College of Life Science and Agroforestry, Qiqihar University, Qiqihar, China.
Heilongjiang Provincial Technology Innovation Center of Agromicrobial Preparation Industrialization, Qiqihar, China.
Microb Biotechnol. 2024 Oct;17(10):e70026. doi: 10.1111/1751-7915.70026.
Fusarium wilt is one of the major constraints on global watermelon production, and Fusarium oxysporum f. sp. niveum (Fon) is the causative agent of Fusarium wilt in watermelon and results in severe yield and quality losses worldwide. The enhancement of antifungal activity from antagonistic bacteria against Fon is highly practical for managing Fusarium wilt in watermelon. The aim of this study was to maximize the antifungal activity of Bacillus velezensis LZN01 by optimizing fermentation conditions and analysing its regulatory mechanism via transcriptome sequencing. The culture and fermentation conditions for strain LZN01 were optimized by single-factor and response surface experiments. The optimum culture conditions for this strain were as follows: the addition of D-fructose at 35 g/L and NHCl at 5 g/L in LB medium, pH 7, and incubation at 30°C for 72 h. The fungal inhibition rate for strain LZN01 reached 71.1%. The improvement of inhibition rate for strain LZN01 in optimization fermentation was supported by transcriptomic analysis; a total of 491 genes were upregulated, while 736 genes were downregulated. Transcriptome analysis revealed that some differentially expressed genes involved in carbon and nitrogen metabolism, oxidation-reduction, fatty acid and secondary metabolism; This optimization process could potentially lead to significant alterations in the production levels and types of antimicrobial compounds by the strain. Metabolomics and UPLC/Q-Exactive Orbitrap MS analysis revealed that the production yields of antimicrobial compounds, such as surfactin, fengycin, shikimic acid, and myriocin, increased or were detected in the cell-free supernatant (CFS) after the fermentation optimization process. Our results indicate that fermentation optimization enhances the antifungal activity of the LZN01 strain by influencing the expression of genes responsible for the synthesis of antimicrobial compounds.
镰刀菌枯萎病是全球西瓜生产的主要限制因素之一,尖孢镰刀菌西瓜专化型(Fusarium oxysporum f. sp. niveum,Fon)是西瓜枯萎病的病原菌,在全球范围内导致严重的产量和品质损失。拮抗细菌对 Fon 的抑菌活性增强在防治西瓜枯萎病方面具有很高的实际应用价值。本研究旨在通过优化发酵条件和分析转录组测序来最大化解淀粉芽孢杆菌 LZN01 的抑菌活性。通过单因素和响应面实验优化了 LZN01 菌株的培养和发酵条件。该菌株的最佳培养条件为:在 LB 培养基中添加 35 g/L D-果糖和 5 g/L NH4Cl,pH 值为 7,在 30°C 下培养 72 h。菌株 LZN01 的抑菌率达到 71.1%。通过转录组分析支持了优化发酵过程中菌株 LZN01 抑菌率的提高;共有 491 个基因上调,736 个基因下调。转录组分析表明,一些差异表达基因参与了碳氮代谢、氧化还原、脂肪酸和次生代谢;这一优化过程可能会导致该菌株产生的抗菌化合物的产生水平和类型发生显著变化。代谢组学和 UPLC/Q-Exactive Orbitrap MS 分析表明,发酵优化后,抗菌化合物如表面活性剂、丰原素、莽草酸和麦角甾醇的产量增加或在无细胞上清液(CFS)中检测到。我们的结果表明,发酵优化通过影响合成抗菌化合物的基因表达来增强 LZN01 菌株的抑菌活性。