Elaboration of methods for detection of Pseudomonas savastanoi pv. phaseolicola on bean seeds

Abstract

Pseudomonas savastanoi pv. phaseolicola detection on artificially inoculated bean seeds was investigated. The method of the International Seed Federation - ISF (2006) was used. It includes bacteria extraction from seeds, isolation on semiselective media and checking the pathogenicity of investigated isolates. For verification of results, quick new methods of investigation were used (ELISA test and PCR). The results show that semiselective media MT (Milk Tween Agar) and MSP (Modified Sucrose Peptone Agar) can be appropriate for isolation of this bacterium. Pathogenicity of the investigated isolates was confirmed on cotyledon leaves of bean. ELISA test and PCR confirmed that all investigated isolates and reisolates belong to the bacterium P.s.pv. phaseolicola.

U radu je ispitivana mogućnost utvrđivanja prisustva bakterije Pseudomonas savastanoi pv. phaseolicola sa veštački inokulisanog semena pasulja. Korišćena je metoda International Seed Federation (ISF), koja uključuje ekstrakciju bakterija iz semena, izolaciju na poluselektivne podloge i proveru patogenosti dobijenih izolata. U cilju potvrde rezultata korišćene su brze savremene metode identifikacije (ELISA test i PCR). Rezultati dobijeni tokom rada pokazuju da su poluselektivne podloge MT (Milk Tween Agar) i MSP (Modified Sucrose Peptone Agar) pogodne za izolaciju ove bakterije. Patogenost odabranih izolata dokazana je na kotiledonim listovima pasulja. Primenom ELISA testova i PCR, potvrđeno je da svi ispitivani izolati i reizolati pripadaju bakteriji P.s.pv. phaseolicola.