Abstract: Akebia trifoliate (Thunb.) Koidz. has high development and application value, and it is very important to establish the tissue culture system for its excellent strains. Taking the young branches and leaves of Akebia trifoliate (Thunb.) Koidz. as explants, this paper studied the processes of aseptic explants, callus induction and embryoid induction, and analyzed the factors influencing the inhibition of callus browning. The results showed that: after 75% ethanol disinfection of 1 min and mercuric chloride sterilization of 12 min, aseptic explants were obtained. After soaking 3 min in sterile water, then explants were inoculated to the culture medium (MS + NAA 0.4 mg·L-1 +2, 4D 4.0 mg·L-1 + characoal activited 1.0 g·L-1 + sucrose 30.0 g·L-1 + agar 6.0 g·L-1) culturing without light, callus could be obtained, and the browning of callus could be effectively inhibited. The obtained callus inoculated on the media of MS + NAA 0.4 mg·L-1 +6BA 2.0 mg·L-1 + sucrose 30.0 g·L-1 + agar 6.0 g·L-1 could obtain the embryos, and then seedlings could be obtained after inoculation into a hormone free MS medium.

Key words: Akebia trifoliate (Thunb.) Koidz., callus induction, browning, differentiation

摘要： 三叶木通（Akebia trifoliate (Thunb.) Koidz.）具有很高的开发应用价值，其优良品系组织培养快繁体系的建立尤为重要。以三叶木通幼嫩枝条及叶片作为外植体，研究了三叶木通无菌外植体获得、愈伤诱导以及胚状体诱导等过程，同时分析了抑制愈伤组织褐化的影响因素。结果表明：外植体经75%乙醇处理1 min，0.1%升汞灭菌12 min，再经无菌水浸泡处理3 min后，接种至MS+NAA 0.4 mg·L-1+2,4D 4.0 mg·L-1+活性炭1.0 g·L-1+蔗糖30.0 g·L-1 +琼脂6.0 g·L-1培养基中，避光培养可得到愈伤组织，并能有效抑制愈伤组织褐化。愈伤组织接种至MS+NAA 0.4 mg·L-1+6BA 2.0 mg·L-1+蔗糖30.0 g·L-1 +琼脂6.0 g·L-1的培养基中可得到胚状体，然后接种至无激素的MS培养基中，可以得到种苗。

关键词: 三叶木通, 愈伤诱导, 褐化, 分化