摘要
日本榧原生质体的分离与纯化对于建立日本榧悬浮细胞无性系,从而获得次生代谢产物榧黄素等具有重要的经济价值。以日本榧一年生以及当年生叶片为分离材料,采用酶解法成功获得了完整的原生质体。试验结果表明,日本榧叶片细胞原生质体获得的最适酶浓度为:CPW+纤维素酶(Cellulase R-10)3.0%-6.0%、离析酶(Macerozyme R-10)5.0%-6.0%。原生质体的纯化使用蔗糖浓度为30%-44%。日本榧原生质体在0.7 mol/L甘露醇+50 mmol/L MES+0.5%PVP等渗液中做连续培养可以保持完整的细胞形态。
Isolation and purification of protoplast cell from the leaves of T.uucifera may exert great economic value for further suspension construction of somatic asexual cell and obtain secondary metabolites-kayaflavone. Enzyme methods was used to isolate the cell of T.nucifera leaf as materials successfully obtained protoplasts. The results showed that the suitable concentration for protoplast cells were ： CPW＋Cellulase （ R-10 ） 3.0%-6.0% and Macerozyme （ R-10 ） 5.0%-6.0%. The concentration 30%-44% sucrose obtained was employed for cell centrifugation in protoplast purification. The purified protoplast could keep on well-growth status for sustainable culture in the medium with 0.7 mol/L mannitol＋50 mmol/L MES＋0.5% PVP.
出处
《生物技术通报》
CAS
CSCD
北大核心
2013年第11期75-78,共4页
Biotechnology Bulletin
基金
国家自然科学基金项目(31071676)
关键词
日本榧
原生质体分离
纯化
Torreya nucifera Protoplasts Isolation Purification
