The study was conducted to identify several factors influencing fusion of protoplasts from mesophyll through the experiments such as PEG concentrations, DMSO dropping effect, treated period and temperature of PEG, and pH value of eluting solution,
1. Total protoplasts aggregation frequency was increased, as PEG concentration became higher, but the highest aggregation between two protoplasts occurred in 30% PEG solution.
2. DMSO dropping on PEG solution or protoplast staining with dye solution prior to PEG treatment made aggregation of two protoplasts decreased and these treatments resulted in shrivelling protoplasts.
3. The optimal incubation time in PEG solution was identified as 20 minutes, but temperature ranged 20 to 40℃ didn´t have any effects on aggregation frequency.
4. As pH of eluting solution was increased to 10.5, aggregation frequency between two protoplasts increased, but protoplasts were broken at pH 12.0. The best result was obtained between pH 9.0 arid pH 10.5.
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