3种植物精油主要成分及其对烟草疫霉的抑菌活性

Main components of three plant essential oils and their antimicrobial activity on Phytophthora nicotianae

  • 摘要: 【目的】 探究丁香精油、山鸡椒精油和柠檬桉精油主要成分及其对烟草疫霉的抑菌活性,为开发防治烟草黑胫病的植物源农药提供理论依据。【方法】 采用气相色谱—质谱联用(GC-MS)分析丁香精油、山鸡椒精油和柠檬桉精油的主要成分;利用菌丝生长速率法测定3种植物精油及其主要成分对烟草疫霉的抑菌活性;使用扫描电镜和透射电镜观察3种植物精油对烟草疫霉菌丝形态和亚细胞结构的影响;通过盆栽试验测定3种植物精油主要成分对烟草黑胫病的防治效果。【结果】 丁香精油的主要成分为丁香酚,相对含量为82.93%;山鸡椒精油的主要成分为柠檬醛,其中顺式柠檬醛相对含量为28.48%,反式柠檬醛相对含量为42.41%;柠檬桉精油的主要成分为香茅醛,相对含量为79.27%。3种植物精油及其主要成分对烟草疫霉均有明显的抑制作用,丁香精油、山鸡椒精油和柠檬桉精油对烟草疫霉的有效中浓度(EC50)分别为191.5、197.3和185.9 μg/mL,丁香酚、柠檬醛和香茅醛对烟草疫霉的EC50分别为111.1、128.4和141.1 μg/mL。经3种植物精油处理后,通过扫描电镜观察发现,烟草疫霉菌丝断裂、畸变,菌丝分枝增多,形成分生孢子,菌丝结节出现膨胀,内部出现空泡;通过透射电镜观察发现,烟草疫霉细胞膜和线粒体膜降解,线粒体嵴消失,产生自噬小体,细胞膜周围出现小囊泡,细胞呈凋亡状态。盆栽试验结果表明,在接种烟草疫霉第7 d,丁香酚、柠檬醛和香茅醛对烟草黑胫病的防治效果分别为67.8%、50.8%和48.8%。【结论】 丁香精油、山鸡椒精油、柠檬桉精油及其主要成分对烟草疫霉均有较好的抑菌活性,其抑菌机制可能与3种植物精油能破坏烟草疫霉的细胞结构有关;3种植物精油的主要成分丁香酚、柠檬醛和香茅醛对烟草黑胫病均有较好的防治效果,具有开发成植物源农药的潜力。

     

    Abstract: 【Objective】 This study aimed to investigate the main components of essential oils of Eugenia caryophyllata Litsea cubeba and Eucalyptus citriodora, and their antimicrobial activities against Phytophthora nicotianae, so as to provide a theoretical basis for developing plant-derived pesticides for controlling tobacco black shank.【Method】 Gas chromatography-mass spectrometry (GC-MS) was used to analyze the main components of the essential oils of Eugenia caryophyllataLitsea cubeba and Eucalyptus citriodora. Mycelial growth rate assay was applied to investigate antimicrobial activities of the three plant essential oils and their main components against Phytophthora nicotianae. Scanning electron microscope (SEM) and transmission electron microscope (TEM) were used to observe effects of the three plant essential oils on the hyphal morphology and subcellular structures of Phytophthora nicotianae. Control effects of the main components of the three plant essential oils against tobacco black shank was investigated in a pot experiment.【Result】 The main component of Eugenia caryophyllata essential oil was eugenol, with its relative content of 82.93%; the main component of Litsea cubeba essential oil was citral, and the relative content of cis-citral and trans-citral were 28.48% and 42.41%; the main component of Eucalyptus citriodora essential oil was citronellal, with its relative content of 79.27%. The three plant essential oils and their main components showed obvious inhibitory effects on Phytophthora nicotianae: the median effective concentration (EC50) values of essential oils of Eugenia caryophyllataLitsea cubeba, and Eucalyptus citriodora on Phytophthora nicotianae were 191.5, 197.3, and 185.9 μg/mL, respectively; the EC50 values of eugenol, citral, and eugenol on Phytophthora nicotianae were 111.1, 128.4, and 141.1 μg/mL, respectively. After treatments of three plant essential oils, SEM observation found that Phytophthora nicotianae hyphae exhibited fragmentation and deformation, with increased branching forming conidia, and the hyphal knots became swollen, with vacuoles appearing internally. TEM observation found that the cell membrane and mitochondria membrane of Phytophthora nicotianae degraded, with disappeared mitochondrial cristae and generated autophagic vesicles, as the cells underwent apoptosis. The pot experiment results showed that on 7 d of Phytophthora nicotianae inoculation eugenol, citral, and citronellal exhi-bited control effects on tobacco black shank of 67.8%, 50.8%, and 48.8%, respectively.【Conclusion】 Essential oils of Eugenia caryophyllata Litsea cubeba and Eucalyptus citriodora show favourable antimicrobial activities against Phytophthora nicotianae, and their antimicrobial mechanism may relate to the ability of these three plant essential oils to disrupt the cellular structure of Phytophthora nicotianae. The principal components of these three plant essential oils—eugenol, citral, and citronellal—demonstrate effective control against tobacco black shank, suggesting their potential to be deve-loped as plant-derived pesticides.

     

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