食品科学 ›› 2022, Vol. 43 ›› Issue (3): 213-220.doi: 10.7506/spkx1002-6630-20210305-068

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盐酸小檗碱抑制指状青霉的作用机制

林波,欧阳秋丽,余婷,郑佳佳,陶能国,李路   

  1. (湘潭大学化工学院,湖南 湘潭 411105)
  • 出版日期:2022-02-15 发布日期:2022-03-08
  • 基金资助:
    湖南省教育厅重点项目(19A476);湘潭大学博士启动基金项目(18QDZ12)

Antifungal Activity and Mechanism of Action of Berberine Hydrochloride against Penicillium digitatum

LIN Bo, OUYANG Qiuli, YU Ting, ZHENG Jiajia, TAO Nengguo, LI Lu   

  1. (College of Chemical Engineering, Xiangtan University, Xiangtan 411105, China)
  • Online:2022-02-15 Published:2022-03-08

摘要: 本实验主要对盐酸小檗碱(berberine hydrochloride,BH)抑制指状青霉(Penicillium digitatum)生长的活性以及可能的作用机制进行探讨。结果表明,BH抑制指状青霉生长的最低抑菌质量浓度(minimum inhibitory concentration,MIC)和最低杀菌质量浓度(minimum fungicidal concentration,MFC)分别为0.16 g/L和0.32 g/L;用MIC的BH处理15 h孢子萌发率仅为(4.40±1.05)%,显著低于对照组((92.48±3.35)%,P<0.05);此外,10 MFC的BH可显著延缓采后柑橘果实绿霉病的发病进程,接种5 d的果实病斑直径为(1.13±0.02)cm,显著低于对照组((5.62±0.09)cm,P<0.05);扫描电子显微镜观察结果表明,BH处理使指状青霉菌丝体褶皱变形。进一步实验发现,BH处理30 min时,指状青霉菌丝体细胞壁完整性和细胞膜通透性受损,胞外碱性磷酸酶活性和可溶性蛋白质量浓度均有所上升,但胞内ATP含量和能荷水平均降低;处理60 min时,活性氧大量积累导致膜脂过氧化,线粒体功能受损,能量代谢进一步失衡。结论:BH能有效抑制指状青霉的孢子萌发和菌丝生长,并延缓柑橘果实绿霉病的发病进程,其抑菌机理可能与影响菌丝体细胞通透性和能量代谢有关。

关键词: 盐酸小檗碱;指状青霉;柑橘果实;抑菌活性;抑菌机理

Abstract: This study was conducted to examine the antifungal activity of berberine hydrochloride (BH) on the growth of Penicillium digitatum and to elucidate the possible mechanism. The results showed that BH significantly inhibited the growth of mycelia with a minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of 0.16 g/L and 0.32 g/L, respectively. Meanwhile, the germination rate of spores was only (4.40 ± 1.05)% after 15 h of culture in the presence of BH at MIC, which was significantly lower than that recorded for the control group (92.48 ± 3.35)% (P < 0.05). BH at 10 MFC effectively delayed the development of green mold in postharvest citrus fruit. The lesion diameter was (1.13 ± 0.02) cm at five days after inoculation, which was significantly lower than that of the control group (5.62 ± 0.09) cm (P < 0.05). After BH treatment, the mycelia of P. digitatum were wrinkled and deformed as observed under a scanning electron microscope (SEM). Further experiments showed that the integrity of the mycelial cell wall and cell membrane was damaged after 30 min of BH treatment, accompanied by an increase in the activity of extracellular alkaline phosphatase and the content of soluble protein. However, the intracellular ATP content and energy charge levels decreased. After 60 min exposure, the accumulation of a large amount of reactive oxygen species (ROS) led to the peroxidation of membrane lipids, mitochondrial function damage and energy metabolism imbalance. In conclusion, BH can significantly inhibit the spore germination and the mycelial growth of P. digitatum and delay the progression of citrus green mold possibly by affecting cell permeability and energy metabolism.

Key words: berberine hydrochloride; Penicillium digitatum; citrus fruit; antifungal activity; antifungal mechanism

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