Detection and quantification by PCR assay of the biocontrol agent Pantoea agglomerans CPA-2 on apples

摘要: Abstract The registration of biological control agents requires the development monitoring systems to detect and quantify agent in environment. Pantoea agglomerans CPA-2 is an effective biocontrol for postharvest diseases citrus pome fruits. semi-commercial trials was evaluated by Rodac impression plates colonies isolated were confirmed conventional PCR using SCAR primers PAGA1 PAGB1. Samples taken from different surfaces that had contact with CPA-2, surrounding environment working clothes worn handlers. Moreover, population dynamics strain determined on apple both classical plating technique real-time quantitative (qPCR). A qPCR assay a 3′-minor groove-binding (MGB) probe developed specific detection quantification P. CPA-2. Based nucleotide sequence fragment one primer set TaqMan MGB designed. SP 2 -F/SP -R showed surfaces, which verified tested against purified DNA 17 strains , 4 related species, 21 bacterial other genera whole also freshly-cut fruit vegetables. level approximately 10 3 cells per reaction, standard curve linear within range 5 log units. Results low impact. maximum persistence not longer than 5 days plastic boxes stored at 0 °C. Significant differences observed results obtained dilution plating. These may indicate presence non-degraded non-viable cells. In conclusion, novel potential tool quickly specifically monitor recent surface colonisation populations during large-scale experiments could ensure efficient successful treatments.