摘要
以新疆杨（Populus alba L.var.pyramidalis）无菌苗叶片为材料,就其原生质体游离、纯化方法及影响因素等进行了研究.结果表明,cellulase R-10、hemicellulase和pectolase Y-23对原生质体产量有极显著影响.适宜新疆杨叶片原生质体游离的条件为CPW＋KM8P＋3.0%cellulase R-10＋0.5%～1.5% macerozyme R-10＋0.5%hemicellulase＋0.1%～0.5% pectolase Y-23＋0.6 mol/L甘露醇,酶解温度27℃,酶解时间8 h.在此条件下,原生质体产量达1.57×107 个/gFW,活力79.41%,而且蔗糖等密度离心法中的上浮法纯化原生质体效果最佳,最适蔗糖浓度为30%.
Protoplasts of Populus alba L. var. pyramidalis mesophyll were isolated from leaves of sterile shoot. The different factors affecting the protoplasts yield and different methods of purification were investigated. The results showed that there were highly significant differences among cellulase R-10, hemicellulase and pectolase Y-23 in the protoplasts yield. The best enzyme solution contained CPW salts, KMse medium,3.0% cellulase R-10,0. 5%-1. 5% macerozyme R-10,0. 5% hemicellulase,0. 1-0.5%pectolase Y-23,0.6 mol/L mannitol incubated at 27℃ for 8 hours. The protoplasts yield was 1.57× 10^7/gFW and the viability was 79.41% under the condition. Moreover,the best protoplasts purification method was rising method with sucrose isodensity centrifugation at concentration of 30%.
出处
《西北植物学报》
CAS
CSCD
北大核心
2007年第3期509-514,共6页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家林业局专项(2001-66)资助
关键词
新疆杨
原生质体
游离
纯化
Populus alba L. var. pyrarnidalis
protoplast
isolation
purification
