麥類와 豆類의 原形質體 融合

The fusion of protoplast of barley and bean species

The high frequency of heterokaryons is induced between soybean and barley species by using improved methods of protoplast fusion. Cell wall regeneration and incorporation of¹⁴C - glucose to protoplast in the early stage of culture was observed. The most heteroplasmic fusions occurred in the presence of 30% PEG and 10% dextran among the treated polysaccharides with 7 mM Ca²⁺ and application of centrifugal method than slide glass and micro syringe method. By treatment of 1 ul A₂C and 10 ul DMSO as fusion promoting agents, the protoplast fusion was promoted to 10% and to 80%, respectively. Over 95% of the fused protoplast could be obtained in 15 min. after fusogenics treatment for fusion between protoplasts from different species, but it began to be decreased after 1hr. Cell wall reformation was observed from protoplast and heteroplasmics within 5 days of culture of protoplast after isolation and aggregation from each crops. The maximum binding of ¹⁴C - glucose to cells occurred within 2-3days in individual protoplasts and after 5 days in fused protoplasts in culture.