黄连的组培再生及遗传转化研究.pdfVIP

·2460 · 中草药 Chinese Traditional and Herbal Drugs 第 44 卷 第 17 期 2013 年 9 月 黄连的组培再生及遗传转化研究 吴平平，郭惠芳，蔺自敏，姚明镜* 华中农业大学植物科学技术学院药用植物研究所，湖北 武汉 430070 摘 要：目的 建立黄连的组培再生及遗传转化体系。方法 以黄连子叶和下胚轴为外植体，用不同的基本培养基添加不同 种类及浓度的植物生长调节剂进行组培再生研究；利用基因枪转化法研究黄连胚性愈伤组织和遗传转化。结果 子叶和下胚 轴在 MS 培养基上愈伤诱导率分别可达 86.31%和 54.34%；与MS 培养基相比 6, 7-V 培养基可获得更高的愈伤诱导率和增殖 率；诱导愈伤组织最适宜的激素组合为 0.5 mg/L 2, 4-D＋0.5 mg/L KT ；不同细胞分裂素培养的愈伤组织颜色和质地差异明显； 0.5 mg/L KT ＋0.5 mg/L IAA 与 1.0 mg/L 6-BA＋0.5 mg/L NAA 组合可诱导出胚性愈伤组织，产生体细胞胚；无激素的 6, 7-V 培养基可维持胚性愈伤组织的不断生长；含 1 mg/L GA ＋0.5 mg/L IBA 的 6, 7-V 培养基可使体细胞胚萌发成苗；转化后的 3 胚性愈伤组织经 3 mg/L 除草剂Basta 筛选，检测到 β-葡萄糖苷酸酶（GUS ）阳性，PCR 检测再生植株膦丝菌素乙酰转移酶 基因（bar ）阳性。结论 建立并完善了黄连的组织培养再生体系，并在此基础上利用转基因技术获得了具有相关性状的植 株，实现了黄连的遗传转化，为黄连性状的遗传改良奠定了基础。 关键词：黄连；愈伤组织；体细胞胚；组培再生；遗传转化 中图分类号：R282.21 文献标志码：A 文章编号：0253 - 2670(2013)17 - 2460 - 06 DOI: 10.7501/j.issn.0253-2670.2013.17.022 Tissue culture regeneration and genetic transformation of Coptis chinensis WU Ping-ping, GUO Hui-fang, LIN Zi-min, YAO Ming-jing Institute of Medicinal Plant, College of Plant Science and Technology of Huazhong Agricultural University, Wuhan 430070, China Abstract: Objective To establish the system of tissue culture regeneration and gene transformation for Coptis chinensis. Methods The cotyledon and hypocotyl of C. chinensis were used as explants, and the effects of different basic media with different plant growth regulators on in vitro tissue culture regeneration were compared; The embryogenic calli of C. chinensis were used as recipients for genetic transformation by particle bombardment method. Results The induction rates of cotyledon and hypocotyl were 86.31% and 54.34%, respectively; The 6, 7-V medium was more suitable for callus induction and proliferation rate of C. chinensis than the MS medium. The optimal hormone combination for callus