DNA-based methodologies for the quantification of live and dead cells in formulated biocontrol products based on Pantoea agglomerans CPA-2
Soto Muñoz, Lourdes
Torres Sanchis, Rosario
Solsona Aixalà, Cristina
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Pantoea agglomerans strain CPA-2 is an effective biocontrol agent (BCA) against the major postharvest pathogens present on pome and citrus fruits. Dehydration, such as freeze-drying, spray-drying and fluidized bed drying is one of the best ways to formulate BCAs. In this work, the survival of CPA-2 cells after formulation was determined by dilution plating and molecular methods as qPCR alone and combined with a sample pretreatment with a propidium monoazide dye (PMA-qPCR) and they were used to calculate treatment concentrations in efficacy trials on postharvest oranges. Furthermore, no significant differences in CPA-2 survival were observed as determined by dilution plating and PMA-qPCR after both the freeze drying and fluidized bed drying processes; however, an interesting significant difference was observed in the spray dried product comparing all quantitative methods. A difference of 0.48 and 2.17 log10 CFU or cells g/dw was observed among PMA-qPCR with qPCR and dilution plating, respectively. According to our study, dilution plating was shown to be an unreliable tool for monitoring the survival of CPA-2 after spray drying. In contrast, the combination of PMA and qPCR enabled a quick and unequivocal methodology to enumerate viable and VBNC CPA-2 cells under stress-dried conditions. Efficacy trials showed that, after 3 days, spray drying formulation rehydrated with 10% non-fat skimmed milk (NFSM) was as effective as fresh cells to control Penicillium digitatum in oranges.
Is part ofInternational Journal of Food Microbiology, 2015, vol. 210, p. 79-83
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Detection and quantification by PCR assay of the biocontrol agent Pantoea agglomerans CPA-2 on apples Soto Muñoz, Lourdes; Teixidó i Espasa, Neus; Usall i Rodié, Josep; Viñas Almenar, Inmaculada; Torres Sanchis, Rosario (Elsevier B.V., 2014)The registration of biological control agents requires the development of monitoring systems to detect and quantify the agent in the environment Pantoea agglomerans CPA-2 is an effective biocontrol agent for postharvest ...
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DNA-based methodologies to detect and quantify the postharvest biocontrol agent Pantoea agglomerans CPA-2 applied on oranges Soto Muñoz, Lourdes; Teixidó i Espasa, Neus; Usall i Rodié, Josep; Viñas Almenar, Inmaculada; Abadias i Sero, Mª Isabel; Torres Sanchis, Rosario (International Society for Horticultural Science, 2016-11)Pantoea agglomerans strain CPA-2 is an effective biocontrol agent (BCA) for postharvest diseases of citrus and pome fruits. However, for registration purposes and to implement their use as effective control strategy, it ...