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In Planta and Soil Quantification of Fusarium oxysporum f. sp. ciceris and Evaluation of Fusarium Wilt Resistance in Chickpea with a Newly Developed Quantitative Polymerase Chain Reaction Assay

February 2011 , Volume 101 , Number  2
Pages  250 - 262

Daniel Jiménez-Fernández, Miguel Montes-Borrego, Rafael M. Jiménez-Díaz, Juan A. Navas-Cortés, and Blanca B. Landa

First and third authors: College of Agriculture and Forestry, University of Córdoba, Edificio C-4 ‘Celestino Mutis’, Campus de Rabanales, Ctra. Madrid-Cádiz, km 396, 14071 Córdoba, Spain; and second, fourth, and fifth authors: Institute of Sustainable Agriculture (IAS), Spanish National Research Council (CSIC), Alameda del Obispo S/N, P.O. Box 4084, 14080 Córdoba, Spain.

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Accepted for publication 22 September 2010.

Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris can be managed by risk assessment and use of resistant cultivars. A reliable method for the detection and quantification of F. oxysporum f. sp. ciceris in soil and chickpea tissues would contribute much to implementation of those disease management strategies. In this study, we developed a real-time quantitative polymerase chain reaction (q-PCR) protocol that allows quantifying F. oxysporum f. sp. ciceris DNA down to 1 pg in soil, as well as in the plant root and stem. Use of the q-PCR protocol allowed quantifying as low as 45 colony forming units of F. oxysporum f. sp. ciceris per gram of dry soil from a field plot infested with several races of the pathogen. Moreover, the q-PCR protocol clearly differentiated susceptible from resistant chickpea reactions to the pathogen at 15 days after sowing in artificially infested soil, as well as the degree of virulence between two F. oxysporum f. sp. ciceris races. Also, the protocol detected early asymptomatic root infections and distinguished significant differences in the level of resistance of 12 chickpea cultivars that grew in that same field plot infested with several races of the pathogen. Use of this protocol for fast, reliable, and cost-effective quantification of F. oxysporum f. sp. ciceris in asymptomatic chickpea tissues at early stages of the infection process can be of great value for chickpea breeders and for epidemiological studies in growth chambers, greenhouses and field-scale plots.

Additional keywords: Cicer arietinum, complete resistance, molecular markers, race-specific resistance, SYBR green.

© 2011 The American Phytopathological Society