摘要
以4℃低温暗处理24 h的南蛇藤胚性愈伤组织为分离原生质体的原材料,用MS培养基进行液体浅层静置、固液双层以及琼脂糖包埋培养原生质体,获得再生愈伤组织并分化成苗,建立了原生质体培养体系。结果表明,低温暗处理利于高产率高质量原生质体的获得;0.5%纤维素酶+0.5%果胶酶+5 mmol.L-1MES为酶的最佳配方;12 h为最佳酶解时间;13%为甘露醇最佳浓度;静置12 h+振荡0.5 h为最佳酶解方式;液体浅层静置培养取得了较好的原生质体培养效果;MS+6-BA2.0 mg.L-1+IBA 0.1 mg.L-1为愈伤组织最佳分化培养基;1/2MS+NAA0.1 mg.L-1为最佳生根培养基。
Experimental raw material of protoplast was obtained from embryo callus induction of Celastrus orbiculatus Thunb.under the condition of lower temperature at 4℃ and dark treatment for 24h.The created culture system of protoplast originated from seedlings differentiation of callus regeneration after the culture of settling the shallow-layer solution and solid-liquid double layer culture as well as agarose-embedding based on MS medium.The results showed that： the lower temperature and dark treatment was beneficial to obtain protoplast with high yield and top quality;the optimum combination for the enzyme activity would be： 0.5% Cellulase＋0.5% Pectinase＋5mg·L-1 MES;the optimum time of enzymatic hydrolysis was 12h;the optimum mannitol concentration was 13%;moreover,the optimum mode of enzymatic hydrolysis would be： standing for 12h and oscillating for 0.5h;meanwhile,better protoplast culture efficiency was based on the culture of settling the shallow-layer solution;the optimum differentiation medium of callus was MS＋6-BA 2.0mg·L-1＋IBA 0.1mg·L-1 and rooting medium could be 1/2 MS＋NAA 0.1mg·L-1.
出处
《植物研究》
CAS
CSCD
北大核心
2011年第3期300-305,共6页
Bulletin of Botanical Research
基金
国家自然科学基金(30960064)
甘肃省自然科学研究基金(1010RJZA027)
甘肃省教育厅研究生导师科研项目(0810-02)
甘肃省庆阳市科技攻关计划项目(1003NKCM067)
关键词
南蛇藤
愈伤组织
原生质体培养
植株再生
Celastrus orbiculatus Thunb.
callus
protoplast culture
plant regeneration
