黄瓜高效原生质体瞬时表达体系的建立

• 作 者: 黄洪宇[1]王振雨[2]成金桃[2]赵文超[2]李欣[2]王虹云[2]张振贤[2]睢晓蕾[2]
• 机 构: 设施蔬菜生长发育调控北京市重点实验室,北京 100193
• 出 处: 中国园艺学会黄瓜分会第四届年会
• 发表日期: 2013年期

　　The transient gene expression system using plant protoplasts has become widely used for high-throughput analysis and functional characterization of genes.In this work we investigated protoplast isolation and green fluorescent protein (GFP) transient transfection and their main affecting factors, such as mannitol concentration in enzymolysis solution, enzymolysis time, and polyethylene glycol (PEG) concentration and transfection time, on xintaimici cucumber.The results showed that when the enzyme solution had 1.5% cellulase R-10 (w/v), 0.4% macerozyme R-10 (w/v), 0.4 M mannitol, 20 mM 2-morpholinoethanesulfonic acid, 10 mM CaCl2, 0.1% bovine serum albumin, and was at pH 5.8 with an enzymolysis time of 8 h, the protoplast yield was 6-7×106 per gram fresh weight.Viability was about 90%.