| 何紫梅 1，尹茂灵1，张 宇 1 ,2，罗学刚 1,2，王正前 3，李春华 4.噻霉酮对猕猴桃溃疡病菌的抑菌机理研究[J].广东农业科学,2023,50(3):105-112 |
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噻霉酮对猕猴桃溃疡病菌的抑菌机理研究 |
| Study on the Antimicrobial Mechanism of Benzothiazolinone Against Kiwifruit Ulcerative Pathogens |
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| DOI：10.16768/j.issn.1004-874X.2023.03.012 |
| 中文关键词: 噻霉酮 猕猴桃溃疡病 流式细胞术 生理生化 抑菌机理 防控 |
| 英文关键词: benzothiazolinone kiwifruit ulcer disease flow cytometry physiology and biochemistry antimicrobial mechanism prevention and control |
| 基金项目:四川省科技计划重点研发项目（20212YFN0035） |
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| 中文摘要: |
| 【目的】通过测定噻霉酮水乳剂处理后的丁香假单胞杆菌猕猴桃致病变种（Pseudomonas syringae pv. Actinidiae，Psa）的一系列指标变化，揭示噻霉酮对猕猴桃溃疡病菌的抑菌机理，为噻霉酮防控猕猴桃溃疡病提供理论依据。【方法】对 Psa 进行稀释 10 倍的噻霉酮处理（X10），原液噻霉酮处理（Y）以及无菌水处理（CK），利用试剂盒测定 Psa 抗氧化酶系过氧化氢酶（CAT）、超氧化物歧化酶（SOD）的酶活性，考马斯亮蓝法测定可溶性蛋白质量浓度；通过荧光显微镜和扫描电子显微镜观察 Psa 细胞膜结构的变化；通过流式细胞术进行细胞周期的检测。【结果】噻霉酮处理能明显抑制 Psa 生长和繁殖，缩短菌体细胞的对数生长期，使细胞膜破裂，细胞内容物外泄，细胞死亡量增加，最终处理组菌体量为 CK 的 5.5%；Psa 对噻霉酮存在氧化应激反应，处理后的 Psa 活性显著升高，呈先上升后下降的趋势。噻霉酮处理后 Psa 的 CAT 活性在 10 h 时到达峰值 118.795 U/mg，SOD 活性在 4 h 时达到峰值 1 060.452 U/mg。X10、Y 的菌体蛋白质量浓度在 12 h 与 CK 相比差异显著，分别降低了 47.1%、73.4%；随噻霉酮浓度的增加，G0G1 期细胞堆积越多，细胞百分比由 CK 的 28.85% 增加至Y 的 45.23%，S 期细胞比例减少，由 CK 的 58.84% 降低至 Y 的 41.86%。Y 的 G0G1 期细胞比 CK 高 56.8%，在S 期比 CK 低 23.7%。【结论】噻霉酮能使 Psa 菌体细胞膜破裂，引起 Psa 的氧化应激反应，并通过抑制菌体细胞的 DNA 合成抑制 Psa 生长，有效防控猕猴桃细菌性溃疡病。 |
| 英文摘要: |
| 【Objective】By measuring a series of index changes of Pseudomonas syringae pv. Actinidiae (Psa) after treatment with benzothiazolinone aqueous emulsion, the antibacterial mechanism of benzothiazolinone aqueous emulsion against Psa was revealed, which would provide a theoretical basis for the control of kiwifruit ulcer disease with benzothiazolinone.【Method】Psa was treated with benzothiazolinone diluted 10 times (X10), benzothiazolinone stock solution (Y) and sterile water (CK). The enzymatic activities of catalase (CAT) and superoxide dismutase (SOD) were determined
by using kits and the soluble protein content was determined by Coomassie Brilliant Blue staining. Changes in the cell membrane structure of Psa were observed by fluorescence microscopy and scanning electron microscopy and cell cycle assays were performed by flow cytometry.【Result】The study showed that Psa growth and reproduction were significantly inhibited by benzothiazolinone treatment, which shortened the logarithmic growth period of Psa, caused cell membrane rupture, cell contents leakage and increased cell death, and the final amount of mycobacteria was only 5.5% of the control. Psa had oxidative stress response to benzothiazolinone, and the enzyme activities of Psa were significantly increased after treatment, showing a trend of increasing first and then decreasing. After treatment of benzothiazolinone, CAT enzyme activity reached a peak of 118.795 U/mg at 10 h and SOD enzyme activity reached a peak of 1 060.452 U/mg at 4 h. Bacterial protein content of X10 and Y differed significantly at 12 h compared with the control, decreasing by 47.1% and 73.4%, respectively. With the increase of the concentration of benzothiazolinone, more G0G1 phase cells piled up and the percentage of cells increased from 30.27% in the control group to 45.23% in treatment group Y. The percentage of S phase cells decreased from 58.84% in the control group to 41.86% in treatment group Y. The G0G1 phase cells in treatment group Y were 56.8% higher than those in CK and 23.7% lower than those in CK in S phase.【Conclusion】Benzothiazolinone ruptures bacterial cell membranes, causes oxidative stress response in Psa, and inhibits Psa growth by inhibiting DNA synthesis in bacterial cells. It can effectively prevent and control bacterial ulcer disease of kiwifruit. |
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