title: Development of a fluorescent in situ hybridization (FISH) technique for visualizing CGMMV in plant tissues.
abstract: Cucumber green mottle mosaic virus (CGMMV), which belongs to the genus Tobamovirus, is a major pathogen of cucurbit crops grown indoors and in open fields. Currently, immunology (e.g., ELISA) and molecular amplification techniques (e.g., RT-PCR) are employed extensively for virus detection in plant tissues and commercial seed lots diagnostics. In this study, a fluorescent in situ hybridization (FISH) technique, using oligonucleotides whose 5'-terminals were labeled with red cyanine 3 (Cy3) or green fluorescein isothiocyanate (FITC), was developed for the visualization of the pathogen in situ. This simple and reliable method allows detection and localization of CGMMV in the vegetative and reproductive tissues of cucumber and melon. When this technique was applied in male flowers, anther tissues were found to be infected; whereas the pollen grains were found to be virus-free. These results have meaningful epidemiological implications for the management of CGMMV, particularly with regard to virus transfer via seed and the role of insects as CGMMV vectors.
authors.affiliation: Institute of Plant Pathology and Weed Research, Agricultural Research Organization, The Volcani Center, POB 6, Bet Dagan 50250, Israel. Electronic address: doritsh@volcani.agri.gov.il.
authors.forename: D
authors.initials: D
authors.lastname: Shargil
authors.affiliation: Institute of Plant Sciences, Volcani Center, Agricultural Research Organization, The Volcani Center, POB 6, Bet Dagan 50250, Israel.
authors.forename: H
authors.initials: H
authors.lastname: Zemach
authors.affiliation: Institute of Plant Sciences, Volcani Center, Agricultural Research Organization, The Volcani Center, POB 6, Bet Dagan 50250, Israel.
authors.forename: E
authors.initials: E
authors.lastname: Belausov
authors.affiliation: Institute of Plant Pathology and Weed Research, Agricultural Research Organization, The Volcani Center, POB 6, Bet Dagan 50250, Israel.
authors.forename: O
authors.initials: O
authors.lastname: Lachman
authors.affiliation: Institute of Plant Sciences, Volcani Center, Agricultural Research Organization, The Volcani Center, POB 6, Bet Dagan 50250, Israel.
authors.forename: R
authors.initials: R
authors.lastname: Kamenetsky
authors.affiliation: Institute of Plant Pathology and Weed Research, Agricultural Research Organization, The Volcani Center, POB 6, Bet Dagan 50250, Israel.
authors.forename: A
authors.initials: A
authors.lastname: Dombrovsky
mesh_terms: D018553:Cucumis sativus; D019662:Cucurbitaceae; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D017898:Tobamovirus / Q000302:isolation & purification*
keywords: Anther; CGMMV; Cucumber; Melon; Pollen grains; Tobamovirus