Studies on Oryzalide A, a novel antibacterial diterpene, isolated from healthy leaves of rice plant

从健康水稻叶片中分离出的一种新型抗菌二萜 Oryzalide A 的研究

• 批准号: 02660045
• 负责人: WATANABE Minoru
• 金额: $ 1.28万
• 依托单位: Tokyo University of Agriculture and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02660045/
• 关键词: Rice plant; Bacterial leaf blight; Antibacterial substance; Disease resistance; Oryzalide; Oryzalic acid; Compound A-F; Microanalysis

1. Isolation and identification of ten kinds of Oryzalides and related compounds.A novel antibacterial diterpene substances, Oryzalide A and structurally related 9 substances, were isolated from healthy rice leaves and the structures were determined. An ethyl acetate-extractable acidic and neutral crude extracts were isolated from about 20 kg fresh weight of flag leaves of cv. Norin 27, and they were purified by silica gel and sephadex LH-20 column chromatography, HPLC, partition TLC, etc., and finally their structures were determined by ^1H-NMR, ^<13>C-NMR, HR-MS, IR, etc. As the results, four kinds of acidic substances of Oryzalides A, B, and Oryzalic acids A, B, and six kinds of neutral substances of Compounds A, B, C, D, E and F were newly found. The structural characteristics, yields and antibacterial activities of the above mentioned ten substances were clarified.2. Establishment of a microanalytical method of Oryzalides and Oryzalic acids, and measurement of an endogenous level … More in rice leaves. Oryzalides(A and B)and Oryzalic acids(A and B)were analyzed by means of GC-MS-SIM equipped with a capillary column after conversions to the Methyl ester derivative. Amounts of endogenous Oiyzalides and Oryzalic acids were determined by addition of deuterated labeled[17-D_2]Oryzalide B as an internal standard. Major components were found to be Oryzalide A and Oryzalic acid A in fresh leaves of rice plant when monitoring at m/z 305 fragment ion, which was a characteristic and common ion among the four substances.As the results of preliminary measurement using the above microanalytical method, all Oryzalides and Oryzalic acids were detected from healthy leaves of 8 rice cultivars different in their resistance to bacterial leaf blight, but no correlation was observed between the amount and the degree of resistance. An endogenous four substances were almost detected from leaves but not from roots and ears, and their amount increased as a growth of rice plant proceeds. When the leaves were wounded by a needle bundle-prick, the amount increased to 3-4 times compared with that of not wounding healthy leaves(control). In the leaves inoculated with a compatible or an incompatible strain of pathogenic bacterium, the amount increased to 7.2 or 9.1 times of control, respectively. Less

1. 10种Oryzalides及相关化合物的分离鉴定。从健康稻叶中分离得到一种新型抗菌二萜类物质Oryzalide A及结构相关的9个物质，并测定了乙酸乙酯可提取的酸性和中性粗提物的结构。从大约20公斤鲜重的cv Norin 27旗叶中分离出它们，并通过硅胶和葡聚糖凝胶纯化。 LH-20柱层析、HPLC、分配TLC等，最后通过 1 H-NMR、 13 C-NMR、HR-MS、IR等确定其结构。结果，四种酸性化合物新发现了Oryzalides A、B和Oryzalic Acids A、B物质，以及化合物A、B、C、D、E、F中六种中性物质的结构特征、收率和结果。阐明了上述10种物质的抗菌活性。 2.稻叶中稻内酯类和稻谷酸的微量分析方法的建立及稻叶中稻谷内酯类(A和B)和稻谷酸(A和B)的内源水平测定。转化为甲酯衍生物后，通过配有毛细管柱的 GC-MS-SIM 进行分析。以氘代标记的[17-D_2]Oryzalide B为内标测定Oyzalides和Oryzalic Acids，在m/z 305碎片离子下监测，发现水稻新鲜叶片中的主要成分为Oryzalide A和Oryzalic Acid A。 ，是这四种物质中的特征离子和共有离子。使用上述微量分析方法进行初步测量的结果，检测到了所有的Oryzalides和Oryzalic Acids 8个水稻品种的健康叶片中对白叶枯病的抗性不同，但含量与抗性程度之间没有相关性。内源四种物质几乎在叶片中检出，但在根和穗中未检出，且含量有所增加。随着水稻生长的进行，用针束刺伤叶片时，接种相容剂或不相容剂的叶片中，其数量比未伤及健康叶片的叶片增加3-4倍。病原体菌株细菌，数量分别增加至对照的7.2或9.1倍。

项目成果

Structures of Oryzalides A and B, and Oryzalic Acid A, a Group of Novel Antimicrobial Diterpenes, Isolated from Healthy Leaves of a Bacterial Leaf Blight-resistant Cultivar of Rice Plant

从抗细菌叶枯病水稻品种健康叶片中分离出的一组新型抗菌二萜 Oryzalides A 和 B 以及 Oryzalic Acid A 的结构

• DOI: 10.1271/bbb1961.55.803
• 发表时间: 1991-03-01
• 期刊: Agricultural and biological chemistry
• 作者: Y. Kono;Jim Uzawa;Kimiko Kobayashi;Yoshikatsu Suzuki;M. Uramoto;A. Sakurai;Minoru Watanabe;T. Teraoka;D. Hosokawa;Manabu Watanabe;Hideaki Kondo
• 通讯作者: Hideaki Kondo