A study on the role of catalase from Magnaporthe grisea and an inhibitor of catalase from rice cells in the rice-blast
稻瘟病菌过氧化氢酶和水稻细胞过氧化氢酶抑制剂在稻瘟病中作用的研究
基本信息
- 批准号:16580033
- 负责人:
- 金额:$ 2.5万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2004
- 资助国家:日本
- 起止时间:2004 至 2006
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
In order to develop a novel experimental system to analyze the molecular mode of interaction between rice cells (Oryza sativa cv. Nipponbare) and rice blast fungus (Magnaporthe grisea), suspension-cultured cells of rice were co-cultured with conidia suspension of various field isolates of the rice blast fungus and analyzed defense responses by rice including the production of H_2O_2 and expression of defense-related genes. The level of H_2O_2 was higher when co-cultured with incompatible fungal isolates than with compatible ones. The difference was due not to the gene-for-gene relationship but to the activity of catalase found in the conidia suspension, the level of which was specific to each isolates. In the leaf sheath assay for up to 48 h, inoculation with both compatible and incompatible spores washed to remove catalase activity resulted in a significant reduction in invasion. The addition of catalase to the washed spore suspension increased the efficiency of invasion. The removal or addition of catalase in the spore suspension was related to the level of accumulated H_2O_2 in the infected leaf cells as observed by staining with diaminobenthidine. In the leaf blade assay, inoculation of washed compatible spores formed smaller lesions than washed spores with added catalase at 5 days. These results suggested that fungal catalase is a pathogenesis-related factor in the rice blast fungus. Similar catalase activities were detected in the culture filtrate of the fungus, and the enzyme was partially purified from the culture filtrate by gel filtration, DEAE-cellulose column chromatography and SDS-polyacrylamide gel electrophoresis. Analysis of the endogenous amino acid sequences after digestion of the protein with protease showed that the H_2O_2-degrading activity is carried by a catalase/peroxidase that is specifically found in eukaryotic microbes. In Magnaporthe grisea, a single-copied gene encodes the enzyme.
为了建立一种新的水稻细胞间相互作用的分子模式分析系统,对水稻细胞间相互作用的分子机制进行了研究。以水稻悬浮培养细胞为材料,与稻瘟病菌不同田间分离物的分生孢子悬浮液共培养,研究了水稻对H_2O_2的产生和防御相关基因的表达。与不亲和菌株共培养时,H_2O_2水平高于与亲和菌株共培养。这种差异不是由于基因对基因的关系,而是由于在分生孢子悬浮液中发现的过氧化氢酶的活性,其水平是特定于每个菌株。在叶鞘测定长达48小时,接种与兼容和不兼容的孢子洗涤去除过氧化氢酶活性导致入侵显着减少。在洗涤过的孢子悬浮液中加入过氧化氢酶可提高入侵效率。用二氨基苄噻啶染色观察到,孢子悬浮液中过氧化氢酶的去除或添加与侵染叶细胞中积累的H_2O_2水平有关。在叶片测定中,在第5天,接种洗涤的相容孢子形成比添加过氧化氢酶的洗涤孢子更小的病变。这些结果表明,真菌过氧化氢酶是稻瘟病菌致病相关因子。在该菌的培养滤液中检测到类似的过氧化氢酶活性,并通过凝胶过滤、DEAE-纤维素柱层析和SDS-聚丙烯酰胺凝胶电泳从培养滤液中部分纯化了该酶。经蛋白酶酶解后,对内源氨基酸序列的分析表明,该蛋白具有过氧化氢酶/过氧化物酶活性,该酶是真核微生物特有的。在稻瘟菌中,一个单拷贝基因编码该酶。
项目成果
期刊论文数量(2)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Elicitor and calatalse activity of conidia suspensions of various strains of Magnaporthe grisea in suspension-cultured cells of rice
- DOI:10.1271/bbb.70684
- 发表时间:2008-03-01
- 期刊:
- 影响因子:1.6
- 作者:Tanabe, Shigeru;Hayashi, Nagao;Minami, Eiichi
- 通讯作者:Minami, Eiichi
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