防御假单胞菌FD6对番茄灰霉病菌的抑菌活性及其培养条件优化

doi:10.16420/j.issn.0513-353x.2024-0829

摘要/Abstract

摘要： 防御假单胞菌FD6通过产生抗生素藤黄绿脓菌素（PLT）和2,4-二乙酰基间苯三酚（2,4-DAPG）可有效防治番茄灰霉病。细菌次生代谢物的产生水平不仅取决于菌株本身，还与其发酵条件密切相关。为提高FD6抑菌活性，从6种不同培养基中筛选出抑菌活性最强的培养基LB，以其作为基础发酵培养基采用单因素试验、响应曲面法筛选优化FD6的培养基成分及发酵条件。结果表明，最佳发酵培养基组分配比为柠檬酸钠10.81 g · L^-1、蛋白胨10.53 g · L^-1、缬氨酸5.28 g · L^-1，最佳发酵条件为pH 7.2、温度28 ℃、振荡培养120 h。在此优化条件下，FD6无菌滤液对番茄灰霉病菌的平均抑菌率达到80.26%，较优化前提高了39.39%。抗生素定量结果表明，优化后PLT产量达14.85 μg · mL^-1，较优化前提高了4.71倍。研究结果为防御假单胞菌FD6菌剂的开发及其在番茄灰霉病生物防治中的应用提供参考。

关键词: 番茄, 防御假单胞菌, 培养条件, 优化, 单因素试验, 响应曲面, 灰霉病菌

Abstract:

Pseudomonas protegens FD6 can effectively control tomato gray mold by producing antibiotics pyoluteorin and 2,4-diacetylphloroglucinol. The yield of bacterial secondary metabolites depends not only on the strain，but also the fermentation conditions. In order to improve the antifungal activity of FD6，the antagonistic ability of culture filtrates from six different media and LB filtrate displayed the best antifungal activity were assayed. Then LB were used as the basic fermentation medium to optimize the medium composition and fermentation conditions of FD6 through single-factor test and response surface method. The results showed that the optimal fermentation conditions of strain FD6 were as follows：sodium citrate 10.81 g · L^-1，peptone 10.53 g · L^-1，valine 5.28 g · L^-1，pH 7.2，28 ℃，cultivation time 120 h. After optimization，the average inhibition rate of the sterile filtrate of FD6 against Botrytis cinerea was 80.26%，representing a 39.39% increase compared to pre-optimization levels. The quantitative analysis of antibiotics revealed that the production of PLT reached 14.85 μg · mL^-1 after optimization，and caused a 4.71-fold increase compared with pre-optimization levels. Our results provide a valuable reference for the development of P. protegens FD6 bacterial agent and its application in biological control of tomato gray mold.

Key words: tomato, Pseudomonas protegens, culture condition, optimization, single factor experiment, response surface, Botrytis cinerea

宋琦, 宋晓雅, 焦永鑫, 毛子玥, 吴小刚, 张清霞. 防御假单胞菌FD6对番茄灰霉病菌的抑菌活性及其培养条件优化[J]. 园艺学报, 2025, 52(9): 2545-2553.

SONG Qi, SONG Xiaoya, JIAO Yongxin, MAO Ziyue, WU Xiaogang, ZHANG Qingxia. Antifungal Activity of Pseudomonas protegens FD6 Against Botrytis cinerea and Optimization of Its Culture Conditions[J]. Acta Horticulturae Sinica, 2025, 52(9): 2545-2553.

图/表 5

图1 6种培养基对菌株FD6发酵上清液抑制灰霉菌效果的影响不同小写字母表示差异显著（P < 0.05），下同

Fig. 1 The inhibitory effect of the fermentation supernatant of Pseudomonas protegens FD6 on Botrytis cinerea in six media Different lowercase letters indicate significant differences（P < 0.05），the same below

图2 不同碳源（A）、氮源（B）、氨基酸（C）对菌株FD6抑制灰霉菌效果的影响

Fig. 2 Effects of different carbon source（A），nitrogen sources（B）and amino acid（C）on the inhibition of Botrytis cinerea by Pseudomonas protegens FD6

图3 不同浓度无机盐对菌株FD6抑制灰霉菌效果的影响

Fig. 3 Effects of different inorganic ions on the inhibition of Botrytis cinerea by Pseudomonas protegens FD6

图4 不同培养条件对菌株FD6抑制灰霉菌的效果的影响

Fig. 4 Effects of different culture conditions on the inhibition of Botrytis cinerea by Pseudomonas protegens FD6

图5 柠檬酸钠和蛋白胨、柠檬酸钠和缬氨酸、蛋白胨和缬氨酸对抑制率交互影响的响应面图

Fig. 5 Response surface diagram of the interaction effect of sodium citrate and peptone，sodium citrate and valine，peptone and valine on the inhibition rate

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