香豆素对灰葡萄孢霉菌的生长代谢及保护酶活性的影响

为探究香豆素对灰葡萄孢霉菌作用机理的影响，通过酶活试验、丙二醛（MDA)含量、葡萄糖含量变化试验，来分析香豆素对灰葡萄孢霉菌体内保护酶（SOD、POD、PPO）活性的影响，探究灰葡萄孢霉菌细胞代谢活动的强弱和细胞膜损伤程度变化。运用菌丝处理对照前后菌丝干重变化的方法来进行抑菌活性测定，以及用试剂盒测定的方法来进行酶活测定、MDA含量测定、葡萄糖含量的测定。结果表明，在液体培养基中香豆素对灰葡萄孢霉菌的EC₅₀为101 μg·mL^-1；香豆素对灰葡萄孢霉菌孢内的过氧化物酶（POD）、总超氧化物歧化酶（T-SOD）、多酚氧化酶（PPO）的最高抑制率分别为82.7%、66.67%、56.36%；经香豆素处理的菌丝体各时间点的丙二醛（MDA）含量明显>对照组，细胞膜透性显著增强，第72 h时，处理组MDA含量为16.53 nmol·mg^-1 prot，为对照组MDA含量的1.5倍；处理组葡萄糖含量始终<对照组，第36 h与对照组相比，处理组下降了45.63%。可见，香豆素可抑制灰葡萄孢霉菌的生长，致使细胞膜菌丝膜酯损伤，抑制灰葡萄孢霉菌酶的活性，降低细胞中葡萄糖的含量，影响细胞正常代谢生长，为灰霉病的防治提供一定的理论参考。

Effects of Coumarin on Growth Metabolism and Protective Enzyme Activities of Botrytis cinerea

In order to explore the inhibitory effect of coumarin against Botrytis cinerea,the activities of protective enzymes (SOD,POD,PPO) of B.cinerea were examined in vivo the method of enzyme activity test,measuring MDA and glucose contents to reveal the changes of metabolic activity and cell membrane permeability of B.cinerea .The method of measuring the changes of mycelial dry weight was adopted to examine the inhibitory activity.Enzyme activity,contents of MDA and glucose were measured by assay kit method.The results showed that the EC₅₀ of coumarin against B.cinerea was 101 μg/mL in liquid medium,the highest inhibitory rate of coumarin to POD,T-SOD and PPO in B.cinerea were 82.7%,66.67%,and 56.36%,respectively.The MDA contents of the mycelium treated with coumarin at each time point were significantly higher than those of the control group,and the permeability of cell membrane was significantly enhanced.At 72 h,the MDA content of the treatment group was 16.53 nmol/mg prot,1.5 times of that of the control group.The glucose content of the treatment group was always lower than that of the control group.At 36 h,compared with the control group,the glucose content of the treatment group decreased by 45.63%.It was concluded that coumarin could inhibit the growth of B.cinerea,damage the cell membrane,inhibit the activity of enzyme of B.cinerea,reduce the content of glucose in cell,and affect the normal metabolic growth of the cell.The results would provide theoretical references for the prevention and control of B.cinerea.