Development of trunk injection agents for a pine-wilt desease

松材线虫病树干注射剂的研制

• 批准号: 03556016
• 负责人: KAWAZU Kazuyoshi
• 金额: $ 5.31万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03556016/
• 关键词: Nematicide; Bursaphelenchus xylophilus; Paired-ion chromatography; Dehydrosinefungin; nucleoside antibiotic; trunk injection

Dehydrosinefungin is the potent nematicidal compound, but the productivity of this compound by Strepromyces sp. H-114 is very low. Thus, an improvement of the productivity is needed. We developed the quantitative method for dehydrosinefungin to find the best conditions for production of this compound. Paired-ion chromatography using an HPLC aparatus resulted inthe best separation of dehydrosinefungin and its analogs, sinefungin and amidinosinefungin. Guided by the productivity of dehydrosinefungin, we determined the best culture conditions. Using the best conditions, the productivity reached more than 20 mg/l, that is, ten times as high as that of original conditions. Large scale cultivation using a jar-fermentor gave us more than 200 mg of pure dehydrosinefugin. This enabled us to carry out the assay using three-year pine trees.On the other hand, we established the method for a trunk injection of the active compound using morantel tartarate as a test material. Using this method, dehydrosinefungin was injected to three-year pine trees before the treatment of nematodes. We found that 0.5 mg dehydrosinefungin per a three-year pine tree resulted in the resistance of tested trees against wilting. This result indicated that dehydrosinefungin was the promising agent for a pine-wilt desease.

脱氢sinefungin是有效的杀线虫化合物，但链霉菌属H-114对该化合物的产率非常低。因此，需要提高生产率。我们建立了脱氢sinefungin的定量方法，以寻找生产该化合物的最佳条件。采用高效液相色谱仪对离子色谱法分离脱氢sinefungin及其类似物sinefungin和amidinosinefungin效果最好。以脱氢sinefungin产量为指标，确定了最佳培养条件。在最佳条件下，产率可达20 mg/l以上，是原条件下的10倍。使用罐式发酵罐的大规模培养给我们提供了超过200 mg的纯脱氢sinefugin。这使得我们能够使用三年生松树进行测定。另一方面，我们建立了使用酒石酸莫仑太尔作为试验材料的活性化合物树干注射方法。使用这种方法，脱氢sinefungin注射到三年松树线虫治疗前。我们发现，每棵三年生松树0.5毫克脱氢sinefungin导致测试树木对萎蔫的抗性。这一结果表明，脱氢sinefungin是一种很有前途的防治松树枯萎病的药剂。