青枯雷尔氏菌拮抗放线菌的筛选及其抗菌活性物质的分离
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摘要
烟草青枯病是由青枯雷尔氏菌引起的土传性病害。从健康土壤中分离到对青枯雷尔氏菌具有明显拮抗作用的放线菌LC-7,经初步鉴定为链霉菌属(Streptomyces sp.)。链霉菌LC-7的发酵液用氯仿、正丁醇萃取后,通过捷克八溶剂系统分析,判定该抗菌物质为碱性水溶性抗生素。抗菌物质进一步采用离子交换树脂纯化,并经醇沉后,获得抗生素的结晶,再用HPLC进一步纯化后并进行质谱分析可得出,LC-7所产抗菌活性物质为一种新的氨基糖苷类抗生素,其分子量为365.1。田间试验表明,链霉菌LC-7对烟草青枯病具有显著的防治效果,防效为85.48%。
Tobacco bacterial wilt is a soil-borne disease caused by Ralstonia solanaceanm. A strain of Actinomycete LC-7 with obvious antagonistic activities against R. solanaceanm was isolated from healthy soils and identified as Streptomyces sp. The fermentation broth of Streptomyces sp. LC-7 was extracted with chloroform and n-butanol. The analysis through the Czech eight solvent systems revealed that the antibacterial substance was an alkaline water-soluble antibiotic. The antibacterial material was purified by exchange column, and precipitated with alcohol to obtain the crystal. The crystal was further purified by HPLC and analyzed by mass spectrometry. The results showed that the antimicrobial active substance was a new aminoglycoside antibiotic with a molecular weight of 365.1. In the field experiment, Streptomyces sp. LC-7 had significant control effect on tobacco bacterial wilt. The control efficacy was 85.48%.
Tobacco bacterial wilt is a soil-borne disease caused by Ralstonia solanaceanm. A strain of Actinomycete LC-7 with obvious antagonistic activities against R. solanaceanm was isolated from healthy soils and identified as Streptomyces sp. The fermentation broth of Streptomyces sp. LC-7 was extracted with chloroform and n-butanol. The analysis through the Czech eight solvent systems revealed that the antibacterial substance was an alkaline water-soluble antibiotic. The antibacterial material was purified by exchange column, and precipitated with alcohol to obtain the crystal. The crystal was further purified by HPLC and analyzed by mass spectrometry. The results showed that the antimicrobial active substance was a new aminoglycoside antibiotic with a molecular weight of 365.1. In the field experiment, Streptomyces sp. LC-7 had significant control effect on tobacco bacterial wilt. The control efficacy was 85.48%.
引文
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[17]陆铮铮,彭丽娟,丁海霞,等.烟草青枯菌拮抗放线菌的筛选及鉴定[J].中国烟草科学,2013,34(2):54-58.
[18]熊仕俊,孙成龙,施闯,等.番茄青枯病菌拮抗放线菌的筛选及鉴定[J].北方园艺,2014(5):114-117.
[19]龙克启,陆铮铮,吴石平,等.一株放线菌对辣椒青枯菌的拮抗作用及其鉴定[J].河南农业科学,2012,41(11):92-95.
[20]王东胜.秦岭主峰太白山北坡5种生境中微生物区系及拮抗放线菌资源研究[D].西安:西北农林科技大学,2014.
[21]李定刚.氨基糖苷类抗生素的研究应用概述[J].北方牧业,2008(2):27-27.
[2]陈程,黎定军,陈武.烟草青枯病生物防治研究进展[J].作物研究,2012,25(6):639-642.
[3]陈勇,金晨钟.烟草青枯病的发生与防治[J].现代农业科技,2009(10):91-93.
[4]番华彩,唐嘉义,秦小萍.烟草青枯病防治研究进展[J].云南大学学报(自然科学版),2008(S1):31-35.
[5]霍沁建,张深,王若焱.烟草青枯病研究进展[J].植物保护科学,2007,23(8):364-368.
[6]黄路枝.代谢农药活性物质稀有放线菌的筛选[D].西安:西北农林科技大学,2007.
[7]LIU Y,SHI J,FENG Y.Tobacco bacterial wilt can be biologically controlled by the application of antagonistic strains in combination with organic fertilizer[J].Biology and fertility of soils,2013,49(4):447-464.
[8]孙立广,张洪春,赵秀云,等.烟草青枯病拮抗菌在有机肥中的定殖效率及田间防治效果[J].中国烟草科学,2016,37(4):48-53.
[9]安德荣,慕小倩,刘翠娟,等.土壤拮抗放线菌的分离和筛选[J].微生物学杂志,2002(5):1-3.
[10]李毅,王国平,张克诚.拮抗放线菌LJ50和MJ52的分离与初步鉴定[J].湖南农业大学学报,2005(3):310-313.
[11]周德庆.微生物实验手册[M].上海:上海科学技术出版社,1997,339-345.
[12]陈丽琼,郭永夏,马国秋.抗生素4-215的分离、提取和鉴定[J].微生物学报,1980,20(1):94-99
[13]陈丽,疏秀林,安德荣,等.土壤拮抗放线菌S-5210-6活性产物抑菌谱及其作用机制[J].植物保护学报,2007(3):277-282.
[14]TAN H M,ZHOU S N,DENG Z J,et al.Ribosomalsequence-directed selection for endophytic streptomycete strains antagonistic to Ralstonia solanacearum to control tomato bacterial wilt[J].Biological Control 2011,59:245-254.
[15]梁亚萍,宗兆锋,马强.6株野生植物内生放线菌防病促生作用的初步研究[J].西北农林科技大学学报,2007,35(7):131-136.
[16]蒋承耿,刘琼,陈鹏.烟草青枯病防治药剂室内筛选及田间药效研究[J].安徽农业科学,2013(3):1098-1099.
[17]陆铮铮,彭丽娟,丁海霞,等.烟草青枯菌拮抗放线菌的筛选及鉴定[J].中国烟草科学,2013,34(2):54-58.
[18]熊仕俊,孙成龙,施闯,等.番茄青枯病菌拮抗放线菌的筛选及鉴定[J].北方园艺,2014(5):114-117.
[19]龙克启,陆铮铮,吴石平,等.一株放线菌对辣椒青枯菌的拮抗作用及其鉴定[J].河南农业科学,2012,41(11):92-95.
[20]王东胜.秦岭主峰太白山北坡5种生境中微生物区系及拮抗放线菌资源研究[D].西安:西北农林科技大学,2014.
[21]李定刚.氨基糖苷类抗生素的研究应用概述[J].北方牧业,2008(2):27-27.