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Molecular detection of Fusarium oxysporum f. sp. vasinfectum in cotton roots by PCR andreal-time PCR assay

Abd-Elsalam, Kamel A. and Schnieder, Frank and Migheli, Quirico and Verreet, Joseph Alexander (2006) Molecular detection of Fusarium oxysporum f. sp. vasinfectum in cotton roots by PCR andreal-time PCR assay. The Journal of Plant Diseases and Protection, Vol. 113 (1), p. 14-19. ISSN 1861-3829. Article.

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Abstract

A PCR assay based on a pair of oligonucleotide primers targeting the 16S and 23S rRNA genes was used to detect Fusarium oxy­sporum f. sp. vasinfectum (Fov), a fungus causing Fusarium wilt of cotton, in infected cotton seedlings. By using the primer pair Fov1-Eg-f and Fov1-Eg-r, a 438-bp DNA fragment was consistently amplified from 41 Egyptian isolates of Fov (race 3), while no amplification was obtained from template genomic DNAs obtained from other fungal pathogens, namely F. oxysporum, F. solani, F. verticillioides, F. sambucinum, Rhizoctonia solani, and Macrophomina phaseolina. PCR product of the expected size was amplified from as little as 100 fg of purified Fov genomic DNA. The sensitivity of real-time PCR allowed detection of 200 fg of target genomic DNA. The melting temperature (Tm) values of Fov isolates were in the range 86.83 to 87.02. Fov DNA was detected in infected cotton roots, while no amplification was obtained from other plant structures, such as stem and leaf. The real-time PCR assay successfully detected Fov and correlated well with assessments based on conventional PCR techniques. The application of this technique will be useful for identification of Fov in infected cotton roots and detection of Fov in infected plants that lack visible morphological structures or symptoms.

Item Type:Article
ID Code:729
Status:Published
Refereed:Yes
Uncontrolled Keywords:Cotton, Fusarium wilt, 16S rDNA, real-time PCR, species-specific primers
Subjects:Area 07 - Scienze agrarie e veterinarie > AGR/12 Patologia vegetale
Divisions:001 Università di Sassari > 02 Centri > Centro di eccellenza interdisc. sviluppo ricerca biotecnologica e studio biodiversità della Sardegna e dell'area mediterranea
001 Università di Sassari > 01 Dipartimenti > Protezione delle piante
Publisher:Eugen Ulmer
ISSN:1861-3829
Additional Information:Già Zeitschrift für Pflanzenkrankheiten und Pflanzenschutz.
Deposited On:18 Aug 2009 10:03

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