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微生物学通报

一株北里孢菌株的分离鉴定及其对松材线虫的致病性
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国家自然科学基金项目(No. 31100104);河南省教育厅科学技术研究重点项目基础研究计划项目(No. 13A180809);河南省高等学校《生物工程专业综合改革试点》项目(教高【2012】859号-41);河南省高等学校《生物科学专业核心课程教学团队》项目(教高【2012】1099号-19);南阳师范学院青年基金项目(QN. 2011002)


Screening, identification and the virulence factor of an actinomycete strain Kitasatospora sp. C620 to Bursaphelenchus xylophilus
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    摘要:

    【目的】筛选、鉴定出对松材线虫杀灭活性较高的放线菌菌株,并确定生防菌株的毒力因子。【方法】采用平板活性测试及代谢杀虫活性检测方法进行筛选,采用形态学及16S rDNA序列分析等进行鉴定。对发酵液中的活性物质稳定性分析后,利用醇沉、萃取、层析、气相色谱/质谱分析等方法分离纯化出杀虫毒力因子。【结果】从河南南阳宝天曼的腐木及枯枝落叶样品中共分离获得了79株放线菌,从中筛选出对松材线虫有灭活作用的放线菌6株,其中分离株C620菌株对松材线虫的灭活性最高:该菌株的发酵液处理松材线虫48、60 h后线虫的死亡率分别达到60.0%、81.5%。结合该菌株的形态学、生理学特征及16S rDNA序列分析等结果将其归为北里孢菌属中的一个种,菌株编号Kitasatospora sp. strain C620。该菌株的发酵液中杀线虫活性物质的热稳定性、光稳定性及耐储藏性均较强,在中性偏碱性环境较稳定;经pH纸电泳层析初步确定该物质属于碱性水溶性物质。对菌株C620发酵液分离纯化,得到活性化合物为1-苯基-3-(2-吡啶)-5-吡唑啉酮。【结论】获得一株松材线虫高效生防菌Kitasatospora sp. strain C620,其活性物质为1-苯基-3-(2-吡啶)-5-吡唑啉酮。

    Abstract:

    [Objective] The actinomycete strains with high nematicidal activity against Bursaphelenchus xylophilus were isolated and identified. The virulence factors were determined. [Methods] Strains were selected by plate activity test and insecticidal activity detection by metabolites. Strains were identified by phenotypic and genotypic characteristics. After analyzing the primary characteristics of the activity substances in the fermented broth, the virulence factor was purified and identified using alcohol precipitation, extracted by chloroform, chromatographer, and GC-MS. [Results] Seventy nine actinomycetes were isolated and six strains which had nematicidal activity to B. xylophilus were totally found from the rotten wood, dry branches and fallen leaves in Baotianman Natural Reserves in Nanyang, Henan Province, China. Among which, an actinomycete strain C620, showed the highest nematicidal activity to B. xylophilus. The culture supernatant from the strain C620 separately killed 60.0% and 81.5% of B. xylophilus within 48 h and 60 h, which suggested the high toxic activity toward the nematodes. The actinomycete was classified to be Kitasatospora genus. The virulence factor in the fermented broth of C620 showed strong thermal stability, light stability and storage resistance. The bioactive substance was stable in alkaline environments and can be dissolved in water. Finally, the compound was found to be 1-phenyl-3-(2-pyridyl)-5-pyrazolone. [Conclusion] An actinomycete strain Kitasatospora sp. C620 with high killing activity against B. xylophilus was isolated and the virulence factor was 1-phenyl-3-(2-pyridyl)-5-pyrazolone.

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王云,郑豪盈,樊永欣,杨建伟,张林,张成,陈慧茹,陈诚,牛秋红. 一株北里孢菌株的分离鉴定及其对松材线虫的致病性[J]. 微生物学通报, 2014, 41(10): 2035-2042

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  • 在线发布日期: 2014-10-10
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