| 黄芩乙醇提取物对猕猴桃溃疡病致病菌抑菌机理研究 |
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| 引用本文： | 裴沪荣,卢明秀,龙力,龙章富.黄芩乙醇提取物对猕猴桃溃疡病致病菌抑菌机理研究[J].南方农业学报,2022,53(2):477-485. |
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| 作者姓名： | 裴沪荣 卢明秀 龙力 龙章富 |
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| 作者单位： | 四川大学生命科学学院, 成都 610064 |
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| 基金项目： | 四川大学-泸州市战略合作项目;国家重点研发计划 |
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| 摘 要： | 【目的】探究黄芩乙醇提取物（Ethanol extract of Scutellaria baicalensis，ESB）对猕猴桃溃疡病病原菌丁香假单胞菌猕猴桃致病变种（Pseudomonas syringae pv.actinidiae，Psa）的抑菌机理，为开发新型植物源杀菌剂提供理论依据。【方法】以Psa为研究对象，采用琼脂打孔法评估Psa对ESB的敏感性；通过梯度稀释法确定液体培养时ESB对Psa的最低抑菌浓度（MIC）和最低杀菌浓度（MBC）。探究1/4MIC、1/2MIC、3/4MIC和MIC下ESB对Psa的作用效果。通过菌液光密度绘制Psa生长曲线、电导率仪测定培养液上清电导率、考马斯亮蓝法检测胞外可溶性蛋白含量、碱性磷酸酶（AKP）试剂盒检测AKP活性、硫酸亚铁法测定菌体对磷的代谢、氧化还原反应测定呼吸链脱氢酶活性、0.3%固体培养基检测Psa泳动能力；扫描电镜和透射电镜观察Psa在MIC处理下的形态变化。【结果】 5种ESB浓度作用下的抑菌圈直径均在16.00 mm以上，MIC和MBC均为2.4 mg/mL。ESB能抑制Psa生长，破坏细胞膜和细胞壁，造成电解质、蛋白质外泄，干扰菌体正常的物质和能量代谢； ESB可显著抑制Psa泳动能力，阻碍细胞的迁移。扫描电镜和透射电镜观察发现，ESB作用后的Psa菌体形态出现缩短、凹陷、畸形、破裂，细胞质外泄，菌体大量裂解等现象。【结论】ESB可破坏Psa的细胞结构，干扰物质和能量代谢，抑制菌体生长，具有开发成植物源杀菌剂的潜力。
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| 关 键 词： | 黄芩 乙醇提取物 猕猴桃溃疡病致病菌 抑菌机理 |
| 收稿时间： | 2021-11-21 |
Primary antibacterial mechanism of ethanol extract of Scutellaria baicalensis against the pathogens of kiwifruit canker |
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| PEI Hu-rong,LU Ming-xiu,LONG Li,LONG Zhang-fu.Primary antibacterial mechanism of ethanol extract of Scutellaria baicalensis against the pathogens of kiwifruit canker[J].Journal of Southern Agriculture,2022,53(2):477-485. |
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| Authors: | PEI Hu-rong LU Ming-xiu LONG Li LONG Zhang-fu |
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| Affiliation: | College of Life Sciences, Sichuan University, Chengdu 610064, China |
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| Abstract: | 【Objective】To explore the antibacterial mechanism of ethanol extract of Scutellaria baicalensis(ESB) on the pathogen of kiwifruit canker against Pseudomonas syringae pv. actinidiae(Psa) and to provide a theoretical foundation for the development of new plant-derived fungicides.【Method】The sensitivity of Psa to ESB was evaluated by the agar punch method. The minimum inhibitory concentration(MIC) and the minimum bactericidal concentration(MBC) were determined by gradient dilution method in liquid medium. The effect of ESB on Psa at 1/4 MIC,1/2 MIC,3/4 MIC and MIC was determined. Growth curre was drawn by monitored optical density of the bacterial cullture. Electric conductivity of culture supernatant was measured by conductivity meter. Extracellular soluble protein content was determined by the Coomassie brilliant blue method. Alkaline phosphatase(AKP) activity was measured with an AKP kit.Phosphorus metabolism was detected by the ferrous sulfate method. Respiratory chain dehydrogenase activity was detected by oxidationreduction reaction. Swimming motility ability was detected in 0.3% solid medium. Scanning electron microscope(SEM) and transmission electron microscope(TEM) were performed to observe Psa morphology at MIC.【Result】The inhibition zone diameter of ESB against Psa was above 16.00 mm. The MIC and MBC were 2.4 mg/mL. ESB inhibited the growth of Psa,damaged the cell membrane and cell wall which caused the leakage of proteins and electrolytes. Moreover,the ESB interfered with normal material and energy metabolism. ESB can significantly resist the swimming of Psa and hinder cell migration. ESB can significantly resist the swimming of Psa and hinder cell migration. Scanning electron microscopy and transmission electron microscopy showed that morphology of Psa was shortened,depressed,deformed and lysed with ESB,which caused the leak of cytoplasm and bacterial lysis.【Conclusion】ESB inhibit the normal growth of Psa by destroying the structure of Psa and interfering with substances and energy metabolism. ESB has the potential to be developed as a plant-derived fungicide. |
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