Identification of fungal pathogens associated with grapevine trunk using fluorescent-labelled ribosomal DNA probe |
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| Authors: | Mónica Fernández-González Pedro Miguel Izquierdo Cañas |
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| Institution: | 1. Departamento de Química Analítica y Tecnología de Alimentos, Instituto de Investigación Científica Aplicada de Castilla La Mancha (IRICA), Universidad de Castilla La Mancha, Ciudad Real, Spain;2. Fundación Parque Científico y Tecnológico de Castilla La Mancha, Albacete, Spain;3. Fundación Parque Científico y Tecnológico de Castilla La Mancha, Albacete, Spain;4. Departamento de Investigación, Instituto de la Vid y el Vino de Castilla La Mancha (IVICAM-IRIAF), Ciudad Real, Spain |
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| Abstract: | Detection and identification of fungal pathogens associated with grapevine trunk diseases (GTDs) are often difficult and laborious. The aim of the study is to develop a simple and time-saving protocol for the identification of a broad range of fungal species causing GTDs by fragment length analysis of the internal transcribed spacer (ITS) regions 1 and 2 and the large subunit D2 hypervariable region of the ribosomal DNA (rDNA). A total of 32 fungal isolates from declining vines and six type strains, representing 19 different species was included in this study. The majority of the fungi had unique species-specific PCR products ranging from 212 to 370 bp (ITS1), 332 to 428 bp (ITS2) and 320 to 325 bp (LSU-D2), and the combination of three regions identifies all pathogenic fungi tested. The protocol proposed here provides a highly sensitive, reliable and rapid identification method for a broad range of significant grapevine fungal trunk pathogens. |
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| Keywords: | Grapevine pathogenic fungi trunk diseases fragment lengths analysis ribosomal DNA |
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