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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.