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DOI: 10.3852/08-085
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Mycologia, 101(4), 2009, pp. 484-495.
© 2009 by The Mycological Society of America

Identification of Indian pathogenic races of Fusarium oxysporum f. sp. ciceris with gene specific, ITS and random markers


Gayatri Gurjar 1

     Plant Molecular Biology Unit, Division of Biochemical Sciences, National Chemical Laboratory, Dr Homi Bhabha Road, Pune 411008, India

Maneesha Barve 1

     Visiting Scientist, Protein Crystallography Section, Solid State Physics Division, Bhabha Atomic Research Centre, Mumbai 400 085, India

Ashok Giri
Vidya Gupta 2

     Plant Molecular Biology Unit, Division of Biochemical Sciences, National Chemical Laboratory, Dr Homi Bhabha Road, Pune 411008, India

In this study we demonstrate the synergistic use of gene-specific markers, ITS-RFLP, ISSR and AFLP for distinguishing Indian F. oxysporum f. sp. ciceris races. We also report for the first time that F. oxysporum f. sp. ciceris race 3, a wilt pathogen of chickpea in India, is actually F. proliferatum based on phylogenetic analysis with EF-1{alpha} sequence data. F. oxysporum f. sp. ciceris races 1, 2 and 4 were easily distinguished from "race 3" (F. proliferatum) by PCR amplification with oligonucleotides designed from conserved regions of Hop78 transposon (Hop 78), cutinase (Cut), desaturase (Dst). F. oxysporum f. sp. ciceris race 4 was distinguished with the xylanase 3 (xyl3) gene by absence of amplification product only in this race. The Xyl3 amplified-DNA fragment isolated and sequenced from F. oxysporum f. sp. ciceris race 1 was similar to the F-xylanase (Xyl3) gene of F. oxysporum f. sp. lycopersici. A TELD motif, which is characteristic of the F-xylanases family, was detected within the deduced amino acid sequence of F. oxysporum f. sp. ciceris. Similarly the F. oxysporum f. sp. ciceris Hop78 DNA fragment, which identified "race 3" (F. proliferatum), was homologous to the Hop78 transposon of F. oxysporum f. sp. melonis, including the 100 amino acid conserved domain and the characteristic CCHC motif. The internal transcribed spacer region-restriction fragment length polymorphism (ITS-RFLP) approach along with intersimple sequence repeat (ISSR) method also differentiated "race 3" (F. proliferatum). Races 1 and 2 were identified by unique AFLP patterns. Sequence characterization of race-specific AFLP products revealed significant homologies of these sequences with metabolically important genes.

Key words: AFLP, F. oxysporum f. sp. ciceris races, gene specific amplification, ISSR, ITS-RFLP, race delineation, sequence characterization, translation elongation factor (EF-1{alpha})

2 Corresponding author. Tel.: +9120 25902247; fax: +9120 25902648. E-mail: vs.gupta{at}ncl.res.in






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Copyright © 2009 by The Mycological Society of America.