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 GA₃＋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。

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 induction was 0.5 mg/L 2,4-D + 0.5 mg/L KT in MS medium; Either the appropriate concentration of cytokinin alone or the combination of both cytokinin and auxin could induce the continuous proliferation of calli; However, only the hormone combination 0.5 mg/L KT + 0.5 mg/L IAA and 1.0 mg/L 6-BA + 0.5 mg/L NAA could generate the somatic embryos. 6, 7-V medium in absence of hormone could maintain the continuous growth of the embryogenic calli. The somatic embryos could germinate and grow up to plantlets on 6, 7-V medium containing 1 mg/L GA₃ + 0.5 mg/L IBA. The embryogenic calli were transformed by particle bombardment method and screened under 3 mg/L Basta, then the activity of β-glucuronidase was detected. PCR analysis showed that the bar gene was successfully transferred to the regenerated plants. Conclusion The tissue culture regeneration and genetic transformation system of C. chinensis is established, which lays the foundation for its genetic improvement.

教育部高校自主科研专项华中农业大学资助项目（52204-07021）