摘要
以‘石牌’广藿香无菌苗叶片为材料,对原生质体分离、纯化方法以及影响因素进行了研究。结果表明:以继代培养12～22 d的无菌苗顶芽下第3对展开叶片为材料,用0.5%果胶酶、0.2%离析酶和1.5%纤维素酶作用8 h,渗透压调节剂为11%甘露醇,‘石牌’广藿香叶肉原生质体产量达1.85×107个/g fw,活力达89%;原生质体纯化以12%聚蔗糖(Ficoll)漂浮法效果最佳。
Mesophyll protoplasts were isolated and purified from leaves of in vitro propagated Polgostemon cablin （Blanco） Benth. cv. shipaiensis, and different factors affecting protoplasts yield and activity were investigated. The results showed that young leaves of seedling subcultured for 12 - 22 days in the enzyme solution consisting of 0.5% （w/v） pectolyase Y-23, 0.2%（w/v） Macerozyme R-10 and 1. 5% （w/v） cellulase R-10, in 11% （w/v） mannitol buffered with 0.1% MES and 0.02% CaC12 for 8 h, the yield was 1.85 x 107 protoplasts per gram leaves （fresh weight）, the viability was above 89%.
出处
《亚热带植物科学》
2012年第3期25-28,共4页
Subtropical Plant Science
基金
广东省科技厅粤港关键领域重点突破项目(2009A030901011)
广州市科技计划基础研究项目(2009J1-C201)
关键词
'石牌’广藿香
原生质体分离
叶肉原生质体
Pogostem cablin cv. shipaiensis
protoplast isolation
mesophyll protoplast
