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ISHS Acta Horticulturae 789: XV Meeting of the EUCARPIA Tomato Working Group

DETECTION OF PSEUDOMONAS CORRUGATA, P. MEDITERRANEA AND P. VIRIDIFLAVA IN NATURALLY INFECTED TOMATO STEMS BY BIO-PCR

Authors:   V. Passo, L. Moura, J. Duclos
Keywords:   Lycopersicon esculentum, bacteria, Tomato Pith Necrosis, TPN
Abstract:
A polymerase chain reaction (PCR) method that combines biological and enzymatic amplification (BIO-PCR) is described for the detection of fluorescent and non-fluorescent Pseudomonas, associated with tomato pith necrosis in naturally infected plants under commercial greenhouse conditions, in the north and south of Portugal. Sections from the basal, median and apical parts of tomato stems were macerated with water and aliquots of the extracts were plated onto selective agar media. After 24 h of incubation, the plates were washed to remove bacterial cells and aliquots were analyzed by PCR, without prior DNA extraction. Primers PC1/1, PC1/2, PC5/1 and PC5/2 were used in a multiplex PCR for the detection of Pseudomonas corrugata and P. mediterranea. Primers Pvir1 and Pvir2 were used in a standard PCR for the detection of P. viridiflava. BIO-PCR detected P. corrugata in the north, P. mediterranea in the south and P. viridiflava in both regions. Overall results suggested that BIO-PCR may be a useful technique for routine detection of these pathogens under field conditions, as well as for epidemiological and host-pathogen interaction studies.

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