title: HandGun-mediated inoculation of plants with viral pathogens for mechanistic studies.
abstract: Particle bombardment is an efficient method for virus inoculation of intact plants. This technique enables inoculation with full-length infectious clone cDNA, PCR products, virus from sap or virus preparation, and in vitro viral transcripts. The inoculation of some phloem-limited RNA and circular DNA viruses is also possible. The technique of bombardment without the use of vacuum permits the inoculation of soft-leaved plants that do not usually survive bombardment inoculation, the investigation of viral recombination in planta, promoter analysis, monitoring virus movement using an infectious clone bearing a reporter gene and the inoculation of large numbers of plants. The inoculation of whitefly-borne circular DNA begomoviruses is now possible due to direct genome amplification by Rolling Circle Amplification (RCA), followed by bombardment using a device that does not require a vacuum for operation. Here we describe the inoculation of intact plants with (a) RNA virus infective clones and (b) begomoviruses after direct genome amplification by RCA, using a handheld bombardment device.
authors.affiliation: Department of Plant Pathology and Weed Science, The Volcani Center, Agricultural Research Organization, Bet Dagan, Israel. vpgaba@volcani.agri.gov.il
authors.forename: Victor
authors.initials: V
authors.lastname: Gaba
authors.forename: Moshe
authors.initials: M
authors.lastname: Lapidot
authors.forename: Amit
authors.initials: A
authors.lastname: Gal-On
mesh_terms: D053839:Begomovirus / Q000235:genetics* / Q000502:physiology; D019470:Biolistics / Q000295:instrumentation*; D003001:Cloning, Molecular; D028464:Cucurbita / Q000235:genetics* / Q000821:virology*; D018076:DNA, Complementary / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics; D006046:Gold / Q000737:chemistry; D021141:Nucleic Acid Amplification Techniques; D017800:Potyvirus / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D014414:Tungsten / Q000737:chemistry
chemical_list: D018076:DNA, Complementary; D006046:Gold; D014414:Tungsten