榆黄蘑木霉病病原菌分离鉴定及植物源抑菌剂筛选

Isolation and identification of pathogen of Trichoderma disease in Pleurotus citrinipileatus and screening of plant bacteriostatic agents

    摘要
  • 摘要: 【目的】明确榆黄蘑木霉病的病原菌种类，筛选有效的植物源抑菌剂，为榆黄蘑木霉病的科学诊断和田间防治提供理论依据。【方法】从重庆市万州区龙驹镇灯台村智富森华农业专业合作社大棚食用菌栽培基地采集榆黄蘑染病菌包，采用组织分离法对榆黄蘑染病菌包中的病原菌进行分离，通过形态学特征和分子生物学方法对病原菌进行种类鉴定。选取32种常见药用植物通过醇溶液提取获得提取液；采用菌丝生长速率法测定不同药用植物提取液对病原菌菌丝生长的影响；通过最低抑菌浓度（MIC）和最低杀菌浓度（MBC）确定药用植物提取液的最佳使用浓度；通过孢子萌发试验评估药用植物提取液对病原菌的抑制效果。【结果】从榆黄蘑染病菌包中共分离出2株木霉菌，通过形态学特征和分子生物学方法鉴定为长枝木霉（Trichoderma longibrachiatum）和深绿木霉（Trichoderma atroviride）。抑菌试验发现，黄连和甘草提取液对长枝木霉的抑制效果较佳，平均菌丝生长抑制率分别为94.54%和81.90%,MIC均为15.60 mg/mL,MBC均为31.25 mg/mL，孢子萌发抑制率最高分别达97.73%和96.95%，抑制中浓度(EC50)分别为1.0670和2.2260 mg/mL；黄连提取液对深绿木霉的综合效果最佳，平均菌丝生长抑制率为65.82%,MIC为250.00 mg/mL，孢子萌发抑制率达89.21%,EC50为34.1080 mg/mL。【结论】长枝木霉和深绿木霉是污染榆黄蘑的主要病原菌。黄连和甘草提取液对2种木霉菌均有较好的抑制效果，可作为植物源抑菌剂在食用菌种植中推广应用。

     

    Abstract: 【Objective】To clarify the pathogen species of Trichoderma disease in Pleurotus citrinipileatus and screen effective plant bacteriostatic agents,which could provide theoretical basis for scientific diagnosis and field control of Trichoderma disease in Pleurotus citrinipileatus. 【Method】The contaminated packets of Pleurotus citrinopileatus were collected from the edible fungus cultivation base of Zhifusenhua Agricultural Cooperative in Dengtai Village,Longju Town,Wanzhou District,Chongqing City. The pathogenic bacteria in the contaminated bags of Pleurotus citrinopileatus were isolated by tissue separation method,and the pathogens were identified by morphological characteristics and molecular biology methods. A total of 32 kinds of common medicinal plants were extracted by alcohol solution to form extract. The effects of different medicinal plant extracts on the mycelium growth of pathogens were determined by mycelium growth rate method. The optimal concentration of fungicides was determined by minimum inhibitory concentration（MIC） and minimal bactericidal concentration（MBC）. The inhibitory effects of fungicides on pathogens were evaluated by spore germination experiment. 【Result】Two Trichoderma strains were isolated from the contaminated packets of Pleurotus citrinopileatus and identified as Trichoderma longibrachiatum and Trichoderma atroviride. The results of bacteriostatic experiment showed that the extracts of Coptis chinensis Franch. and Glycyrrhiza uralensis Fisch. had the better inhibitory effects on Trichoderma longibrachiatum,the average mycelium growth inhibition rates were 94.54% and 81.90%,both MIC and MBC were 15.60 and 31.25 mg/mL respectively,and the highest spore germination inhibition rates were 97.73% and 96.95% respectively. The median inhibitory concentration(EC50) were 1.0670 and 2.2260 mg/mL respectively. The comprehensive effects of Coptis chinensis Franch. extract on Trichoderma atroviride was the best,with an average mycelium growth inhibition rate of 65.82%,MIC of 250.00 mg/mL,spore germination inhibition rate of 89.21% and EC50 of 34.1080 mg/mL. 【Conclusion】T. longibrachiatum and Trichoderma atroviride are the main pathogens that contaminate P. citrinopileatus. The extracts of Coptis chinensis Franch. and Glycyrrhiza uralensis Fisch. can effectively inhibit these 2 pathogens and can be used as plant bacteriostatic agents in the cultivation of edible fungi.

     

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