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大花红景天再生体系的优化及抗性筛选
引用本文:王莉莎,蔡翠萍,常 凯,廖志华,兰小中. 大花红景天再生体系的优化及抗性筛选[J]. 中草药, 2014, 45(4): 558-562
作者姓名:王莉莎  蔡翠萍  常 凯  廖志华  兰小中
作者单位:1.西藏大学农牧学院,西藏 林芝 860000 2.西南大学 天然产物与代谢工程实验室,重庆 400715 3.重庆市甘薯工程技术研究中心,重庆 400715 4.成都军区总医院 临床实验医学研究与保障中心,四川 成都 610083
基金项目:国家科技支撑计划(2011BAI13B06);国家中医药管理局公共卫生专项(20120716-540000)
摘    要:目的 优化大花红景天再生体系并建立抗性筛选最佳条件,为建立大花红景天高效遗传转化体系奠定基础。方法 以大花红景天叶片为外植体,观察再生过程各阶段在不同配比的6-BA、NAA、IBA诱导下的诱导率及生长状况,并利用梯度筛选出外植体对卡那霉素(Kan)和抗潮霉素(Hyg)的抗性。结果 MS+3.0 mg/L 6-BA+1.0 mg/L NAA+700 mg/L L-Pro为叶片不定芽分化的最佳培养基,分化率达到92%;MS+700 mg/L L-Pro为根培养基;200 mg/L Kan、10 mg/L Hyg为大花红景天遗传转化的最佳筛选压;培养过程中添加10 mg/L Vc能有效抑制酚类物质的外泌。结论 优化了大花红景天植株再生体系,筛选出适宜于大花红景天遗传转化体系的Kan和Hyg筛选压。

关 键 词:大花红景天;再生体系;褐化;抗性筛选;遗传转化

Optimization of regeneration system and resistance screening of Rhodiola crenulata
WANG Li-sh,CAI Cui-ping,CHANG Kai,LIAO Zhi-hu,LAN Xiao-zhong. Optimization of regeneration system and resistance screening of Rhodiola crenulata[J]. Chinese Traditional and Herbal Drugs, 2014, 45(4): 558-562
Authors:WANG Li-sh  CAI Cui-ping  CHANG Kai  LIAO Zhi-hu  LAN Xiao-zhong
Affiliation:1.Agricultural and Animal Husbandry College of Tibet University, Linzhi 860000, China 2.Laboratory of Natural Products and Metabolic Engineering, Southwest University, Chongqing 400715, China 3.School of Life Sciences, Chongqing Engineering & Technology, Research Center for Sweetpotato, Chongqing 400715, China 4.Center of Clinical Laboratory Medicine Research and Services, General Hospital of Chengdu Military Command, Chengdu 610083, China
Abstract:Objective To optimize the regeneration system of Rhodiola crenulata, to establish the optimal conditions for resistance screening, and to lay the foundation for the establishment of the efficient genetic transformation system of R. crenulata. Methods The leaves of Rhodiola crenulata were used as explants. The influences of different ratios of 6-BA, NAA, and IAA in the medium on callus induction and growth conditions were observed, and the implant resistance of Kanamycin (Kan) and hygromycin (Hyg) was screened by gradient method. Results MS + 3.0 mg/L 6-BA + 1.0 mg/L NAA + 700 mg/L L-Pro was the optimal medium for the differentiation of leaf adventitious bud and the differentiation ratio of adventitious bud reached 92%; MS + 700 mg/L L-Pro was the medium for adventitious roots; The best selection of genetic transformation system for R. crenulata was 200 mg/L Kan and 10 mg/L Hyg; Adding 10 mg/L Vc could effectively inhibit the phenolic substances secretion. Conclusion The regeneration system of R. crenulata, is optimized, and the pressure of Kan and Hyg for genetic transformation system of R. crenulata is screened.
Keywords:Rhodiola crenulata (Hook. f. & Thomson) H. Ohba   regeneration system   browning   resistance screening   genetic transformation
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