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Young am Chae(蔡永岩),Kyu whan Choi(崔奎煥) 한국육종학회 1986 한국육종학회지 Vol.18 No.4
This study was carried out to find out the proper enzyme mix for protoplast isolation, and the effects of PEG, pH and Ca ion on the fusion. Mix of 2% cellulase R10 with 0.5% macerozyme R10 and 2.5% cellulase with 0.75% macerozyme were proper for isolation of protoplasts from mesophyll tissues of potato and egg plant, respectively. Fusion frequency was higher in each of 35% PEG(MW 6000), 15 mM Ca ion and at pH 5.5, however, their combined effect was not greater than as it was expected.
蔡永岩,崔奎煥 서울大學校 農科大學 1985 서울대농학연구지 Vol.10 No.1
Various innovations for the genetic modification of plants are being investigated in aiming to produce new varieties that can not be made by the conventional plant breeding scheme. One of the most promising approach in plant transformation is to introduce new genes into plant protoplasts and regenerate these protoplasts into whole plants with new improved characteristics. Before embark directly on a target economic crop, however, it seems a right way to establish firstly the feasibility of the protoplast approach in a model plant species where the cycle of plant to protoplast to plant is well established. When the protoplast approach was worked beautifully on the model species, then this technique would be extended to a economic crop with its difficulties in plant regeneration from the protoplasts. This report describe the procedures established through a serious of experiments for the optimum growth and regeneration conditions of the protoplasts isolated from Nicotiana plumbaginifolia leaf mesophyll cells.
崔奎煥,蔡永岩 서울大學校 農科大學 1988 서울대농학연구지 Vol.13 No.1
This experiment was carried out to determine the proper condition for protoplast isolation and fusion in Nicotiana sylvestris and N. plumbaginifolia, and plant regeneration from fused cells. Enzyme mix of 2.0% cellulase R10 0.5% macerozyme R10 was proper for protoplast isolation from leaf mesophyll cells. Optimum fusin frequency was observed in solution(pH5.8) of PEG(6000) 35% and 9mM CaCl₂Callus was induced 20 days after plating on culture medium. Shoots were formed 6 weeks after plating on shooting medium and rooted on MS basal medium thereafter. Three out of 20 regenerated plants were identfied as somatic hybrids by peroxidase isozyme analysis.