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Detection of Fusarium oxysporum f. sp. dianthi in carnation tissue by PCR amplification of transposon insertions

Chiocchetti, Annalisa and Bernardo, Ilaria and Daboussi, Marie-Josée and Garibaldi, Angelo and Gullino, Maria Lodovica and Langin, Thierry and Migheli, Quirico (1999) Detection of Fusarium oxysporum f. sp. dianthi in carnation tissue by PCR amplification of transposon insertions. Phytopathology, Vol. 89 (12), p. 1169-1175. ISSN 0031-949X. Article.

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DOI: 10.1094/PHYTO.1999.89.12.1169

Abstract

Strains of the carnation wilt pathogen, Fusarium oxysporum f. sp. dianthi, can be distinguished by DNA fingerprint patterns, using the fungal transposable elements Fot1 and impala as probes for Southern hybridization. The DNA fingerprints correspond to three groups of F. oxysporum f. sp. dianthi strains: the first group includes isolates of races 1 and 8; the second group includes isolates of races 2, 5 and 6; and the third group includes isolates of race 4. Genomic DNAs flanking race-associated insertion sites of Fot1 (from races 1, 2, and 8) or impala (from race 4) were amplified by the inverse polymerase chain reaction (PCR) technique. These regions were cloned and sequenced, and three sets of primers overlapping the 3' or 5' end of the transposon and its genomic insertion were designed. Using fungal genomic DNA as template in PCR experiments, primer pairs generated amplification products of 295, 564 and 1,315 bp, corresponding to races 1 and 8; races 2, 5, and 6; and race 4, respectively. When multiplex PCR was performed with genomic DNA belonging to races 1 and 8, 2, or 4, single amplimers were generated, allowing clear race determination of the isolate tested. PCR was successfully performed on DNA extracted from susceptible carnation cv. Indios infected with isolates representative of races 1, 2, 4, and 8.

Item Type:Article
ID Code:67
Status:Published
Refereed:Yes
Uncontrolled Keywords:Dianthus caryophyllus, Fusarium wilt, physiological race
Subjects:Area 07 - Scienze agrarie e veterinarie > AGR/12 Patologia vegetale
Divisions:001 Università di Sassari > 01 Dipartimenti > Protezione delle piante
Publisher:The American Phytopathological Society
ISSN:0031-949X
Deposited On:18 Aug 2009 10:01

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