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从枯草芽孢杆菌Z-14中分离并优化对小麦全蚀病菌引起的小麦全蚀病具有活性的脂肽产量。

Isolation and yield optimization of lipopeptides from Bacillus subtilis Z-14 active against wheat take-all caused by Gaeumannomyces graminis var. tritici.

作者信息

Zhang Xuechao, Chen Xiaomeng, Qiao Xinlei, Fan Xuerui, Huo Xiaoyi, Zhang Dongdong

机构信息

College of Life Science, Hebei Agricultural University, Baoding, P. R. China.

出版信息

J Sep Sci. 2021 Feb;44(4):931-940. doi: 10.1002/jssc.201901274. Epub 2021 Jan 18.

Abstract

Wheat take-all, caused by the soil-borne fungus Gaeumannomyces graminis var. tritici, is one of the major constraints on wheat production worldwide. Bacillus subtilis Z-14 exerts significant biocontrol activity against wheat take-all, and lipopeptide antibiotics are the main antifungal substances. Herein, lipopeptide antibiotics C14-C15 iturin A, C14-C16 fengycin A, and C15-C17 fengycin B from B. subtilis Z-14 culture filtrates were separated and identified by high-performance liquid chromatography, matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry, and mass spectrometry/mass spectrometry, respectively. The optimal medium components for Z-14 lipopeptide antibiotic production were 3.85 g/L corn flour, 1.57 g/L soybean meal, 0.03 g/L FeSO ·7H O, 0.2 g/L NaH PO ·2H O, and 0.4 g/L Na HPO ·2H O. Quantification analysis by high-performance liquid chromatography showed that fengycins played a main role in antifungal activity against Gaeumannomyces graminis var. tritici. Quantitative reverse transcription polymerase chain reaction showed that lipopeptide synthesis genes fenD and ituC reached maximum expression levels after 48 h of fermentation. The strongest control of wheat take-all by Z-14 was achieved by adding 30 mL of culture filtrate per 350 g of soil in pot experiments, during which disease reduction reached 88.15%. This study provides theoretical support and a material basis for the prevention and treatment of wheat take-all disease.

摘要

由土壤传播的真菌禾顶囊壳小麦变种引起的小麦全蚀病是全球小麦生产的主要限制因素之一。枯草芽孢杆菌Z-14对小麦全蚀病具有显著的生物防治活性,脂肽抗生素是主要的抗真菌物质。在此,分别通过高效液相色谱、基质辅助激光解吸/电离飞行时间质谱和质谱/质谱,对枯草芽孢杆菌Z-14培养滤液中的脂肽抗生素C14-C15伊枯草菌素A、C14-C16丰原素A和C15-C17丰原素B进行了分离和鉴定。Z-14产脂肽抗生素的最佳培养基成分是3.85 g/L玉米粉、1.57 g/L豆粕、0.03 g/L FeSO·7H₂O、0.2 g/L NaH₂PO₄·2H₂O和0.4 g/L Na₂HPO₄·2H₂O。高效液相色谱定量分析表明,丰原素在对禾顶囊壳小麦变种的抗真菌活性中起主要作用。定量逆转录聚合酶链反应表明,脂肽合成基因fenD和ituC在发酵48小时后达到最大表达水平。在盆栽试验中,每350 g土壤添加30 mL培养滤液对小麦全蚀病的防治效果最强,病害减轻率达88.15%。本研究为小麦全蚀病的防治提供了理论支持和物质基础。

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