Co-Authors:
Sikorskaite, S., Department of Agricultural Sciences, University of Helsinki, PO Box 27, FIN-00014, Finland
Vuorinen, A.L., Department of Agricultural Sciences, University of Helsinki, PO Box 27, FIN-00014, Finland
Rajamäki, M.-L., Department of Agricultural Sciences, University of Helsinki, PO Box 27, FIN-00014, Finland
Nieminen, A., The Instrument Center, University of Helsinki, PO Box 27, FIN-00014, Finland
Gaba, V., Department of Plant Pathology and Weed Science, Agricultural Research Organization, The Volcani Center, PO Box 6, Bet Dagan 50250, Israel
Valkonen, J.P.T., Department of Agricultural Sciences, University of Helsinki, PO Box 27, FIN-00014, Finland
Abstract:
Particle bombardment with a non-vacuum gene gun is an efficient method for transfection of plant cells with cloned viruses and initiation of virus infection. The HandyGun developed in this study is an improved version of a non-vacuum gene gun. Bombardment parameters were studied by inoculating an infectious, 35S promoter-driven cDNA of Potato virus A (PVA; Potyvirus) to the potato clone 'A6', Nicotiana benthamiana and N. tabacum as plasmid DNA coated on microprojectiles (gold particles). The large number of initial infection sites (necrotic local lesions) observed on inoculated 'A6' leaves and the high percentage of Nicotiana plants which were infected systemically with PVA following inoculation with HandyGun were not particularly sensitive to variation in the parameters tested (helium pressure and the amounts of plasmid DNA and gold particles). Data showed that HandyGun is a robust and reliable tool for obtaining high infection rates in plants reproducibly. It is easy and inexpensive to use and can be constructed from parts commonly available. © 2010 Elsevier B.V.
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