| 小麦茎基腐病生防菌P61的分离鉴定及发酵条件筛选 |
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| 引用本文：孙昊,董素芬,李雪,张佳琪,李满,李星,康占海.小麦茎基腐病生防菌P61的分离鉴定及发酵条件筛选.植物保护学报,2025,52(2):366-378 |
| DOI：10.13802/j.cnki.zwbhxb.2025.2024123 |
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| 中文摘要:为筛选对小麦茎基腐病具有拮抗作用的生防菌株，从河北省小麦根际土壤中分离拮抗细菌，以假禾谷镰孢菌Fusarium pseudograminearum为靶标菌，采用平板对峙法进行生防菌筛选，基于形态特征、生理生化特性和分子鉴定结果对筛选菌株进行鉴定，并通过单因素试验、Plackett-Burman试验及响应曲面分析法对其培养成分和发酵条件进行筛选。结果表明：从小麦根际土壤中共分离到11株菌，最终筛选出对假禾谷镰孢菌抑制效果最强的菌株P61，抑制率达到79.88%，并将其鉴定为解淀粉芽胞杆菌Bacillus amyloliquefaciens。菌株P61的发酵滤液对假禾谷镰孢菌孢子萌发的抑制率达78.86%，可分泌酯酶、纤维素酶、淀粉酶、蛋白酶及有机酸，并对多种镰孢菌表现出显著的抑制效果。对菌株P61培养成分进行筛选后确定其最佳培养基配方为甘露醇7 g/L、蛋白胨7 g/L、MgSO4 4.31 g/L；其筛选后的发酵条件为接种量3%、装液量40%、转速186 r/min、温度30.9℃、培养时间44.83 h、pH 7.0；在此发酵条件下菌株P61的生物量达6.533×109 CFU/mL，对假禾谷镰孢菌的抑制率较发酵条件筛选前提高了9.47百分点。 |
| 中文关键词:小麦茎基腐病 假禾谷镰孢菌 解淀粉芽胞杆菌 响应曲面分析法 发酵配方 发酵条件 |
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| Isolation and identification of biocontrol bacterium P61 against wheat stem rot and screening of fermentation conditions |
| Author Name | Affiliation | E-mail | | Sun Hao | College of Plant Protection, Hebei Agricultural University, Baoding 071001, Hebei Province, China | | | Dong Sufen | College of Information Science and Technology, Hebei Agricultural University, Baoding 071001, Hebei Province, China | | | Li Xue | College of Plant Protection, Hebei Agricultural University, Baoding 071001, Hebei Province, China | | | Zhang Jiaqi | College of Plant Protection, Hebei Agricultural University, Baoding 071001, Hebei Province, China | | | Li Man | College of Plant Protection, Hebei Agricultural University, Baoding 071001, Hebei Province, China | | | Li Xing | College of Plant Protection, Hebei Agricultural University, Baoding 071001, Hebei Province, China | lxkzh@163.com | | Kang Zhanhai | College of Plant Protection, Hebei Agricultural University, Baoding 071001, Hebei Province, China | xkjiating@163.com |
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| Abstract:To screen antagonistic bacterial strains against wheat crown rot, rhizosphere soil samples from Hebei Province wheat fields were collected for bacterial isolation. Fusarium pseudograminearum was selected as the target pathogen for preliminary biocontrol strain selection using dual-culture plate assays. Comprehensive characterization of the selected strain P61 was conducted through morphological observation, physiological-biochemical profiling, and molecular identification. Subsequent fermentation optimization employed single-factor experiments, Plackett-Burman design, and response surface methodology. The results demonstrated that 11 bacterial isolates were obtained, with strain P61 exhibiting the strongest antifungal activity (79.88% inhibition rate) and identified as Bacillus amyloliquefaciens. The fermentation filtrate of P61 showed 78.86% suppression rate on F. pseudograminearum conidial germination, and demonstrated enzymatic activities including esterase, cellulase, amylase, protease, and organic acid production, while displaying broad-spectrum antagonism against multiple Fusarium species. Through fermentation optimization, the optimal medium composition was determined as mannitol 7 g/L, peptone 7 g/L, and MgSO4 4.31 g/L. The ideal fermentation parameters comprised of 3% inoculation volume, 40% liquid loading volume, 186 r/min agitation, 30.9 ℃ incubation temperature, 44.83 h cultivation duration, and pH 7.0. Under fermentation conditions, P61 achieved a biomass of 6.533×109 CFU/mL with 9.47% enhancement in antifungal efficacy against F. pseudograminearum. These findings suggest that B. amyloliquefaciens P61 is a promising biocontrol agent for wheat crown rot management. |
| keywords:wheat stem basal rot Fusarium pseudograminearum Bacillus amyloliquefaciens response surface methodology fermentation formulation fermentation condition |
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