• Korean Journal of Plant Tissue Culture
• /
• v.22 no.6
• /
• pp.311-316
• /
• 1995

In an attempt to obtain intersubgeneric somatic hybrids of Pelargonium aridum and P.zonale, protoplast isolated from the two species were fused by using polyethylene glycol(PEG) and electorfusion methods. Protoplast were isolated from cotyledon and leaf tissues using MS medium containing 550 mM sucrose, 0.7% cellulase (Onozuka R-10) and 0.4% Macerozyme. The optimum number of protoplasts per mL of culture medium was 6 x 10$^4$. Protoplast fused by the electrofusion method were more active than by PEG method. Heterokaryotically fused protoplasts formed calli when cultured in MS medium containing 550 mM glucose, 1 to 2 mg/L NAA and 0.5 to 1 mg/L BA.

• Applied Biological Chemistry
• /
• v.36 no.6
• /
• pp.557-561
• /
• 1993

The neomycin phosphotransferase II gene (nptII) was introduced into geranium (Pelargonium zonale hybrids) protoplast by using PEG or electroporation method. The presence of the introduced DNA in the protoplast and the expressions of the gene in the transformed cells were examined. The presence of the nptII DNA in the protoplasts were detected by polymerase chain reaction. The expressions of nptII gene in the transformed cells were confirmed by the nptII assay.

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.1
• /
• pp.29-34
• /
• 1994

The possibility that DNA could move out of the single cells isolated from geranium (Pelargonium zonale hybrids, 'PintoScarlet') callus was determined by the elechophoretic DNA analysis after treatment of low pH, various concentrations of KNO$_3$, 2,4-D, and GA$_3$ followed by the centrifugal force, all of which are hewn to and the physico-chemical properties of the cell wall. The centrifugal force of l,800 xg was need for DNA migration after the above treatment, but 7k300 xg was required without the treatments. In this experiment the optimum concentration (300 mg/L) of sodium dodecyl sulfate (SDS) used as an anion detergent to collect the negatively charged DNA was very critical not to damage the cell wall It can be concluded that the centrifugal force played a key role for the DNA migration through the cell wall, and the treatments of low pH (4.0), 0.5% KNO$_3$, 1.5 mg/L GA$_3$and 1mg/L 2,4-D further increased the DNA migration.

• Applied Biological Chemistry
• /
• v.36 no.6
• /
• pp.562-566
• /
• 1993

The conditions required for plant transformation through the electroporation system and for the electrofusion of the prtoplasts were investigated for geranium (Pelargonium zonale hybrids). The optimum condition for electroporation was 1.77 kV/cm for $40\;{\mu}sec$ under which 70% of the protoplasts were viable and 58% of the viable protoplasts were stained with methylene blue. The pBin19 DNA plasmid used as a carrier vector was isolated from E.coli $DH5{\alpha}$ strain, purified, identified by the electrophoresis on agarose gel and electroporated into the protoplasts. The KM8 liquid medium gave better cell division than any other media. One MHz of AC frequency with 40 V/cm of amplitude for 15 sec followed by 0.5 kV/cm of DC amplitude for $60\;{\mu}sec$ was most efficient for the electrofusion of protoplasts.