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A TaqMan-based qPCR assay for quantitative detection of the biocontrol agents Bacillus subtilis strain QST713 and Bacillus amyloliquefaciens subsp. plantarum strain D747

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Abstract

Biological control agents (BCAs) play an important role in crop protection. They can improve sustainability, prevent resistance to fungicides in target pathogens and reduce fungicide residues on produce. Bacillus subtilis strain QST713 (Serenade Max; Bayer CropScience, Leverkusen, Germany) and Bacillus amyloliquefaciens subsp. plantarum strain D747 [Amylo-X; CBC Europe S.r.l.—Biogard Division, Nova Milanese (MB), Italy] are two commercially available BCAs that are used against grey mould on table grape and other crops. TaqMan-based quantitative (q)PCR assays were developed and validated for quantitative and specific detection of the two BCAs on grape bunches following field sprays. Specific molecular markers were developed for each BCA, and TaqMan probes were designed and tested in qPCR on DNA extracted from washing water of grape berries. The assay proved specific and sensitive for both BCAs allowing detection at density as low as three colony-forming units (CFU) per gram of berries. The method will be useful to investigate the population dynamics of the two BCAs following their applications in vineyards.

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Acknowledgments

This research was carried out within the framework of the project ‘Promotion of ECO-friendly processes for the enhancement of quality of apulian food productions’ Granted by the “Distretto Agroalimentare Regionale–D.A.Re. s.c.ar.l.”.

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Authors and Affiliations

  1. Department of Soil, Plant and Food Sciences – Plant Pathology Section, University of Bari ALDO MORO, via G. Amendola 165/A, 70126, Bari, Italy

    Caterina Rotolo, Rita Milvia De Miccolis Angelini, Stefania Pollastro & Francesco Faretra

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  1. Caterina Rotolo
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  2. Rita Milvia De Miccolis Angelini
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  3. Stefania Pollastro
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  4. Francesco Faretra
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Correspondence to Rita Milvia De Miccolis Angelini.

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Handling Editor: Monica Hofte.

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Rotolo, C., De Miccolis Angelini, R.M., Pollastro, S. et al. A TaqMan-based qPCR assay for quantitative detection of the biocontrol agents Bacillus subtilis strain QST713 and Bacillus amyloliquefaciens subsp. plantarum strain D747. BioControl 61, 91–101 (2016). https://doi.org/10.1007/s10526-015-9701-4

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  • DOI: https://doi.org/10.1007/s10526-015-9701-4

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