解淀粉芽孢杆菌B10-26抑菌物质的稳定性研究与初步分离纯化

Stability Determination and Prefractionation of Antifungal Substance Produced by Bacillus amyloliquefaciens B10-26

为了将解淀粉芽孢杆菌B10-26应用于植物病害的生物防治,以芝麻茎点枯病菌等13种病原菌为指示菌,采用含毒介质法检测生防菌发酵液的活性;对抑菌物质粗提物在不同温度和pH值条件下处理,采用牛津杯法检测活性;采用高效液相法对抑菌物质进行初步的分离纯化.结果表明,解淀粉芽孢杆菌B10-26发酵液对13种植物病原真菌均有不同程度的抑制作用,对芝麻茎点枯病菌的抑菌率最高,达到91.67％.抑菌物质活性虽然随着温度的升高有所下降,但在121℃高温处理下,仍然产生11 mm的抑菌圈,说明其具有较高的热稳定性;在pH值2～10时,押菌活性没有明显变化,说明其具有较好的酸碱稳定性,但在碱性条件下活性较高.对抑菌物质的初步分离表明,B10-26发酵液中含有多种活性成分.

In order to apply Bacillus amyloliquefaciens B10-26 in the biocontrol of plant diseases,toxic media method was used to measure its inhibitory spectrum on 13 plant pathogens; Oxfordplate assay was used to measure the stability of the crude extract;HPLC was used for isolation and purification of the antifungal substances.The results showed that the fermentation broth of B10-26 had obvious antifungal activity to 13 plant pathogens,especially to Macrophomina phaseoli Ashby,with the antifungal rate of 91.67%.Although the activity of the antifungal substances declined as the temperature increased,they still produced an inhibition zone of 11 mm after 121 ℃ high temperature treatment,indicating that they had a high thermal stability.The activity did not change significantly from pH 2 to 10,indicating that they had high pH stability,and the activity showed higher under alkaline conditions.Prefractionation of the antifungal substances showed that the fermentation broth contained a variety of activity substances.