Development of PCR Primers for a New Fusarium oxysporum Pathogenic on Paris Daisy (Argyranthemum frutescens L.) |
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Authors: | Matias Pasquali Alberto Acquadro Virgilio Balmas Quirico Migheli Maria Lodovica Gullino Angelo Garibaldi |
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Institution: | (1) AGRINNOVA – Centre of Competence for the Innovation in the agro-environmental field, University of Torino, Via Leonardo da Vinci 44, I-10095 Grugliasco (Torino), Italy (Phone;(2) Department of Plant Protection and Center of Excellence for Biotechnology Development and Biodiversity Research, University of Sassari, Via E. De Nicola 9, I-07100 Sassari, Italy |
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Abstract: | The inverse PCR technique was applied to clone genomic DNA flanking insertion sites of sequences homologous to the transposable element Fot1 in the genome of a new pathogenic isolate of Fusarium oxysporum obtained from wilted Argyranthemum frutescens (Paris daisy). Based on the genomic flanking regions, a primer was designed which when paired to a second primer matching the Fot1 sequence allowed detection of this pathogen by PCR. The primer pair Mg5/Mg6 could specifically identify nine tested isolates of F. oxysporum from A. frutescens, when fungal genomic DNA was used as template. Moreover, the primer pair Mg5/Mg6 allowed successful detection of the pathogen in stem and root tissue from asymptomatic plants that were artificially inoculated with a representative isolate of F. oxysporum from A. frutescens. |
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Keywords: | transposons Fot1 inverse PCR molecular diagnosis |
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