山西农业大学学报（自然科学版）2017，Vol.37Issue(4)：254-257,4.
狭叶柴胡愈伤组织原生质体的制备及纯化
Preparation and purification of protoplasts of callus in Bupleurum scorzonerifolium Willd
摘要
Abstract
[Objective] Screening the optimal condition on callus protoplast preparation and purification of Bupleurum scorzonerifolium Willd.[Methods]The effects of some factors on the preparation and purification of protoplasts of callus were investigated,including concentration of enzyme mixture,osmotic stabilizer,incubation time and sucrose concentration.[Results]The optimal condition of protoplast isolation was:2 ％ Cellulase and 1％ Macerozyme,mannitol of 0.6 mol·L-1 as osmotic stabilizer,enzymatic hydrolysis 4 h after pre-enzymatic hydrolysis 2 h at 27℃ in dark,the protoplast yield could reach up to 1.26 × 106 per gram of callus.Under this condition,different concentration of sucrose solution was used for protoplast purification.The yield was up to 0.75 × 106 per gram of callus when purified with sucrose at the concentration of 21％.[Conclusion]In conclusion,these results lay a technological foundation of genetic engineering in B.scorzonerifolium Willd.by using protoplast.关键词
狭叶柴胡/愈伤组织/原生质体制备/纯化Key words
Bupleurum scorzonerifolium Willd./Callus/Protoplast preparation/Purification分类
生物科学引用本文复制引用
程玉鹏,李天聪,林进华,李玥玥,宁愿,马爱萍,李弘琨,王洪月..狭叶柴胡愈伤组织原生质体的制备及纯化[J].山西农业大学学报（自然科学版）,2017,37(4):254-257,4.基金项目
国家自然科学基金(81573539) （81573539）
黑龙江省自然科学基金(H2015042) （H2015042）
