The isolation and fusion of pea and barley mesophyll protoplasts
1980
Lee, K.W. | Cho, S.H. | Cha, H.C. (Seoul National Univ., Seoul (Korea R.). Dept. of Botany)
The optimal conditions for the protoplast isolation from the leaves of pea (Pisum sativum L. cv. Sparkle) and barley (Hordeum vulgare L. cv. Baecdong) were determined in order to achieve a somatic hybridization between two species. It was revealed that the use of 0.5 M sorbitol as an osmoticum was appropriate for pea. The yield of intact protoplasts was highest (40%) when pea leaves were incubated in the enzyme solution for 4 hours. In the case of barley, the optimal concentrations oc cellulase, pectinase and mannitol as the enzyme solution were 2%, 1% and 0.35 M, respectively. The yield of barley protoplasts was highest (85%) when leaves were incubated in this enzyme solution for 3.5 hours. A fusion of protoplasts from pea and barley was induced by the PEG treatment enriched with calcium salts within 60 minutes.
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