Comparative efficiency of chemical compounds for in vitro and in vivo activity against Clavibacter michiganensis subsp. michiganensis, the causal agent of tomato bacterial canker

Abstract

Bacterial canker of tomato caused by Clavibacter michiganensis subsp. michiganensis produces considerable economic losses in many countries because effective control measures are lacking. The extent to which bactericides control this disease effectively is low and has not yet been well documented for Southern European conditions. In this study the bactericidal effect of several products on this pathogen was assessed in vitro and in vivo in tomato plants under greenhouse conditions. Seven antibacterial substances (bronopol, copper sulphate, kasugamycin, oxolinic acid, oxytetracycline, streptomycin and 8-hydroxy-quinoline), three commercial formulates (Antibak RZ, an oligoelements mixture containing copper plus zinc; Orthopol, a potassium soap; and Param, a resistance inductor) and combinations thereof were tested. In vitro assessment showed that minimal inhibitory concentration (MIC) of antibacterial substances was between 4–8 μg ml−1, except for copper sulphate with a MIC value of 150 μg ml−1 and kasugamycin, which was not active at 500 μg ml−1. MIC values of commercial formulates ranged between 5 and 40 μl ml−1. Furthermore, combinations of 8-hydroxy-quinoline+copper sulphate, 8-hydroxy-quinoline+Antibak RZ, streptomycin+Antibak RZ and streptomycin+Orthopol showed a synergistic effect at sub-inhibitory concentrations. Treatments containing copper sulphate greatly reduced disease symptoms on plants sprayed with the bacteria, whereas streptomycin was less effective. In two independent trials, the percentage of leaves showing symptoms was significantly lower (2.4% and 11.9%) after treatment with copper sulphate combined with 8-hydroxy-quinoline at half-dose, than in inoculated controls (75.1% and 59.6%). These results were better than copper sulphate alone. However, plants inoculated by pricking rapidly developed systemic infection, which no product managed to control significantly, although several treatments did reduce symptoms. We conclude that copper sulphate combined with 8-hydroxy-quinoline may be useful in controlling external symptoms of this disease in greenhouses, and is environmentally friendly, reducing the amount of copper applied to crops.