DNA-based methodologies to detect and quantify the postharvest biocontrol agent Pantoea agglomerans CPA-2 applied on oranges
Soto Muñoz, Lourdes
Torres Sanchis, Rosario
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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 is necessary to study the traceability and survival of BCAs in their target application sites. The main objective of this work was to evaluate the persistence and quantify the population of CPA-2 after its postharvest application on orange cultivar 'Valencia Late' by molecular techniques. After application, the persistence of CPA-2 was evaluated by sampling the packing line and storage chambers, as well as on clothing of the workers by conventional PCR. The results showed that the maximum persistence of CPA-2 was lower than 3 days in surfaces of packing line. Furthermore, CPA-2 did not survive more than 1 day on working clothes, while in the environment or on different storage chamber surfaces it was not detected. In addition, the CPA-2 populations were quantified by quantitative PCR (qPCR) combined with a DNA intercalating reagent, propidium monoazide dye (qPCR-PMA) to quantify the CPA-2 viable cells on fruit surface. The qPCR-PMA method was compared with qPCR and dilution plating method. Results showed that CPA-2 populations quantiﬁed by qPCR-PMA were signiﬁcantly different compared with those obtained by qPCR during the time-course of the assay; however, no signiﬁcant differences were observed between qPCR-PMA and dilution plating. In conclusion, the persistence of CPA-2 was low at different sampling areas, suggesting that it cannot grow and survive on the surfaced sampled. Furthermore, qPCR-PMA method can be a quick and specific tool to monitor the viable population of CPA-2 on fruit surface.
Is part ofActa Horticulturae (ISHS), 2016, vol. 1144, p. 71-76
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Molecular tools applied to identify and quantify the biocontrol agent Pantoea agglomerans CPA-2 in postharvest treatments on oranges Soto Muñoz, Lourdes; Teixidó i Espasa, Neus; Usall i Rodié, Josep; Viñas Almenar, Inmaculada; Abadias i Sero, Mª Isabel; Torres Sanchis, Rosario (Elsevier B.V., 2015)Pantoea agglomerans strain CPA-2 is an effective biocontrol agent (BCA) for postharvest diseases of citrus and pome fruit. To implement their use as a control strategy is necessary to study the traceability of BCAs in the ...
Environmental monitoring of the biocontrol agent Pantoea agglomerans CPA-2 applied to citrus fruit at preharvest Soto Muñoz, Lourdes; Torres Sanchis, Rosario; Usall i Rodié, Josep; Viñas Almenar, Inmaculada; Dashevskaya, Svetlana; Teixidó i Espasa, Neus (Association of Applied Biologists, 2015)The aim of this study was to evaluate the environmental fate and behaviour of the biocontrol product based on Pantoea agglomerans CPA-2 cells after its release into the field. Three different methods, dilution plating, ...
Development of PMA real-time PCR method to quantify viable cells of Pantoea agglomerans CPA-2, an antagonist to control the major postharvest diseases on oranges Soto Muñoz, Lourdes; Teixidó i Espasa, Neus; Usall i Rodié, Josep; Viñas Almenar, Inmaculada; Crespo Sempere, Ana; Torres Sanchis, Rosario (Elsevier B.V., 2014)Dilution plating is the quantification method commonly used to estimate the population level of postharvest biocontrol agents, but this method does not permit a distinction among introduced and indigenous strains. Recently, ...