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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2017, Vol. 37 Issue (5): 76-86    DOI: 10.13523/j.cb.20170510
    
Screening and Identification of Biocontrol Fungi against Didymella bryoniae and Optimization of Fermentation Conditions
ZHANG Xu-hui1, ZHANG Hong-nan2, LI Yong1, WANG Wen-qiang1
1. College of Resources and Environment, Southwest University, Chongqing 400715, China;
2. College of Horticulture and Landscape Architecture, Southwest University, Chongqing 400715, China
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Abstract  

In order to screen the biocontrol strain with antagonistic effects on Didymella bryoniae and enhance the inhibitory efficacy of the fermentation broth, antagonistic fungi was screened using the dual culture assay and shaking flask culture experiment. Subsequently, biocontrol strain was identified according to morphological character and 18S rDNA sequence analysis, and observation of parasitic effect of antagonistic strain on Didymella bryoniae was made by scanning electron microscope. The fermentation conditions were optimized according to single factor test and response surface methodology. In addition, the biocontrol efficacy of Bjerkandera adusta against Didymella bryoniae in plants was determined in greenhouse. The results reveal that the strain M1 has the highest inhibitory rates to Didymella bryoniae, reaching up to 59.56% and 49.56% in the dual culture assay and in shaking flask culture experiment separately. The morphological characteristics of M1 are consistent with those of Bjerkandera adusta and 18S rDNA sequence phylogenetic analysis reveal that M1 appears a sister lineage to Bjerkandera adusta and gathers together in a phylogenetic tree. So the strain M1 is identified as Bjerkandera adusta. Additionally, M1 is able to penetrate Didymella bryoniae mycelium directly. After optimized, the fermentation conditions are C/N of 7.1, pH of 7.4, the bottling quantity of 44%, fermentation time of 19d, the rotate speed of 180r/min, culture temperature of 29℃with the inhibitory rate of the fermentation broth increased from 49.56% to 52.64%. The biocontrol efficiency of Bjerkandera adusta against Didymella bryoniae is up to 74.3% in greenhouse, which is higher than that of treatment with carbendazim. These results demonstrate that as a biocontrol fungus, Bjerkandera adusta has an antagonistic effect on Didymella bryoniae, and also has potential value to develop into antiseptic agent in the field application.

Key words： Bjerkandera adusta      Didymella bryoniae      Antagonism      Condition optimization      Inhibitory rate     
Received: 24 November 2016      Published: 25 May 2017
ZTFLH:  Q815  
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ZHANG Xu-hui, ZHANG Hong-nan, LI Yong, WANG Wen-qiang. Screening and Identification of Biocontrol Fungi against Didymella bryoniae and Optimization of Fermentation Conditions. China Biotechnology, 2017, 37(5): 76-86.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20170510     OR     https://manu60.magtech.com.cn/biotech/Y2017/V37/I5/76

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