木醋液对几种植物病原真菌的抑菌活性与作用机制

doi:10.3969/j.issn.1004-1524.20221348

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (9): 2149-2159.DOI: 10.3969/j.issn.1004-1524.20221348

木醋液对几种植物病原真菌的抑菌活性与作用机制

李彦湘¹(), 丁德东¹, 何静¹^,^*(), 张金花², 赵吉桃¹, 赵倩¹, 候彩霞¹, 朱珠¹

1.甘肃农业大学林学院,甘肃兰州 730070
2.陇南市科学技术情报研究所,甘肃陇南 742500

收稿日期:2022-09-16 出版日期:2023-09-25 发布日期:2023-10-09
作者简介:李彦湘(1997—),女,甘肃定西人,硕士研究生,主要从事经济林有害生物防治研究。E-mail:1837161121@qq.com
通讯作者: 何静,E-mail:hejing268@aliyun.com
基金资助:
甘肃省自然科学基金重点项目(20JR10RA508);甘肃农业大学科技创新基金项目(GAU-XKJS-2018-100)

Antifungal activity and mechanism of wood vinegar against several plant pathogenic fungi

LI Yanxiang¹(), DING Dedong¹, HE Jing¹^,^*(), ZHANG Jinhua², ZHAO Jitao¹, ZHAO Qian¹, HOU Caixia¹, ZHU Zhu¹

1. College of Forestry, Gansu Agricultural University, Lanzhou 730070, China
2. Longnan Science and Technology Information Research Institute, Longnan 742500, Gansu, China

Received:2022-09-16 Online:2023-09-25 Published:2023-10-09

摘要/Abstract

摘要：

为探究木醋液在植物病害生物防治中的实际应用及其对病原菌的作用机制,试验通过测定菌落生长直径和半数有效浓度(EC₅₀值)评价木醋液对几种供试植物病原真菌的抑菌活性。考察木醋液对供试敏感菌株孢子萌发、菌丝干重、细胞膜通透性、细胞壁完整性、可溶性蛋白含量、可溶性糖含量、还原性糖含量和腺嘌呤核苷三磷酸(ATP)含量等指标的影响,结合电镜观察菌体超微结构变化揭示其作用机制。结果表明:木醋液对7种供试植物病原真菌均具一定的抑制效果,对黄色镰刀菌(Fusarium culmorum)的抑菌作用表现最为显著,其EC₅₀值为4.98 μL·mL^-1,在5.5 μL·mL^-1时,木醋液对其孢子萌发和生物量抑制率分别达90.09%和95.22%。经木醋液处理后,F. culmorum的细胞结构遭到破坏,其菌丝细胞壁变薄且不完整,细胞器结构紊乱,细胞膜通透性增加;菌丝体碱性磷酸酶活性、可溶性糖含量、还原性糖含量、可溶性蛋白含量和ATP含量显著降低;碘化丙啶染色孢子后,孢子荧光强度显著增加。以上结果表明,木醋液主要通过破坏真菌细胞结构、扰乱菌体物质代谢和能量代谢来实现其抑菌效果。

关键词: 木醋液, 真菌, 黄色镰刀菌, 抑菌活性

Abstract:

To explore the practical application of wood vinegar in the biological control of plant diseases and its mechanism of action against pathogenic fungi, experiments were conducted to evaluate the antimicrobial activity of wood vinegar against several tested plant pathogenic fungi by measuring the colony growth diameter and half effective concentration (EC₅₀). The impact of wood vinegar on spore germination, mycelial dry weight, cell membrane permeability, cell wall integrity, soluble protein content, soluble sugar content, reducing sugar content, and adenosine triphosphate (ATP) content of sensitive fungal strains was assessed, and the ultrastructural changes of the fungal bodies were observed through electron microscopy to reveal its mechanism of action. The results showed that wood vinegar had certain inhibitory effects on all 7 tested plant pathogenic fungi, with the most significant inhibitory effect observed against Fusarium culmorum, with an EC₅₀ value of 4.98 μL·mL^-1, and the inhibition of spore germination and biomass at a volume fraction of 5.5 μL·mL^-1 reached 90.09% and 95.22%, respectively. After treatment with wood vinegar, the cell structure of F. culmorum was damaged, the cell walls of the mycelium became thinner and incomplete, organelle structure was disrupted, and cell membrane permeability increased. Alkaline phosphatase activity, soluble sugar content, reducing sugar content, soluble protein content and ATP content of the mycelium were significantly reduced. Additionally, the fluorescence intensity of iodine-propidium iodide stained spores increased significantly. Wood vinegar mainly achieves its antimicrobial effect by damaging the fungal cell structure and interfering with fungal substance metabolism and energy metabolism.

Key words: wood vinegar, fungus, Fusarium culmorum, antifungal activity

中图分类号:

S482.2

李彦湘, 丁德东, 何静, 张金花, 赵吉桃, 赵倩, 候彩霞, 朱珠. 木醋液对几种植物病原真菌的抑菌活性与作用机制[J]. 浙江农业学报, 2023, 35(9): 2149-2159.

LI Yanxiang, DING Dedong, HE Jing, ZHANG Jinhua, ZHAO Jitao, ZHAO Qian, HOU Caixia, ZHU Zhu. Antifungal activity and mechanism of wood vinegar against several plant pathogenic fungi[J]. Acta Agriculturae Zhejiangensis, 2023, 35(9): 2149-2159.

图/表 7

表1 木醋液对7种植物病原真菌的抑制作用

Table 1 Inhibitory effects of wood vinegar on 7 plant pathogenic fungi

病原菌 Pathogenic fungus	回归方程 Regression equation	EC₅₀/(μL·mL^-1)	相关系数 Correlation coefficient
黄色镰刀菌F. culmorum	y=2.14x+3.51	4.98	0.958 5
三线镰刀菌F. tricinctum	y=1.33x+4.06	5.10	0.948 4
杨生盾壳霉C. populicola	y=2.45x+3.22	5.33	0.989 1
木贼镰刀菌F. equiseti	y=1.77x+3.25	9.69	0.945 2
链格孢菌A. alternata	y=2.18x+2.75	10.80	0.967 0
尖孢镰刀菌F. oxysporum	y=1.41x+3.00	26.50	0.944 3
腐皮镰刀菌F. solani	y=1.77x+2.26	22.04	0.993 7

图1 不同体积分数木醋液对黄色镰刀菌孢子萌发和芽管伸长的影响柱上无相同小写字母表示差异显著(P<0.05)。下同。

Fig.1 Effects of different volume fractions of wood vinegar on spore germination and germ tube elongation of F. culmorum Data marked without the same lowercase letter indicated significant differences at P<0.05. The same as below.

图2 木醋液对黄色镰刀菌生物量的影响

Fig.2 Effect of wood vinegar on biomass of F. culmorum

图3 木醋液对黄色镰刀菌超微结构的影响 A,对照组;B,4.98 μL·mL-1 木醋液处理;C,5.5 μL·mL-1木醋液处理;CW,细胞壁;ER,内质网;V,液泡。

Fig.3 Effects of wood vinegar on ultrastructure of F. culmorum(2 000×) A, Control; B, Treatment with 4.98 μL·mL-1 wood vinegar; C, Treatment with 5.5 μL·mL-1 wood vinegar; CW, Cell wall; ER, Endoplasmic reticulum; V, Vacuole.

图4 木醋液对黄色镰刀菌细胞膜完整性的影响

Fig.4 Effects of wood vinegar on cell membrane integrity of F. culmorum

图5 木醋液对黄色镰刀菌相对渗透率和AKP活性的影响 *表示差异显著(P<0.05),**表示差异极显著(P<0.01)。下同。

Fig.5 Effects of wood vinegar on relative permeability and AKP activity of F. culmorum * indicates significant difference at P<0.05, and ** indicates significant difference at P<0.01. The same as below.

图6 木醋液对黄色镰刀菌可溶性糖、可溶性蛋白、还原性糖和ATP含量的影响

Fig.6 Effects of wood vinegar on contents of soluble sugar, soluble protein, reducing sugar and ATP of F. culmorum

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木醋液对几种植物病原真菌的抑菌活性与作用机制

Antifungal activity and mechanism of wood vinegar against several plant pathogenic fungi

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