A1 Refereed original research article in a scientific journal
HandyGun: An improved custom-designed, non-vacuum gene gun suitable for virus inoculation
Authors: Sikorskaite S, Vuorinen AL, Rajamaki ML, Nieminen A, Gaba V, Valkonen JPT
Publisher: ELSEVIER SCIENCE BV
Publication year: 2010
Journal: Journal of Virological Methods
Journal name in source: JOURNAL OF VIROLOGICAL METHODS
Journal acronym: J VIROL METHODS
Volume: 165
Issue: 2
First page : 320
Last page: 324
Number of pages: 5
ISSN: 0166-0934
DOI: https://doi.org/10.1016/j.jviromet.2010.02.025
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, 355 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. (C) 2010 Elsevier B.V. All rights reserved.