Study on the Inhibitory Activity and Mechanism of Athyrium sinense Essential Oil Against Clavibacter michiganensis subsp. michiganensis

doi:10.11924/j.issn.1000-6850.casb2022-0828

Abstract

Abstract:

The aim of this study was to investigate the antibacterial effect and mechanism of Athyrium sinense essential oil against Clavibacter michiganensis subsp. michiganensis (Cmm). The antibacterial effect of A. sinense essential oil on Cmm was determined by the plate punching method and the turbidimetric method. The mechanism of bacterial inhibition was mainly studied from three aspects including cell wall, cell membrane and cell cycle. The activity of extracellular alkaline phosphatase (AKP) was measured to analyze the effect on cell wall. The level of intracellular reactive oxygen species, the content of extracellular soluble protein, the potential of cell membrane, and the ATPase activity were measured to analyze the effect on cell membrane. Finally, the cell cycle was measured. The diameter of Cmm inhibition circle was (19.06±1.83) mm and the minimum inhibitory concentration (MIC) was 0.313 mg/mL, indicating that A. sinense essential oil had a strong inhibitory effect on Cmm. Analysis of the inhibition mechanism showed that under the treatment of A. sinense essential oil, extracellular AKP activity, ROS level, and extracellular soluble protein leakage increased significantly (P<0.05), cell membrane potential and intracellular ATPase activity were significantly (P<0.05) decreased, and the proportion of Cmm in DNA replication was decreased. The results showed that A. sinense essential oil had significant antibacterial effect on Cmm. The antibacterial mechanism was as follows. The destruction of Cmm cell wall led to the increase of extracellular AKP activity. The destruction of cell membrane led to a significant (P<0.05) increase in the level of reactive oxygen species and the leakage of soluble protein, the occurrence of cell membrane depolarization and the significant (P<0.05) decrease in ATPase activity. Finally, A. sinense essential oil also interfered with the normal cell cycle of Cmm.

Key words: Athyrium sinense, essential oil, Clavibacter michiganensis subsp. michiganensis, antibacterial activity, antibacterial mechanism

QU Hong, WANG Mengliang, YAN Ran, WANG Qi, CAI Jin. Study on the Inhibitory Activity and Mechanism of Athyrium sinense Essential Oil Against Clavibacter michiganensis subsp. michiganensis[J]. Chinese Agricultural Science Bulletin, 2023, 39(31): 139-146.

Figures/Tables 8

References 38

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挥发油浓度/(mg/mL)	番茄溃疡病菌	挥发油浓度/(mg/mL)	番茄溃疡病菌
5	－	0.08	++
2.5	－	0.04	+++
1.25	－	0.02	+++
0.625	－	0.01	+++
0.313	－	对照	+++
0.156	+

挥发油浓度/(mg/mL)	番茄溃疡病菌	挥发油浓度/(mg/mL)	番茄溃疡病菌
5	－	0.08	++
2.5	－	0.04	+++
1.25	－	0.02	+++
0.625	－	0.01	+++
0.313	－	对照	+++
0.156	+