东方百合‘Sorbonne’原生质体培养初步研究  

Preliminary Study on Protoplast Culture of Lilium oriental Hybrids ‘Sorbonne'

作　　者：秦晓杰[1] 段华金[1] 朱永平[2] 王小巧[1] 李琼洁[1] 赵兴富[1] 和凤美[1] 

机构地区：[1]云南农业大学园林园艺学院,昆明650201 [2]云南农业大学农学与生物技术学院,昆明650201

出　　处：《分子植物育种》2013年第5期600-604,共5页Molecular Plant Breeding

基　　金：云南省教育厅研究生项目(2011J093);云南省教育厅项目(2011J093);学生创新基金项目(NKZX2011CX023)共同资助

摘　　要：百合原生质体培养对百合远源杂交育种具有重要意义。本研究以东方百合‘Sorbonne’花托为外植体，诱导胚性愈伤组织，并对其原生质体提取和培养进行初步研究。结果表明：MS＋Picloram 3mg／L为胚性愈伤组织诱导最佳培养基；胚性愈伤组织在酶解液为2％纤维素酶R-10＋0．5％离析酶R-10＋0．05％果胶酶Y23＋147mg／L二水合氯化钙＋976mg／L2-吗啉乙磺酸＋0．6mol／L甘露醇，酶解12h，制备的原生质体产量最高达4×10^5个／mL，活性达52％；在胚性愈伤组织诱导培养基中进行原生质体的悬滴培养、液体培养、固体培养、固液结合培养和看护培养。结果表明，看护培养是最佳培养方法，并在改良MS＋Picloram 2mgm的培养基中培养2～3d观察到长细胞壁的原生质体，培养4～6d，可见原生质体的第一次分裂，培养40-45d，观察到原生质体分化成的细胞团。It is significant to culture the protoplast for wide cross breeding of Lily. In this study, the embryonic callus of receptacle of lilium oriental hybrids ＇Sorbonne＇ were induced and protoplasts were isolated and cultivated. The results showed that feasible culture medium of inductive embryonic callus was MS ＋Picloram 3 mg/L; The optimal enzyme solution used for protoplasts isolation with 12 hours enzymolysis was 2% Cellulase R-10＋ 0.5% Macerozyme R-10＋0.05% Pectolyase Y23＋147 mg/L CaCl2-2H20＋976 mg/L MES＋6 mol/L Sorbitol, and yield reached its highest, which was up to 4×10^5/mL, and the activity was up to 52%. Comparing with the hanging drop culture, liquid culture, solid culture, liquid and solid culture, the nurse culture was the best way to cultivate protoplast in feasible culture medium of inductive embryonic callus. Some changes can be observed in the process ofprotoplast culture with improved medium that is MS＋Picloram 2 mg/L by the nurse culture, such as cell wall for 2-3 days, the first division ofprotoplast for 4-6 days, cell mass for 40-45 days.

关 键 词：Sorbonne 原生质体 再生细胞 分裂 

分 类 号：S682.29[农业科学—观赏园艺]