial Analysis of Gene Expression (SAGE) of Magnaporthe grisea: genes involved in appressorium formation. Mol Genet Genomics 270, 181–189 (2003). https://doi.org/10.1007/s00438-003-0911-6
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DOI: https://doi.org/10.1007/s00438-003-0911-6