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Development of a quantitative PCR assay for the detection of Piggotia coryli, the causal agent of hazelnut anthracnose

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Abstract

A novel real-time PCR assay was developed for the detection of Piggotia coryli, the causal agent of hazelnut anthracnose. A molecular tool for sensitive detection and quantification of P. coryli in symptomatic and asymptomatic host tissues was required to better understand P. coryli biology and epidemiology. Species-specific primers for P. coryli were designed based on the sequence of the β-tubulin gene. The amplification efficiency of pure target DNA was 93.2%, wet lab-tested limit of detection (LOD) was fixed at 8 pg/PCR reaction with maximum values of repeatability and reproducibility. In samples collected from infected orchards, the assay consistently revealed the presence of the pathogen in all symptomatic specimens, as well as in some asymptomatic tissues. This molecular tool will be of substantial aid in detecting and quantifying P. coryli, even in latent state and in different hazelnut tissues.

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Data availability

The data that support the findings of this study are available from the corresponding upon request.

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Funding

This work has been supported by the European Commission under the Grant Agreement number 774571 (project PANTHEON ‘Precision farming of hazelnut orchards’).

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Authors and Affiliations

  1. Dipartimento di Scienze Agrarie e Forestali, Università degli Studi della Tuscia, Via S. Camillo de Lellis snc, 01100, Viterbo, Italy

    Mounira Inas Drais, Silvia Turco, Chiara D’Attilia, Valerio Cristofori & Angelo Mazzaglia

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  1. Mounira Inas Drais
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  2. Silvia Turco
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  3. Chiara D’Attilia
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  4. Valerio Cristofori
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  5. Angelo Mazzaglia
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Correspondence to Angelo Mazzaglia.

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Supplementary Information

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42161_2023_1326_MOESM1_ESM.docx

Supplementary file1 Melting curve analysis from the real time PCR assay, the peaks at 85.8 °C correspond to the dissociation curve of four specific amplification products obtained (DOCX 35 KB)

Supplementary file2 List of primers used for the molecular characterization of Piggotia coryli strains (DOCX 20 KB)

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Drais, M.I., Turco, S., D’Attilia, C. et al. Development of a quantitative PCR assay for the detection of Piggotia coryli, the causal agent of hazelnut anthracnose. J Plant Pathol 105, 507–516 (2023). https://doi.org/10.1007/s42161-023-01326-z

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