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Title

Identification and quantification of Fusarium oxysporum in planta and soil by means of an improved specific and quantitative PCR assay

AuthorsJiménez Fernández, Daniel; Montes Borrego, Miguel ; Navas Cortés, Juan Antonio ; Jiménez-Díaz, Rafael M. ; Landa, Blanca B.
KeywordsFungal DNA quantification
In planta detection
PCR specificity
Soil detection
Issue DateNov-2010
PublisherElsevier
CitationApplied Soil Ecology 46(3): 372-382 (2010)
AbstractThe proper identification and quantification of F. oxysporum populations inhabiting soil and plant rhizosphere niches are of importance for soil microbial ecology and plant pathology. In this study, we report the improvement of a PCR protocol for the specific identification of the F. oxysporum species complex and its conversion into a real-time qPCR assay for the quantification up to 1. pg of the fungus DNA in soil and different plant tissues. The amplification efficiency, sensibility and reproducibility of qPCR assays were not influenced by presence of non-target DNA from either plant or soil. The applicability of the newly developed qPCR protocol for F. oxysporum population studies was demonstrated using the technique for quantifying the fungus in different complex environmental samples. The use of the qPCR protocol allowed to accurately quantify up to 25. pg of F. oxysporum/g of naturally infested field soil, as well as to identify significant differences in the amount of F. oxysporum DNA in roots of different chickpea cultivars grown in a field soil infested with diverse pathogenic and nonpathogenic F. oxysporum populations. This qPCR protocol may be especially important for studies on soil microbial ecology and plant pathology since it provides a new opportunity for analyzing F. oxysporum populations and their interactions with the soil microflora, environment and plant host genotypes. © 2010 Elsevier B.V.
URIhttp://hdl.handle.net/10261/88660
DOI10.1016/j.apsoil.2010.10.001
Identifiersdoi: 10.1016/j.apsoil.2010.10.001
issn: 0929-1393
Appears in Collections:(IAS) Artículos
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