Development of a polymerase chain reaction technique for the detection of grapevine fanleaf virus in grapevine tissue

• Published in: Phytopathology 1993-07, Vol.83 (7), p.749-753
• Main Authors: Rowhani, A, Chay, C, Golino, D.A, Falk, B.W
• Format: Article
• Language: English
• Subjects: Agronomy. Soil science and plant productions; BIOCHIMIE; Biological and medical sciences; BIOQUIMICA; Biotechnology; DIAGNOSTIC; DIAGNOSTICO; EXTRACTOS VEGETALES; EXTRAIT D'ORIGINE VEGETALE; Fundamental and applied biological sciences. Psychology; Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control; grapevine fanleaf virus; Phytopathology. Animal pests. Plant and forest protection; Plant viruses and viroids; RESISTANCE AUX MALADIES; RESISTENCIA A LA ENFERMEDAD; VARIEDADES; VARIETE; VITIS VINIFERA
• ISSN: 0031-949X; 1943-7684
• DOI: 10.1094/Phyto-83-749

A polymerase chain reaction (PCR) method has been developed to detect grapevine fanleaf virus (GFLV) in GFLV-infected grape tissue. Four sample extraction methods for infected plant tissues were compared. Although PCR could readily detect RNA in samples of GFLV RNA and virion in GFLV-infected leaf samples of Gomphrena globosa with all four extraction methods, only one method was useful for GFLV detection in grapevine tissue. Dilution of infected grape leaf samples by a 200-fold excess of healthy leaf tissue did not prevent GFLV detection by this method. Extracts from healthy grapevines prepared by methods 1, 2, and 3 prevented detection of extracted GFLV genomic RNAs by PCR, demonstrating that grape tissue extracts can inhibit either reverse-transcriptase reactions or PCR. Using method 4, GFLV could be detected in all tested cultivars of European grape, Vitis vinifera, and an American species, V. rupestris. Detection was possible in infected leaves, shoots, roots, and bark scrapings. PCR detection of as little as 128 fg of GFLV RNA was possible