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Hibiscus syriacus H. rosa sinensis 의 Callus 로부터 원형질체유리와 배양
정재동,조재두 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
The experiments were conducted to identify several factors influencing protoplast isolation and to determine optimal concentrations of plant growth regulators to cell division and cell colony formation through callus derived protoplast culture of Hibiscus syriacus and H. rosa sinensis. The highest yield and viability of protoplasts obtained in 8.0% cellulase combined with 0.5% macerozyme for H. syriacus, and in 6.0% of 8.0% cellulase combined with 0.5% macerozyme for H. rosa sinensis. The optimal mannitol concentration in enzyme solution was identified as 0.6M and yield of protoplast was markedly decreased in higher or lower concentration than in 0.6M. Protoplast yield of H. syriacus was continuously increased for 5 hours from initial digestion, but viability of protoplasts was maximum at 3 hours' digestion. Protoplast yield and viability of H. rosy sinensis were the highest at 4 hours' digestion, subsequently decreased. Addition of DTT to enzyme solution made protoplast yield increase, but protoplast yield was remarkably decreased by addition of DTT and citric acid in combination to enzyme solution. The highly viable and stable protoplasts were harvested by the method of E/MSSCC with 0.6M sucrose and mannitol. The useful combination of plant growth regulators with 8P-KM medium for cell division and cell colony formation from protoplast culture was identified as 2.5 ㎎/ℓ NAA, 0.3 ㎎/ℓ 2,4-D, 1.0 ㎎/ℓ BAP and 0.5 ㎎/ℓ GA.