완두와 보리의 엽내세포 원형질체 분이 및 융합

The Isolation and Fusion of Pea and Barley Mesophyll Protoplasts

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 0.5M sorbitol as an osmoticum was appropriate for pea. The yield of intact protoplasts was the highest (40%) when pea leaves were incubated in the enzyme solution for 4 hours. In case of barley, the optimal concentrations of cellulase, pectinase and mannitol as the enzyme solution were 2%, 1% and 0.35M, respectively. And the yield of barley protoplasts was the highest(87%) when leaves were incubated in this enzyme solution for 3.5 hours. A fusion of protoplasts from pea and barley was induced by PEG treatment enriched with calcium salts within 60 minutes.