邻烯丙基苯酚对番茄灰霉病菌全细胞呼吸的影响

Effect of bionic fungicide 2-allylphenol on the whole cell respiration of Botrytis cinerea

利用Oxygraphy液相氧电极研究了植物源杀菌剂邻烯丙基苯酚对番茄灰霉病菌(Botrytis cinerea)全细胞呼吸的影响。结果显示,邻烯丙基苯酚处理浓度为5、10 mg/L,菌丝细胞的呼吸速率较对照分别增加了19.58%和24.56%;处理浓度为150 mg/L,可显著抑制菌丝细胞的呼吸,抑制率达到77.49%,表明邻烯丙基苯酚具有低浓度刺激而高浓度抑制灰霉病菌细胞呼吸的双重影响。用药剂处理萌芽期、非萌芽期的分生孢子,测定呼吸的结果表明,药剂对孢子的呼吸作用效果与对菌丝的相似,但以萌芽期分生孢子最为敏感。以0.01 mol/L琥珀酸钠作为病菌细胞呼吸底物时,10 mg/L邻烯丙基苯酚对病菌细胞的呼吸速率具有抑制作用,而添加0.1 mol/L葡萄糖和0.01 mol/L丙酮酸钠两种底物时却表现出刺激病菌细胞呼吸的作用,表明邻烯丙基苯酚对细胞呼吸的影响和氧化底物密切相关。反应液中单独添加呼吸电子传递链特异性位点抑制剂鱼藤酮20μmol/L、嘧菌酯10 mg/L和叠氮化钠0.01 mol/L后,病菌菌丝细胞呼吸均受到显著抑制,抑制率分别为48.94%、33.08%和39.18%。随后再添加10 mg/L邻烯丙基苯酚,不改变这种抑制效果。单独添加旁路氧化酶抑制剂水杨肟酸2 mM不影响病菌菌丝的呼吸,随后再添加10 mg/L邻烯丙基苯酚后,两者协同作用则可显著抑制菌丝的呼吸。

The effect of botanical fungicide 2-allylphenol on whole cell respiration of Botrytis cinerea was conducted by oxygraphy. When added to the phosphate buffer reaction, 2-allylphenol of 5, 10 mg/L stimulated the mycelial oxygen uptake to 19.58% and 24.56%, respectively. When 150 mg/L was added, the oxygen consumption rate decreased 77.49 % in comparison with the control. This indicated 2-allylphenol had showed a dual effect on oxygen consumption. Oxygen consumption of nongerminated and germinated conidia was also evaluated. The results were consistent with those of the mycelial oxygen uptake tests and germinated conidia showed more sensitivity. Measurements of the oxidation with three substrates glucose, succinate and pyruvic acid by whole cells revealed very striking difference. With 0.01 mol/L succinate as the substrate, the oxygen consumption of the whole cell was inhibited by 10 mg/L 2-allylphenol, while 0.1 mol/L glucose and 0.01 mol/L pyruvice acid stimulated oxygen consumption. The results revealed relativity between 2-allylphenol and the substrates. When the specific site inhibitor such as rotenone （20 μmoL/L ）, azoxystrobin （10 mg/L） or azida sodium（0.01 mol/L） were added to the phosphate buffer reaction, the oxygen consumption rate was inhibited 48.94% ,39.18% and 33.08% ,respectively. When 2-allylphenol was added then, the sensitivity of cell respiration to three inhibitors was not affected. 2mM of salicylhydroxamic acid （SHAM）, an inhibitor of alternative oxidase, had no effect on mycelium cell respiration but markedly inhibited in combination with 2-allylphenol. The results were useful to study the mode of action of 2-allylphenol.