[关键词]
[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。
[Key word]
[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 Kan and 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 media for adventitious roots; The best selection of genetic transformation system for R. crenulata were 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, was optimized, and the pressure of Kan and Hyg for genetic transformation system of R. crenulata was screened.
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[基金项目]
国家科技支撑计划(2011BA113B06),国家中医药管理局公共卫生专项(20120716-540000)国家科技支撑计划(2011BA113B06),国家中医药管理局公共卫生专项(20120716-540000)