异辛醇对番茄灰霉病菌的抑菌机理研究

doi:10.16420/j.issn.0513-353x.2024-0329

摘要/Abstract

摘要：

前期研究发现异辛醇能够显著抑制番茄灰霉病菌（灰葡萄孢Botrytis cinerea）菌丝的生长。为进一步明确异辛醇对番茄灰霉病菌的抑菌机理，采用双皿对扣法测定异辛醇对灰葡萄孢菌丝生长、孢子萌发和菌核形成的影响，利用激光扫描共聚焦显微技术观察异辛醇处理后灰葡萄孢体内DNA含量和活性氧自由基水平变化，采用紫外分光光度计检测异辛醇对番茄灰霉病菌菌丝酶活性及产草酸能力的影响。测定了异辛醇对番茄果实灰霉病的防效，并检测了异辛醇对番茄果实防御酶活性的影响。结果表明：异辛醇对番茄灰霉病菌菌丝生长有明显抑制作用。20 μL · L^-1异辛醇处理后，灰霉病菌孢子萌发抑制率达100%，菌核的形成被显著抑制；灰霉病菌菌丝DNA荧光强度值明显降低，而活性氧自由基的荧光强度值显著提高。番茄果实中的超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、多聚半乳糖醛酸酶（PG）、β-1,3-葡聚糖酶（β-1,3-GA）和几丁质酶（CHI）活性均低于对照；毒力因子草酸的分泌能力显著降低。异辛醇对番茄灰霉病的防治效果为77.19%，可以提高番茄果实体内防御酶SOD、CAT、CHI、过氧化物酶（POD）和苯丙氨酸解氨酶（PAL）活性。异辛醇通过破坏番茄灰霉病菌菌丝的正常生理活性，有效抑制番茄灰霉病菌的生长，还可以诱导番茄果实的抗病性。

关键词: 番茄, 异辛醇, 灰霉病菌, 抑菌效果, 防御酶

Abstract:

Previous findings showed that isooctyl alcohol could effectively inhibit Botrytis cinerea mycelial growth. To clarify the antifungal mechanism of isooctyl alcohol on Botrytis cinerea，the inhibition effects of isooctyl alcohol on mycelia growth，spore germination and sclerotium growth were detected by using two-sealed-base-plate method. After treatment of isooctyl alcohol，the DNA content and reactive oxygen species in B. cinerea were detected by using a laser scanning confocal microscope. The effects of isooctyl alcohol on the mycelase activity and oxalic acid production were measured using ultraviolet spectrophotometry. The control effect of isoctyl alcohol on tomato gray mold was conducted. The effects of isoctyl alcohol on the activities of defense enzymes were further detected. The results showed that isoctyl alcohol had significant inhibition effects on the mycelial growth. After treatment with isooctyl alcohol at the concentration of 20 μL · L^-1，spore germination was completely inhibited with the inhibition rate of 100%，and sclerotium was also significantly inhibited. Fluorescence intensity of DNA content in B. cinerea was dramatically reduced with the increase of reactive oxygen species level. The activities of superoxide dismutase（SOD），catalase（CAT），polygalacturonase（PG），β-1,3-glucanase（β-1,3-GA）and chitinase（CHI）were all decreased in B. cinerea. Oxalic acid（OA），an important virulence factor，was inhibited significantly in B. cinerea. In addition，the control effect of isooctyl alcohol on tomato gray mold was 77.19%. Isooctyl alcohol also enhanced SOD，CAT，CHI，peroxidase（POD）and phenylalanine aminolyase（PAL）activities in tomato. Therefore，isooctyl alcohol could effectively inhibit B. cinerea growth by interfering in the normal physiological activity，and also induce resistance to B. cinerea in tomato fruit.

Key words: tomato, isooctyl alcohol, Botrytis cinerea, antifungal effect, defense enzyme

王春伟, 王燕, 段天坤, 苏雅馨, 袁胜楠, 任璐, 赵晓军, 王美琴. 异辛醇对番茄灰霉病菌的抑菌机理研究[J]. 园艺学报, 2025, 52(3): 761-772.

WANG Chunwei, WANG Yan, DUAN Tiankun, SU Yaxin, YUAN Shengnan, REN Lu, ZHAO Xiaojun, WANG Meiqin. Antifungal Mechanism of Isooctyl Alcohol on Botrytis cinerea Causing Tomato Gray Mold[J]. Acta Horticulturae Sinica, 2025, 52(3): 761-772.

图/表 8

图1 不同浓度（20、50、100、200和500 μL · L-1）异辛醇处理对番茄灰霉病菌菌丝生长的影响 ***代表经Student’s t-test 检验各处理与对照间在0.001水平有显著差异

Fig. 1 Effects of isooctyl alcohol different concentrations（20，50，100，200 and 500 μL · L-1）on mycelial growth of Botrytis cinerea of tomato *** represented significant differences between treatment and control at 0.001 levels as calculated by Student’s t-test

图2 异辛醇（20 μL · L-1）对番茄灰霉病菌孢子萌发的影响

Fig. 2 Effect of isooctyl alcohol（20 μL · L-1）on spore germination of Botrytis cinerea of tomato

图3 异辛醇（20 μL · L-1）处理对番茄灰霉病菌菌核质量和数量的影响数据均以平均值 ± 标准误（n = 3）表示，采用Student’s t-test检验比较差异显著性，*** P < 0.001

Fig. 3 Effect of isooctyl alcohol（20 μL · L-1）on sclerotium weight and sclerotium number of Botrytis cinerea of tomato Values are mean ± standard error（n = 3）. Statistical analysis was conducted by Student’s t-test method. *** P < 0.001

图4 异辛醇（20 μL · L-1）处理对番茄灰霉病菌DNA含量和活性氧自由基水平（ROS）的影响

Fig. 4 Effect of isooctyl alcohol（20 μL · L-1）on DNA content and ROS of Botrytis cinerea

图5 异辛醇（20 μL · L-1）处理对番茄灰霉病菌菌丝酶活性的影响 *和***分别代表 Student’s t-test检验在P < 0.05和P < 0.001水平上处理与对照有显著差异

Fig. 5 Effect of isooctyl alcohol（20 μL · L-1）on enzyme activities of Botrytis cinerea * and *** represented significant differences at 0.05 and 0.001 levels as calculated by Student’s t-test

图6 异辛醇（20 μL · L-1）处理对番茄灰霉病菌草酸含量的影响数据均以平均值 ± 标准误（n = 3）表示，采用Student’s t-test检验比较差异显著性，** P < 0.01，***P < 0.001

Fig. 6 Effect of isooctyl alcohol（20 μL · L-1）on oxalic acid content of Botrytis cinerea Values are mean ± standard error（n = 3）. Statistical analysis was conducted by Student’s t-test method. ** P < 0.01，***P < 0.001

图7 异辛醇（20 μL · L-1）对番茄灰霉病发病率和病斑直径的影响数据均以平均值 ± 标准误（n = 3）表示，采用Student’s t-test检验比较差异显著性，***P < 0.001

Fig. 7 Effects of isooctyl alcohol（20 μL · L-1）on incidence and lesion diameter of tomato Values are mean ± standard error（n = 3）. Statistical analysis was conducted by Student’s t-test method. ***P < 0.001

图8 异辛醇（20 μL · L-1）处理对番茄防御酶活性的影响 *、**和***分别代表 Student’s t-test 检验在P < 0.05，P < 0.01和P < 0.001水平上有显著差异

Fig. 8 Effects of isooctyl alcohol on the activities of tomato defense enzyme *，** and *** represented significant differences at 0.05，0.01 and 0.001 levels as calculated by Student’s t-test

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