呼吸抑制剂对烟草灰霉病的防治及其作用机理的研究

Gray mold of tobacco (Nicotiana tabacum L.) caused by Botrytis cinerea Pers.:Fr is one of the important tobacco diseases worldwide. It could cause serious damages during seedling period in the nursery bed and also on tobacco leaves in the field. More attention should be paid to solve the problem. In recent years, fungicides affecting fungal respiration, such as inhibitors of mitochondrial complex II boscalid and complex III azoxystrobin, uncoupler fluazinam and multi-site toxicant mancozeb, become the major botryticides for gray mold management in many crops. However, to the best of our knowledge, they have not been used in flue-cured tobacco for gray mold management in China. Additionally, the differences in mode of action of the four kinds of inhibitors are still unclear. Phenomics is a novel technology with typical advantages in high throughput, and is widely used in microorganism researches. The objective of this study is to assess the four kinds of respiration inhititors for gray mold management in tobacco; to test the metabolic phenotypes of B. cinerea, especially the metabolic phenotypes of the pathogen when inhibited, respectively, by the four kinds of respiration inhibitors; and to analyze the differences in mode of action of the four kinds of respiration inhibitors through metabolic phenomics. Results of this study will explain the modes of action of these respiration inhibitors from phenomics, and will provide useful information for tobacco gray mold management.

灰葡萄孢引起的烟草灰霉病是国内外重要的烟草病害之一，在漂浮育苗期和大田期造成危害，严重威胁烟草的生产，亟待解决。近年来，线粒体电子传递链复合物Ⅱ抑制剂啶酰菌胺、复合物Ⅲ抑制剂嘧菌酯、解偶联剂氟啶胺及多作用位点化学杀菌剂代森锰锌逐渐成为灰霉病防治的主要药剂，但还未在我国烟草灰霉病的防治上进行应用，它们在作用机理上的差异仍不完全清楚。本申请拟开展4类呼吸抑制剂防治烟草灰霉病的应用技术研究；同时拟采用微生物领域具有独特优势的高通量表型芯片技术，开展烟草灰霉病菌的表型组学分析；分别开展线粒体电子传递链复合物Ⅱ抑剂啶酰菌胺、复合物Ⅲ抑制剂嘧菌酯、解偶联剂氟啶胺及多作用位点化学杀菌剂代森锰锌的作用下，烟草灰霉病菌的代谢表型分析，从灰霉病菌代谢表型上揭示4类呼吸抑制剂在作用机理上的差异。该项目拟从代谢表型水平解释4类呼吸抑制剂的作用机理，对指导烟草灰霉病的化学防治具有重要的理论意义。

灰葡萄孢引起的烟草灰霉病是国内外重要的烟草病害之一，在漂浮育苗期和大田期造成危害，严重威胁着烟草的生产，亟待解决。近年来，线粒体电子传递链抑制剂逐渐成为灰霉病防治的主要药剂，但还未在我国烟草灰霉病的防治上进行应用，它们在作用机理上的差异仍不完全清楚。本项目开展了4类呼吸抑制剂防治烟草灰霉病的应用技术研究；同时采用目前微生物领域广泛使用和具有独特优势的高通量表型芯片技术，开展了烟草灰霉病菌的表型组学分析；分别开展线粒体电子传递链复合物Ⅱ抑剂啶酰菌胺、复合物Ⅲ抑制剂嘧菌酯、解偶联剂氟啶胺及多作用位点化学杀菌剂代森锰锌的作用下，烟草灰霉病菌的代谢表型分析。结果表明，嘧菌酯、啶酰菌胺、氟啶胺及代森锰锌对烟草灰霉病菌的菌丝生长和分生孢子萌发均表现出不同的抑菌活性，对烟草灰霉病均有较好的预防效果，嘧菌酯、啶酰菌胺及氟啶胺还具有较好的治疗效果。烟草灰霉病菌能代谢13%的碳源、82%的氮源、100%的磷源和硫源，具有95种生物合成途径、81种渗透压代谢表型、43种pH代谢表型和脱氨酶活性，不具脱羧酶活性。基于菌丝代谢表型的作用机理研究发现，呼吸抑制剂嘧菌酯主要抑制灰霉病菌的三羧酸循环（TCA）、糖酵解、尿素循环、嘌呤代谢及嘧啶代谢；氟啶胺和啶酰菌胺主要抑制灰霉病菌的TCA、糖酵解及半乳糖代谢；代森锰锌主要抑制其TCA、糖酵解、尿素循环及嘌呤代谢。基于孢子萌发代谢表型的作用机理研究发现，呼吸抑制剂嘧菌酯主要抑制灰霉病菌的TCA、糖酵解、烟酰胺腺嘌呤二核苷酸（NAD）代谢、尿素循环、嘌呤代谢及尿素循环；氟啶胺主要抑制灰霉病菌的TCA、嘌呤代谢及NAD代谢；啶酰菌胺主要抑制其糖酵解；代森锰锌主要抑制其TCA和糖酵解。研究结果对指导烟草灰霉病的化学防治具有重要意义，同时从代谢表型上揭示了4类呼吸抑制剂在作用机理上的差异。