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

利用转化系统对植物病原真菌黑色素生物合成相关基因进行分子遗传学分析

• 批准号: 63560044
• 负责人: KUBO Yasuyuki
• 金额: $ 1.79万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63560044/
• 关键词: Pathogenicity; Melanin; Plant Pathogenic Fungi; Molecular Genetics; Transformation; 遺伝子解析

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.

本研究构建了植物病原真菌的转化，并克隆了与黑炭疽病菌致病因子之一黑色素生物合成有关的基因。结果如下： 1．构建了可用于基因组DNA分离和外源DNA接收的C.lagenarium原生质体系统。 2.从苯菌灵抗性C.lagenarium突变体中克隆了苯菌灵抗性β-微管蛋白基因。该基因被用作该真菌转化的选择标记。 3.以抗性β-微管蛋白基因为选择标记，构建了用于构建C.lagenarium基因组DNA文库的粘粒载体。 4.在粘粒载体中构建C.lagenarium基因组DNA文库。 5.通过用基因组文库DNA转化白化突变体来筛选与黑色素生物合成有关的基因组文库。 6.通过从黑色素恢复的转化体中分离粘粒DNA来克隆决定白化突变体表型的基因。

项目成果

Yasuyuki Kubo: "The Fungal Spore and Disease Initiation in Plants and Animals" Plenum Publishing Corporation, (1990)

Yasuyuki Kubo：“植物和动物中的真菌孢子和疾病起始”Plenum Publishing Corporation，（1990）

Tanaka, C., Kubo, Y., Tsuda, M.: "Genetic analysis and characterization of Cochliobolus heterostrophus color mutants" Mycological Research, 1990.

Tanaka, C.、Kubo, Y.、Tsuda, M.：“异旋旋毛菌颜色突变体的遗传分析和表征”真菌学研究，1990。

Kubo,Y.: Expenmental Mycology. 13. (1989)

Kubo，Y.：实验真菌学。

Kubo, Y. and Furusawa, I.: "The Fungal Spore and Disease Initiation in Plants and Animals" Plenum Publishing Corporation, 1990.

Kubo, Y. 和 Furusawa, I.：“植物和动物中的真菌孢子和疾病起始”Plenum Publishing Corporation，1990 年。

Tajima,S.: Experimental Mycology. 13. (1989)

Tajima，S.：实验真菌学。