Biolistic inoculation of plants with Tomato yellow leaf curl virus DNA

Abstract

Tomato yellow leaf curl virus (TYLCV) full-length DNA was amplified by PCR and cloned into a bacterial plasmid. The cloned TYLCV DNA was excised from the plasmid, ligated and the resulting monomeric circular double-stranded TYLCV DNA was used to inoculate tomato (Solanum lycopersicom) and datura (Datura stramonium) plants by particle bombardment. The bombarded plants produced typical disease symptoms, similar to those produced following whitefly-mediated inoculation, albeit 5-7 days later than whitefly-inoculated plants. The success rate of inoculating tomato plants by particle bombardment averaged 37%, whereas with datura plants, it averaged 85%. With whitefly-mediated inoculation of TYLCV, the success rate of inoculation was also higher in datura plants than in tomato plants. Bombardment of datura plants with a linear form of TYLCV DNA also resulted in viral infection, with an inoculation success rate similar to that with the closed-circular TYLCV DNA. Bombarding datura plants with the bacterial plasmid containing the cloned TYLCV DNA did not result in viral infection, but bombardment with a bacterial plasmid containing a cloned dimer of TYLCV DNA yielded an infection rate of 50-100%. This is the first report of TYLCV inoculation of plants using particle bombardment of a cloned monomeric linear or closed-circular form of TYLCV double-stranded DNA.

Watermelon chlorotic stunt virus (WmCSV): a serious disease threatening watermelon production in Jordan

“…It is worth mentioning that the technique of biolistic inoculation using particle inflow gun was recently introduced into our laboratory and more experiences have to be gained to optimize the use of this technique for plant inoculation. Furthermore, several studies showed that the efficiency of plant inoculation using particle inflow gun is affected by several factors including plant age, gas pressure, the distance between the particle inflow gun and the plant as well as the source of the nucleic acid used for inoculation [24,[40][41][42]. Particle bombardment was recently used to inoculate plants with other begomoviruses: five bipartite (Bean golden yellow mosaic virus, Cabbage leaf curl virus, Tomato mottle virus, SLCV, and WmCSV) as well as one monopartite (Tomato yellow leaf curl virus) [40].…”

Infectivity of Tomato yellow leaf curl virus isolated from imported tomato fruit in Estonia

“…Agroinoculation has been an important technique for viral inoculation studies since the mid-1980s (Vaghchhipawala, Mysore, 2008), and it has successfully been used to inoculate tomato plants with cloned tandem repeats of TYLCV DNA derived from leaf tissues (Lapidot et al, 2007). Agroinoculation with the infectious 1.8-mer construct of TYLCV-[EE-Imp-05-08] amplified from tomato fruit resulted in an infectivity of 100% and the development of TYLCV symptoms both on tomato and N. benthamiana plants.…”

“…Similar inoculation efficiencies were obtained as in the current study: 50% and 75% on tomato and N. benthamiana, respectively. Lapidot et al (2007) bombarded tomato with circular full-length DNA of TYLCV with an inoculation efficiency of 37%. They also demonstrated infection after bombardment of Datura stramonium plants with a linear monomer of the TYLCV genome.…”

Biolistics for high-throughput transformation and RNA interference inDrosophila melanogaster

“…Biolistics was originally developed to deliver DNA to plants, [1][2][3][4][5][6] but the method has more recently been extended to transform a range of animals including nematodes, [7][8][9][10] crustaceans, 11 amphibians, 12 and mammalian cells and tissues. [13][14][15][16][17] Despite its apparent utility in these instances, biolistics has not yet been widely used as a method of DNA delivery for many animal species.…”