Development of a Real-time Polymerase Chain Reaction for the Detection of Fusarium oxysporum f. sp. basilici from Basil Seed and Roots |
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| Authors: | M. Pasquali P. Piatti M. L. Gullino A. Garibaldi |
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| Affiliation: | Authors' addresses: Centre of Competence for the Innovation in the Agro-environmental field (AGROINNOVA), University of Torino, Via Leonardo da Vinci 44, 10095 Grugliasco (TO), Italy;;Laboratorio Chimico della Camera di Commercio di Torino, Via Ventimiglia, 165, 10127 Torino, Italy (correspondence to M. Pasquali. E-mail: ) |
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| Abstract: | A real‐time polymerase chain reaction (PCR) assay using a TaqMan® probe was developed to detect the causal agent of wilt and crown rot of basil from infected plants and seed in Italy. The aim of the study was to diminish testing time, previously performed using nested‐PCR, and to create the conditions for future automation. The sensitivity of the assay was shown to be similar to the detection limit of the available nested‐PCR procedure. The advantages of real‐time PCR system include halving of the testing time, as well as the ability to identify both internally and externally infected seed to the sensitivity of 1 pg of genomic DNA. The assay was able to detect the presence of the pathogen in infected seed up to a sensitivity of 24 (SD: ±10) CFU per 100 seeds. |
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| Keywords: | diagnostics plant pathology prevention TaqMan® probe |
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