海洋黏细菌防治甘蔗梢腐病的筛选评价

Screening and evaluation of marine myxobacteria against sugarcane pokkah boeng

  • 摘要: 【目的】明确来源于广西北部湾的海洋黏细菌对甘蔗梢腐病病原真菌的生物防治潜力,为研发有效防治甘蔗梢腐病的生物农药提供科学依据和物质基础。【方法】以来源于广西北部湾不同海洋生境的12种62株黏细菌为研究对象,以2株广西蔗区甘蔗梢腐病优势病原菌甘蔗镰孢菌(Fusarium sacchari)9DF-3-2和FS-2-1为靶标,通过平板对峙培养法评价黏细菌对甘蔗镰孢菌的拮抗作用,初步筛选出潜力活性菌株;利用透析膜对峙培养法、菌丝裂解法和菌丝生长速率法等从拮抗作用方式、对病原菌菌丝生长的影响等方面评价潜力活性菌株对甘蔗镰孢菌的生物防治潜力。【结果】通过平板对峙培养法从62株黏细菌中发现47株黏细菌对2株甘蔗镰孢菌9DF-3-2和FS-2-1具有明显的抑制作用,其活性菌株率达75.8%,有18株黏细菌对2株甘蔗镰孢菌的抑菌直径大于1.000 cm,其中以叶柄黏球菌(Myxococcus stipitatus)T163和蜂窝囊菌(Melittangium sp.)T269的抑制作用最强。黏细菌胞外发酵液对甘蔗镰孢菌菌丝裂解试验结果显示,黏细菌T163和T269的胞外发酵液对2株甘蔗镰孢菌的菌丝抑制作用最明显;透析膜平板隔离试验结果显示,黏细菌T269、T163和T071受透析膜隔离后对甘蔗镰孢菌仍具有很好的抑制作用;黏细菌发酵提取物抗菌活性测定结果显示,黏细菌T163和T269的发酵提取物对2株甘蔗镰孢菌均具有较好的抑制作用,其中浓度为100.0 μg/mL的T163发酵提取物的抑菌活性最好,其对9DF-3-2和FS-2-1的抑制中浓度(EC50)分别为13.97和84.58 μg/mL。【结论】来源于广西北部湾的海洋黏细菌T163和T269对甘蔗梢腐病病原菌甘蔗镰孢菌表现出较好的生物活性,具有开发成绿色生物防治剂的潜力。

     

    Abstract: 【Objective】The purpose of the study was to clarify the biological control potential of marine myxobacteria from Beibu Gulf of Guangxi against the pathogenic fungi of sugarcane pokkah boeng,and to provide scientific bases and material foundations for the development of biopesticides for the effective control of sugarcane pokkah boeng.【Method】 A total of 62 strains of 12 species of myxobacteria from different marine habitats in Beibu Gulf of Guangxi were used as research objects,and two dominant pathogens of sugarcane pokkah boeng in Guangxi,Fusarium sacchari 9DF-3-2 and FS-2-1,were targeted. The antagonistic effect of myxobacteria on F. sacchari was evaluated by plate confrontation culture method,and the potential active strains were preliminarily screened out. The biological control potential of potential active strains against F. sacchari was evaluated by the dialysis membrane confrontation culture method,mycelia lysis method,and mycelia growth rate method from the aspects of antagonistic action mode and effect on mycelia growth of the pathogen.【Result】By plate confrontation culture method,47 strains of myxobacteria from 62 strains of myxobacteria showed obvious inhibition effect on F. sacchari 9DF-3-2 and FS-2-1,with an active strain rate of 75.8%. Eighteen strains of myxobacteria inhibited two strains of F. sacchari with an inhibitory diameter greater than 1.000 cm,among which the strongest inhibition effect was observed in Myxococcus stipitatus T163 and Melittangium sp. T269. The results of myxobacteria extracellular fermentation broth on F. sacchari mycelium lysis test showed that the extracellular fermentation broth of myxobacteria T163 and T269 had the most significant inhibition effect on the mycelium of two F. sacchari strains. The results of dialysis membrane plate isolation test showed that myxobacteria T269,T163 and T071 still had good inhibition effect on F. sacchari after being isolated by dialysis membrane. The antimicrobial activity determination result of the fermentation extracts of myxobacteria showed that the fermentation extracts of myxobacteria T163 and T269 had good inhibition effect on two strains of F. sacchari,among which the best inhibitory activity was achieved by the fermentation extract of T163 at a concentration of 100.0 μg/mL,and its inhibitory median concentration(EC50)to 9DF-3-2 and FS-2-1 were 13.97 and 84.58 μg/mL,respectively.【Conclusion】The marine myxobacteria T163 and T269 from Beibu Gulf of Guangxi show good bioactivity against F. sacchari,the pathogen of pokkah boeng,and have the potential to be developed into green biocontrol agents.

     

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