• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

一株马铃薯干腐病拮抗菌芽孢杆菌的筛选、鉴定及生防因子初探

Screening, Identification, and Preliminary Analysis on Antagonistic Bacillus sp. against Potato Dry Rot

  • 摘要:
      目的  探究分离自青稞白酒糟醅的菌株对马铃薯干腐病的生防效果,为马铃薯干腐病的生物防控提供初步的理论依据。
      方法  采用平板对峙法,测定9株分离自青稞白酒糟醅的菌株对4株马铃薯干腐病病原真菌的抑菌活性。通过形态学、生理生化及分子生物学方法对活性菌株进行鉴定。采用平板透明圈法和排油圈法,分别测定活性菌株发酵液分泌抑菌蛋白和产生脂肽类物质的能力。采用牛津杯法,以马铃薯干腐病病原菌为指示菌,测定活性菌株发酵液有机溶剂萃取物的抑菌活性及其在不同条件下抑菌活性的稳定性。
      结果  (1)筛选到2株对马铃薯干腐病菌具有较强抑制活性的菌株JZ3-4-7和JZ3-1-15,其中JZ3-1-15抑菌活性稳定。(2)经鉴定,JZ3-1-15为贝莱斯芽孢杆菌(Bacillus velezensis)。(3)JZ3-1-15的发酵液能产生蛋白酶、淀粉酶和纤维素酶,并且能够产生脂肽类物质。(4)JZ3-1-15发酵液的正丁醇萃取物对马铃薯干腐病病原真菌青9D-2-6的抑菌活性最高,抑制率达80.07%。(5)JZ3-1-15发酵液的正丁醇萃取物可耐121 ℃高温,热稳定性较强;具有较高的酸碱稳定性,pH值为8时抑菌活性达到最高;紫外线照射处理对抑菌活性无影响。
      结论  JZ3-1-15及其正丁醇萃取物对马铃薯干腐病菌具有强抑菌活性,有望将其开发为马铃薯新型生防保鲜菌剂,并从中分离到对马铃薯干腐病菌具有新颖作用靶点的活性化合物及先导化合物。

     

    Abstract:
      Objective  Bacillus sp. isolated from Hordeum vulgare L. distiller’s grain were explored as a potential biocontrol agent against the potato dry rot disease.
      Method  The plate confrontation method was applied to examine the antagonistic activities of 9 Bacillus strains isolated from the barley wine distiller’s grain against 4 potato dry rot pathogens. Active strains were then identified by morphological, physiological, biochemical, and molecular biology methods. The plate transparent ring and discharge of oil ring tests were used to determine the capacity of the selected strains in secreting bacteriostatic proteins and lipopeptides. Using the potato dry rot pathogens as indicator in the Oxford cup test, the antibacterial activity and the stability under different conditions of the extracts of the fermentation broths of the active strains were determined.
      Result  (1) The Bacillus sp., JZ3-4-7 and JZ3-1-15 displayed significant inhibitory activities against the potato dry rot pathogens in the screening. JZ3-1-15 also exhibited a desirable stability. (2) JZ3-1-15 was subsequently identified as B. velezensis. (3) The JZ3-1-15 fermentation broth contained protease, amylase, cellulase, as well as lipopeptides. (4) The n-butanol extract of JZ3-1-15 fermentation broth had a significant inhibitory rate of 80.07% against the pathogen, Qing 9D-2-6. (5) The same extract withstood a temperature as high as 121 ℃ maintaining a strong thermal stability. It was also stable under acidic and alkaline conditions with a peak anti-pathogenicity at pH 8, and its activity not affected by ultraviolet.
      Conclusion  JZ3-1-15 and the n-butanol extract of its fermentation broth showed significant inhibitory activity on the potato dry rot pathogens. It appeared that they could be developed as a new biocontrol agent against the target pathogens and/or a preservative for fresh potatoes. The effective compounds or precursors might also be isolated from them for combating the potato dry rot disease and further studies.

     

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