Molecular genetic analysis of genes involved in melanin biosynthesis in plant pathogenic fungi utilizing its transformation system

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In this research, transformation of a Plant Pathogenic funongi was constructed and a gene concerning melanin biosynthesis, one of the pathogenic factors, of Colletotrichum lagenarium was cloned. The results are as follow.1. Protoplast system of C.lagenarium , which can be used for both isolation of genomic DNA and reception of foreign DNA, was constructed. 2. Benomyl resistant beta-tubulin gene was cloned from benomyl resistant C.lagenarium mutant. The gene was utilized as a selection marker for transformation of this fungus. 3. Cosmid vector for construction of genomic DNA library of C. lagenarium was constructed using resistant beta-tubulin gene as a selection marker. 4. Genomic DNA library of C.lagenarium was constructed in the cosmid vector. 5. Genomic library concerning melanin biosynthesis was screened by transformation of an albino mutant by the genomic library DNA. 6. A gene determining the phenotype of albino mutant was cloned by isolating the cosmid DNA from a melanin restored transformant.