大花红景天再生体系的优化及抗性筛选.pdf

大花红景天再生体系的优化及抗性筛选 1 1 2, 4 2, 3 1 王莉莎 ，蔡翠萍 ，常 凯 ，廖志华 ，兰小中 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. 中储粮成都粮食储藏科学研究所，四川 成都 610031 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 中图分类号：R282.21 文献标志码：A 文章编号：0253-2670(2014)03 - 0 - 05 DOI: 10.7501/j.issn.0253-2670.2014.03. Optimization of regeneration system and resistance screening of Rhodiola crenulata 1 1 2, 3 2, 3 1 WANG Li-sha , CAI Cui-ping , CHANG Kai , LIAO Zhi-hua , LAN Xiao-zhong 1. Agricultural and Animal Husbandry College of Tibet University, Nyingchi, Linzhi 86000, 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. Chengdu Grain Storage Research Institute, State Administration of Grain Reserves, Chengdu 610031, 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