Simple hand-held devices for the efficient infection of plants with viral-encoding constructs by particle bombardment.
An efficient method for infection of plants with a cloned potyvirus by particle bombardment has been described (Gal-On et al., 1995). A simplified method is described now whereby a vaccuum chamber and helium propulsive gas are not required to achieve a high efficiency of infection. The new device-the 'HandGun'--is hand-held, and easily constructed from readily available materials. With this technique it is possible to bombard soft plants and seedings that do not survive particle bombardment by other devices. bombardment of C. pepo plants with a full length clone of zucchini yellow mosaic potyvirus results in approximately 100% infection at 100 pg cDNA per plant using air or helium to propel the microprojectiles. The HandGun is 10(5)-fold more efficient than mechanical inoculation. Tungsten and gold were found to be the most efficient materials tested for use as microprojectiles. Crude extracts of plasmids from E. coli were found to be effective, as well as column-purified cDNA. A functional, simple version of the HandGun--'the Blowpipe'--was also constructed, which does not require an electrically controlled valve. Plants can be inoculated with plant viruses from sap with the HandGun.
Gal-On
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A
Department of Virology, Volcani Center, Bet Dagan, Israel. VPAMIT@VOLCANI.AGRI.GOV.IL
Meiri
E
E
Elman
C
C
Gray
D J
DJ
Gaba
V
V
D003001:Cloning, Molecular / Q000379:methods; D005821:Genetic Techniques / Q000295:instrumentation*; D017800:Potyvirus / Q000235:genetics*; D014675:Vegetables / Q000821:virology*