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Identification of fungal pathogens associated with grapevine trunk using fluorescent-labelled ribosomal DNA probe
- Fernández-González, Mónica, Izquierdo Cañas, Pedro Miguel
- Archiv für Phytopathologie und Pflanzenschutz 2016 v.49 no.19-20 pp. 567-574
- Vitis, fungi, internal transcribed spacers, pathogens, polymerase chain reaction, ribosomal DNA, vines
- 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.