姜精油和柠檬醛对生姜枯萎病菌的抑菌作用及盆栽防效

周丽荣; 熊诗洁; 张玲玲; 马慧慧; 朱学栋; 尹军良; 刘奕清

doi:10.12357/cjea.20230230

姜精油和柠檬醛对生姜枯萎病菌的抑菌作用及盆栽防效

doi: 10.12357/cjea.20230230

1.
长江大学园艺园林学院/香辛作物研究院　荆州　434025
2.
重庆市渝东南农业科学院　渝东　408000
3.
长江大学农学院　荆州　434025

基金项目: 重庆英才优秀科学项目(2022CQYC027)、调味品产业技术体系生姜品种选育与丰产技术研究推广(2022-2023-07)和湖北省重点研发计划项目(2022BBA0061, 2021BBA096)资助

详细信息

作者简介:
周丽荣, 主要研究方向为植物病害防控。E-mail:1832721032@qq.com

通讯作者:
刘奕清, 主要研究方向为生姜生物育种与病害防控。E-mail: liung906@163.com

中图分类号: S435.72
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出版历程
- 收稿日期: 2023-04-28
- 录用日期: 2023-08-21
- 修回日期: 2023-09-19
- 网络出版日期: 2023-10-09

Control effect and mechanism of ginger essential oil and citral on ginger Fusamum wilt

1.
College of Horticulture and Gardening / Spice Crops Research Institute, Yangtze University, Jingzhou 434025, China
2.
Chongqing Southeast Academy of Agricultural Sciences, Chongqing 408000, China
3.
College of Agriculture, Yangtze University, Jingzhou 434025, China

Funds: This study was supported by the Chongqing Yingcai Excellent Science Project (2022CQYC027), Research and Promotion of Ginger Variety Selection and Productivity Technology (2022-2023-07) and Hubei Provincial Key R&D Projects (2022BBA0061, 2021BBA096).

More Information

Corresponding author: E-mail: liung906@163.com

摘要

摘要: 生姜枯萎病是由尖孢镰刀菌(Fusarium oxysporum)引起的的土传病害, 防控极为困难。为了探究环境友好和安全的植物源生物农药, 本研究利用平板抑菌试验和孢子萌发试验研究了姜精油和柠檬醛对生姜枯萎病菌尖孢镰刀菌的生长抑制作用, 并通过盆栽试验进一步验证了姜精油和柠檬醛对生姜枯萎病菌尖孢镰刀菌的防控效果。结果显示, 姜精油和柠檬醛处理可显著抑制生姜枯萎病菌尖孢镰刀菌的菌丝生长与孢子萌发, 2 g·L⁻¹姜精油和0.5 g·L⁻¹柠檬醛对生姜枯萎病菌尖孢镰刀菌菌丝生长的抑制率分别为82.0%与100%, 孢子萌发抑制率分别为34.7%与95.0%; 扫描电镜结果表明, 姜精油和柠檬醛处理生姜枯萎病菌尖孢镰刀菌菌丝体3 d后, 其菌丝体表现出不同程度的弯曲、褶皱和凹陷; PI染色结果发现, 姜精油和柠檬醛处理严重破坏了生姜枯萎病菌尖孢镰刀菌细胞膜的完整性和通透性, 从而导致胞浆流失、胞外电导率、蛋白质、核酸与丙二醛含量急剧增加, 麦角固醇含量减少, 进而减弱了生姜枯萎病菌尖孢镰刀菌的致病力。接种生姜枯萎病菌尖孢镰刀菌15 d后, 2 g·L⁻¹姜精油和0.5 g·L⁻¹柠檬醛处理对生姜枯萎病的防控效果分别为32.7%、42.3%, 0.5 g·L⁻¹柠檬醛处理与8 g·L⁻¹百菌清防控效果(47.1%)无显著差异。综上, 姜精油和柠檬醛对生姜枯萎病菌尖孢镰刀菌的生长有显著抑制作用, 并对生姜枯萎病有较好的防治效果, 研究结果可为开发新型植物源抑菌剂防控生姜枯萎病提供理论依据。
- 生姜油 /
- 柠檬醛 /
- 枯萎病 /
- 生姜 /
- 抑菌活性
Abstract: Fusarium wilt in ginger plants is primarily caused by Fusarium oxysporum, and is extremely difficult to control. Yet, chemical agents are an effective means to manage Fusarium wilt. To investigate environmentally friendly and safe plant-derived biopesticides, this study evaluated the inhibitory effects of ginger essential oil (GEO) and citral on F. oxysporum FOX-1 using the mycelium growth rate method and the spore germination method. First, the mycelial growth rate method was used to determine the inhibitory effect of various concentrations of GEO and citral on F. oxysporum FOX-1 mycelial growth. The lowest concentrations of GEO and citral that effectively prevented mycelial growth on a potato dextrose agar (PDA) plate after 48 h was recorded as the minimum inhibitory concentration (MIC). Following which, the spore germination method was used to assess the effects of GEO and citral at different concentrations (0, 1/2 MIC, MIC) on the spore number and spore germination of F. oxysporum FOX-1, respectively. The scanning electron microscope (SEM) was used to observe the mycelial morphology of F. oxysporum FOX-1, and propidium iodide (PI) staining was used to assess damage to its cell membrane. Furthermore, the effects of GEO and citral on the cell integrity and permeability of F. oxysporum FOX-1 were evaluated through experiments that measured the changes of the relative electrical conductivity, proteins, nucleic acids, malondialdehyde, and ergosterol. The effects of MIC GEO and citral on controlling Fusarium wilt in ginger in a pot experiment were determined 15 d after inoculation with F. oxysporum FOX-1. The results indicated that 2 g∙L⁻¹ GEO and 0.5 g∙L⁻¹ citral PDA plates significantly inhibited the mycelial growth of F. oxysporum FOX-1, with EC₅₀ values of 1.102 g∙L⁻¹ and 0.141 g∙L⁻¹ for GEO and citral, respectively. This indicates that both GEO and citral exhibited dose-dependent effects, with MIC values of 2 g∙L⁻¹ and 0.5 g∙L⁻¹, respectively. In addition, these treatments significantly inhibited the germination of F. oxysporum FOX-1 spores in the treatment of Fusarium wilt disease of ginger compared to the control (CK), particularly the 1/2 MIC versus MIC treatment. Compared to the CK, the 1/2 MIC and MIC GEO treatment reduced the number of F. oxysporum FOX-1 spores by 35.6% and 59.3%, respectively. Similarly, the 1/2 MIC and MIC citral treatment reduced the number of F. oxysporum FOX-1 spores by 61.0% and 78.0%, respectively. After 12 h of treatment, the germination rate of F. oxysporum FOX-1 spores in the 1/2 MIC and MIC GEO treatments reduced by 20.4% and 34.7%; whereas the germination rates in the 1/2 MIC and MIC citral treatments reduced by 86.1% and 95.0%, respectively. After 3 d of GEO and citral treatment, the SEM results showed that the cell wall and cell membrane of F. oxysporum FOX-1 were damaged, and could not maintain the normal linear morphology of mycelium, and showed different degrees of curvature, folds and depressions. Furthermore, the PI staining revealed that the GEO and citral treatment severely damaged the integrity and permeability of the cell membrane of F. oxysporum FOX-1, resulting in a significant increase in the number of spores. Moreover, this treatment resulted in a sharp increase in cytoplasmic loss, extracellular conductivity, protein, nucleic acid and malondialdehyde content in the damaged F. oxysporum FOX-1 cells. After 3 d of treating the mycelium, high concentrations of GEO (2 g∙L⁻¹) and eugenol (2 g∙L⁻¹) reduced the ergosterol content of F. oxysporum FOX-1 by 27.0% and 45.2%, respectively, compared with that of CK. Notably, GEO and citral treatments also weakened the pathogenicity of F. oxysporum FOX-1. Moreover, after 15 d of inoculation with F. oxysporum FOX-1, the 2 g∙L⁻¹ GEO and 0.5 g∙L⁻¹ citral treatments exhibited efficacy rates of 32.7% and 42.3%, respectively, which 0.5 g∙L⁻¹ citral treatments was not significantly different from that of the positive control, chlorothalonil, which exhibited an efficacy rate of 47.1%. In summary, GEO and citral had significant inhibitory effect on the growth of F. oxysporum FOX-1 and could control Fusamum wilt in gingers. These findings could lay the foundation for developing botanical antifungal agents for management of Fusamum wilt.
- Ginger essential oil /
- Citral /
- Fusarium wilt /
- Zingiber offificinale /
- Antibacterial activity

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图 1 不同培养时间与不同质量浓度姜精油(A)和柠檬醛(B)对生姜枯萎病菌尖孢镰刀菌菌丝生长的抑菌作用

CK: 空白对照; 30%E: 阴性对照, 30% (质量体积分数)乙醇; Chtl.: 阳性化学药物对照, 百菌清(8 g∙L⁻¹). CK: blank control; 30%E: negative control, 30% (v/v) ethanol; Chtl.: positive chemical control, chlorothalonil (8 g∙L⁻¹).

Figure 1. Effect of different concentrations of ginger essential oil (A) and citral (B) on mycelial growth of F. oxysporum FOX-1 in different incubation times

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图 2 不同质量浓度姜精油和柠檬醛对生姜枯萎病菌尖孢镰刀菌菌丝形态的影响

A: 空白对照组; B: 1/2 MIC姜精油处理; C: 1/2 MIC柠檬醛处理; D: MIC姜精油处理; E: MIC柠檬醛处理。A: blank control group; B: 1/2 MIC GEO treatment; C: 1/2 citral treatment; D: MIC GEO treatment; E: MIC citral treatment. SEM HV: 20.0 kv; SEM MAG : 5.00 kx.

Figure 2. Effect of different concentrations of ginger essential oil and citral on mycelial morphology of F. oxysporum FOX-1

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图 3 荧光显微镜观察生姜枯萎病菌尖孢镰刀菌表型

A: 空白对照组; B: 1/2 MIC姜精油处理; C: 1/2 MIC柠檬醛处理; D: MIC姜精油处理; E: MIC柠檬醛处理; 显微镜放大倍数10×40。A: blank control group; B: 1/2 MIC GEO treatment; C: 1/2 citral treatment; D: MIC GEO treatment; E: MIC citral treatment; 10×40 magnification of microscope was 10×40 times.

Figure 3. Phenotype of F. oxysporum FOX-1 observed by fluorescence microscopy

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图 4 不同质量浓度姜精油和柠檬醛处理后生姜枯萎病菌尖孢镰刀菌细胞完整性的变化

A: 姜精油处理组电导率; B: 柠檬醛处理组电导率; C: 姜精油处理组蛋白质含量; D: 柠檬醛处理组蛋白质含量; E: 姜精油处理组核酸含量; F: 柠檬醛处理组核酸含量。A: Conductivity of ginger essential oil treated group; B: Conductivity of citral treated group; C: Protein content of ginger essential oil treated group; D: Protein content of citral treated group; E: Nucleic acid content of ginger essential oil treated group; F: Nucleic acid content of citral treated group.

Figure 4. Changes in cellular integrity of F. oxysporum FOX-1, after treatment with different concentrations of ginger essential oil and citral

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图 5 不同质量浓度姜精油和柠檬醛处理对生姜枯萎病菌尖孢镰刀菌MDA与麦角固醇含量的影响

A: 姜精油处理组MDA含量; B: 柠檬醛处理组MDA含量; C: 姜精油处理组麦角固醇含量; D: 柠檬醛处理组麦角固醇含量。A: MDA content in ginger essential oil treated group; B: MDA content in citral treated group; C: Ergosterol content in ginger essential oil treated group; D: Ergosterol content in citral treated group.

Figure 5. Effect of different concentrations of ginger essential oil and citral on MDA and ergosterol content of F. oxysporum FOX-1

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表 1 姜精油和柠檬醛对生姜枯萎病菌尖孢镰刀菌的抑菌效果

Table 1. Inhibitory effects of ginger essential oil and citral on F. oxysporum FOX-1

处理 Treatment	浓度 Concentration (g·L⁻¹)	菌落直径 Colonydia meter (cm)	抑菌率 Inhibition rate (%)	毒力方程 Virulence equation	半最大效应浓度 Median effective concentration EC₅₀ (μL·L⁻¹)	R²	最小抑菌量 minimal inhibitory concentration MIC (g·L⁻¹)
姜精油 Ginger essential oil	0	4.7±0.1 a	0.0±0.0 f	y=25.929x+9.395	1.102	0.876	2
	30% ethanol	4.6±0.4a	0.0±0.0f
	0.125	4.4±0.1 ab	7.1±1.1 e
	0.25	3.9±0.1 c	18.2±1.3 d
	0.5	3.7±0.1 cd	24.3±3.2 cd
	1	3.4±0.2 d	32.1±5.3 c
	2	1.4±0.1 e	82.0±6.1 b
	4	0.6±0.0 f	100.0±0.0 a
百菌清 Chlorothalonil (Chtl.)	8	0.6±0.0 f	100.0±0.0 a	\	\	\	\
柠檬醛 Citral	0	4.9±0.1 a	0.0±0.0 d	y=12.178x−18.943	0.141	0.899	0.5
	30% ethanol	4.7±0.3 a	0.0±0.0 d
	0.125	3.2±0.1 b	40.2±3.1 c
	0.25	0.9±0.2 c	93.2±4.1 b
	0.5	0.6±0.0 d	100.0±0.0 a
	1	0.6±0.0 d	100.0±0.0 a
同列不同字母表示差异显著(P<0.05)。Different letters in the same column meant significant difference at 0.05 level.

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表 2 不同质量浓度姜精油和柠檬醛对生姜枯萎病菌尖孢镰刀菌孢子萌发的影响

Table 2. Effect of different concentrations of ginger essential oil and citral on spore germination of F. oxysporum FOX-1

处理 Treatment	孢子数量 Number of spores (×10⁴ ml L⁻¹)	孢子萌发率 Spore germination (%)
空白对照 Blank control	5.9±0.5a	89.3±1.9a
阳性对照 Negative control	5.4±0.5a	90.7±1.8a
阳性药物对照 Positive chemical control	1.8±0.3cd	9.0±0.1e
1/2MIC 姜精油 1/2MIC ginger essential oil	3.8±0.6b	71.1±1.5b
1/2 MIC 柠檬醛 1/2 MIC citral	2.3±0.2c	12.4±1.9d
MIC 姜精油 MIC ginger essential oil	2.4±0.5c	58.3±1.7c
MIC 柠檬醛 MIC citral	1.3±0.3d	4.5±0.1f
同列不同字母表示差异显著(P<0.05)。CK: 空白对照; 30%E: 阴性对照, 30% (质量体积分数)乙醇; Chtl.: 阳性化学药物对照, 百菌清(8 g∙L⁻¹)。Different letters in the same column meant significant difference at 0.05 level. CK: blank control; 30%E: negative control, 30% (v/v) ethanol; Chtl.: positive chemical control, chlorothalonil (8 g∙L⁻¹).

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表 3 姜精油和柠檬醛处理对生姜枯萎病的盆栽防治效果

Table 3. The control effect of ginger essential oil and citral on ginger Fusarium wilt

处理 Treatment	浓度 Concentration (g·L⁻¹)	病情指数 Disease Index (%)	防控效果 Control Efficacy (%)
空白对照 Blank control group	0	5.0±0.7d	0±0.0c
阳性对照 Positive control	0	68.1±0.8a	0±0.0c
阳性药物对照 Positive chemical controls	8	36.0±1.9c	47.1±4.5a
姜精油 Ginger Essential Oil	2	45.8±1.5b	32.7±3.5b
柠檬醛 Citral	0.5	39.3±2.1c	42.3±4.2a
同列不同字母表示差异显著(P<0.05)。Different letters in the same column meant significant difference at 0.05 level.

下载: 导出CSV

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