Synergistic Control Effect of Bacillus pumilus AR03 and Fungicides Against Tobacco Bacterial Wilt
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摘要: 为探究短小芽孢杆菌与化学杀菌剂联合防控烟草青枯病的可行性，分别采用改良抑菌圈法和平板菌落计数法测定7种杀菌剂和短小芽孢杆菌AR03对青枯雷尔氏菌的毒力及杀菌剂与AR03的生物相容性，同时采用Horsfall法确定杀菌剂和AR03的复配比例。室内毒力试验结果表明，7种杀菌剂和AR03对青枯雷尔氏菌的生长均有较好的抑制作用。7种杀菌剂的毒力大小依次为三氯异氰尿酸、氯尿·硫酸铜、噻菌铜、溴菌·壬菌铜、甲霜·福美双、噻唑锌和中生菌素，EC50值介于101.02~212.70 mg/L之间。浓度为1.0×105~1.0×109 cfu/mL的AR03对青枯雷尔氏菌的抑菌率介于26.13%~73.54%之间，呈现浓度依赖性。生物相容性分析发现7种供试药剂与AR03的生物相容性差异较大，短小芽孢杆菌AR03与噻菌铜、噻唑锌、甲霜·福美双生物相容性较好，尤其是噻菌铜表现最好，测试浓度100 mg/L时，菌落数大于1×107cfu/mL。综合杀菌剂对青枯病菌的毒力及其与AR03生物相容性，噻菌铜表现最优。噻菌铜（EC50=175.21 mg/L）与AR03（EC50=6.84×106 cfu/mL）复配剂在体积比为5∶5时，对青枯雷尔氏菌的抑制效果显著，增效作用明显，增效比率值IR值为1.482。室内盆栽试验结果表明，菌药复配剂的防效（68.77%）明显优于单剂噻菌铜和生防菌AR03的防效，且混配剂中噻菌铜使用量只有单剂的1/2，大幅降低了化学药剂的使用量。Abstract: To explore the potential of the control synergism of B. pumilus AR03 combined with seven tested fungicides against tobacco bacterial wilt, the toxicity of seven fungicides and AR03 strain against Ralstonia.solanacearum and the bio-compatibility of fungicides with AR03 were measured by the improved inhibition zone method and the plate colony-counting method, and the synergistic effect was determined by the Horsfall method. The results of in vitro toxicity test indicated that seven fungicides and AR03 all exhibited a good inhibitory effect on the growth of R. solanacearum, and the virulence of seven fungicides were trichloroiso cyanuric acid>chloroisobromine cyanuric acid plus copper sulfate>thiodiazole copper>bromothalonil plus cuppric nonyl phenolsulfonate> Metalaxyl plus thiram> thiadiazole zinc>Zhongshengmycin, and the half-maximal inhibitor concentration (EC50) ranged from 101.02-212.70 mg/L. B. pumilus AR03 also had a good inhibition rate (26.13%-73.54%) in a dose dependent manner at the concentration from 1.0×105 to 1.0×109 cfu/mL. Results from the bio-compatibility test showed that AR03 had good biocompatibility with thiodiazole copper, Metalaxyl plus thiram, thiadiazole zinc, while thiodiazole copper performed the best. When the test concentration of 100 mg/L, the number of colonies was greater than 1×107 cfu/mL. The mixture of thiodiazole copper (EC50=175.21 mg/L) and AR03 (EC50=6.84×106 cfu/mL) in the volume ratio of 5:5 behaved a notable inhibitory and synergistic effect against R. solanacearum and yielded an IR value of 1.482. The results of in vivo experiment showed that the control effect of the combination was significantly higher than that of thiodiazole copper and AR03 strain treatment alone. More importantly, the usage volume of thiodiazole copper in the mixture was half of the single agent, which greatly reduced the use of chemical agents.
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