基于卤虫致死试验分离淡色生赤壳菌YLZ42杀虫活性代谢产物  

Isolation of an insecticidal metabolite from Bionectria ochroleuca YLZ42 based on the brine shrimp lethality test

作　　者：曾丹 钟娟[1] 谭红[1] 周金燕[1,2] ZENG Dan;ZHONG Juan;TAN Hong;ZHOU Jinyan(Key Laboratory of Envtronmental and Apphed Microbtology/Chengdu Institute of Biology,Chinese Academy of Sciences,Chengdu 610041,China;Environmental Microbiology Key Laboratory of Sichuan Province,Chengdu 610041,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院成都生物研究所/环境与应用微生物重点实验室,成都610041 [2]中国科学院大学,成都610041 [3]四川省环境微生物重点实验室,北京100049

出　　处：《应用与环境生物学报》2018年第5期1009-1014,共6页Chinese Journal of Applied and Environmental Biology

基　　金：四川省科技支撑计划项目(2016NZ0032)资助~~

摘　　要：淡色生赤壳菌具有优良的生防潜力.本实验室前期从广西玉林土壤中分离到一株对小菜蛾等多种农业害虫具有毒杀作用的淡色生赤壳菌YLZ42.采用杀虫活性追踪的方法,用改进后的卤虫致死试验（Brine shrimp lethality test,BSLT）进行活性评价,通过硅胶柱层析、萃取、薄层制备、液相制备的手段进行分离纯化,并探索活性化合物的生物活性和稳定性.分离到杀虫活性化合物纯品,高分辨率质谱显示该活性化合物相对分子质量为788,其对卤虫24 h的LC50为86.89 mg/L.该化合物在温度低于30℃,pH 6-8以及光照和紫外光照条件下活性稳定.该杀虫活性化合物的氢谱与已报道过的分子量为788的化合物均不相同,有望开发为新的微生物源杀虫剂.Bionectria ochroleuca is potentially useful for bio-control. The strain B. ochroleuca YLZ42 isolated from the soil in Yulin of the Guangxi Zhuang Autonomous Region of China was previously shown to be able to kill various agricultural pests, including the diamondback moth, Plutella xylostella. This study aimed to isolate the insecticidal metabolite from the fermentation broth of B. ochroleuca YLZ42 and study its biological activity and stability. The isolation of the insecticidal metabolite was based on a bioassay-guided method. As a consequence, the optimized brine shrimp lethality test （BSLT） was chosen to evaluate the metabolite＇s insecticidal ability. Silica gel column chromatography, extraction, preparative thin layer chromatography （TLC）, and reverse-phase high-performance liquid chromatography （HPLC） were conducted to isolate the insecticidal metabolite. After isolation, the biological activity and stability of this insecticidal metabolite were then studied. Furthermore, high-resolution electrospray ionization mass spectrometry （HR-ESI-MS） and proton nuclear magnetic resonance （1H-NMR） were used to further analyze the metabolite. A pure metabolite with insecticidal activity was obtained. The lethal concentration （LC50） of this metabolite against brine shrimp was found to be 86.89 mg/L after 24 h post-treatment. In addition, the stability of this insecticidal metabolite indicated that its insecticidal activity was stable while the temperature was below 30 ℃ and the pH was between 6 and 8. Further, the metabolite was not sensitive to light in the UV to visible range. The molecular weight of this compound was determined with HR-ESI-MS to be 788. The 1H-NMR spectrum of this insecticidal metabolite was different from that of any other reported compound with the same molecular weight. The insecticidal metabolite has the potential to be developed as a new microbe-derived pesticide.

关 键 词：淡色生赤壳菌 卤虫致死试验 杀虫活性 稳定性 

分 类 号：S476[农业科学—农业昆虫与害虫防治]