摘要
为研究油橄榄叶提取物对灰葡萄孢(Botrytis cinerea)的抑制作用及机理,本试验采用溶剂浸提法制备油橄榄叶提取物,再与灰葡萄孢作用,通过测定菌落直径、菌丝生物量探究提取物对灰葡萄孢菌体生长的抑制作用,通过测定相对电导率、核酸和蛋白质泄露量考察油橄榄叶提取物对菌体细胞膜完整性的影响;并基于平板培养体系,采用碘化丙啶(PI)染色法研究提取物对灰葡萄孢细胞膜通透性、活性氧(ROS)代谢的影响。结果表明,灰葡萄孢对油橄榄叶90%乙醇提取物具有高敏感性,经油橄榄叶提取物处理后其菌落直径和菌丝生物量分别为对照的20%、8.91%,膜电导率、核酸以及蛋白质泄漏量显著增加;孢内超氧阴离子(O_(2)^(·-))、过氧化氢(H_(2)O_(2))含量显著增加,还原型辅酶Ⅱ氧化酶(NOX)、过氧化氢酶(CAT)、过氧化物酶(POD)活性上升,超氧化物歧化酶(SOD)、谷胱甘肽还原酶(GR)、抗坏血酸过氧化物酶(APX)活性下降。综上,油橄榄叶90%乙醇提取物处理后的灰葡萄孢细胞膜受到损伤,导致细胞内容物外泄,并诱导ROS在胞内积累,加剧了膜脂质过氧化,使丙二醛(MDA)含量增加,进而抑制了菌体的生长。本研究结果为油橄榄叶提取物在果蔬杀菌剂中的应用提供了理论依据。
In order to explore the inhibitory effects of olive leaf extracts on B.cinerea and its mechanism,the olive extracts were obtained by solvent extraction,which were then were treated with B.cinerea,Colony diameter and mycelial biomass were detected to explore the inhibitory effect of the extracts on the growth of B.cinerea.The effects of olive extracts on cell membrane integrity were investigated by measuring relative conductivity,nucleic acid and protein leakage.The effects of extracts on cell membrane permeability and ROS metabolism of B.cinerea were studied by propidium iodide(PI)staining and plate culture system.The results showed that B.cinerea was highly sensitive to 90% ethanol extract,and the colony diameter and mycelial biomass of treated B.cinerea were 20% and 8.91% of the control.Extract treatment also significantly enhanced the levels of membrane conductivity,nucleic acid and protein leakage,and improved the content of O_(2)^(·-)and H_(2)O_(2)in the spores.The activities of enzymes related to ROS metabolism including NOX,CAT and POD were increased,and the activities of SOD,GR and APX decreased significantly.It was concluded that the cell membrane of B.cinerea treated with the extracts was damaged,resulting in the leakage of content and the accumulation of ROS in cells,which aggravated the occurrence of membrane lipid peroxidation,increased the content of MDA,and further inhibited the growth of cells.This study provides a possible way to utilize olive leaf extracts in the fruit and vegetable fungicides.
作者
田立鹏
万强贵
王婷
李春爱
蔡梦
蒲陆梅
TIAN Lipeng;WAN Qianggui;WANG Ting;LI Chun'ai;CAI Meng;PU Lumei(College of Science,Gansu Agricultural University,Lanzhou,Gansu 730070)
出处
《核农学报》
CAS
CSCD
北大核心
2023年第2期329-337,共9页
Journal of Nuclear Agricultural Sciences
基金
国家自然科学基金项目(31860472)。
关键词
油橄榄叶
灰葡萄孢
抑菌作用
活性氧代谢
olive leaves
B.cinerea
antibacterial activity
reactive oxygen metabolism
