Improved Detection of Petunia Vein Clearing Caulimovirus

• Published in: HortScience Vol. 35; no. 7; pp. 1279 - 1282
• Main Authors: Zeidan, M, Sikron, Noga, Cohen, J, Gera, A
• Format: Journal Article
• Language: English
• Published: Alexandria, VA American Society for Horticultural Science 01.12.2000
• Subjects: Biological and medical sciences; Biotechnology; Fundamental and applied biological sciences. Psychology; Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control; Phytopathology. Animal pests. Plant and forest protection; Plant viruses and viroids; West Asia; Israel; Asia; Mediterranean Basin; Plant pathology; Petunia hybrida; Sap; Horticulture; Plant pathogen; Process improvement; Immunological method; Petunia vein clearing virus; Dicotyledones; Angiospermae; Immunoelectron microscopy; Molecular probe; Solanaceae; Nucleic acid; Method study; Planting stock; Enzyme; Transferases; Nursery (plant); Plant leaf; Experimental study; RNA-directed RNA polymerase; Virology; Polymerase chain reaction; Nucleotidyltransferases; Transmission electron microscopy; DNA; Flower crop; Spermatophyta; Molecular biology; Detection; Cultivar
• ISSN: 0018-5345; 2327-9834
• DOI: 10.21273/hortsci.35.7.1279

Petunia vein clearing virus (PVCV), a possible member of the caulimovirus group, was detected in several cultivars of vegetatively propagated petunias ( Petunia × hybrida Hort. Volm.-Andr.) grown in commercial nurseries. Leaf dip preparations and ultrathin sections of leaf tissue were analyzed by transmission electron microscopy (TEM). Spherical virus particles, 45-50 nm in diameter, were observed in samples taken from symptomatic petunia plants. The virus was purified and a polyclonal antiserum was prepared. In immuno-specific electron microscopy (ISEM), the PVCV antiserum-treated samples reacted with a distinct decoration on the virus suspect particles. A polymerase chain reaction (PCR)-based assay was used to detect PVCV in total nucleic acid extracts derived from infected petunia plants. Two primer pairs were designed to flank a 736-base-pair sequence located in the RNA-dependent RNA polymerase gene of the PVCV genome. A DNA fragment of predicted size was visualized in agarose gels. The authenticity of the amplified DNA fragment was confirmed by restriction analysis and by hybridization with the virus-specific PVCV DNA probe. The virus could be detected efficiently in high dilutions of sap extracted from infected petunia plants .