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本研究在正交设计的基础上,以短柱铁线莲的叶片、叶柄、茎段为外植体,进行愈伤组织培养,并探讨愈伤组织褐化的抑制。研究结果表明:短柱铁线莲诱导愈伤组织的适宜外植体为茎段,诱导率为89.8%;合适的愈伤诱导培养基为1/2MS+0.4 mg/L TDZ+0.2 mg/L NAA+10 g/L蔗糖,诱导率为90%;合适的增殖培养基为1/2MS+30 g/L蔗糖+0.2 g/L肌醇+0.2 g/L水解络蛋白(CH)+5 mg/L Vc+0.3 mg/L TDZ+0.2 mg/L NAA,增殖率为302.3%。适宜不定芽诱导培养基为1/2MS+30 g/L蔗糖+0.76%琼脂+0.2 g/L肌醇+0.2 g/L水解络蛋白(CH)+0.3 mg/L TDZ+0.1 mg/L NAA,诱导率达63.3%。培养基中添加5 mg/L Vc或1 g/L活性炭时,可以能够很好地抑制短柱铁线莲褐化。
In this study, based on orthogonal design, the leaves, petioles and stems of Clematis sibirica were used as explants for callus culture and the inhibition of callus browning was discussed. The results showed that the suitable explants for the induction of callus were the stem segments with the induction rate of 89.8%. The suitable callus induction medium was 1 / 2MS + 0.4 mg / L TDZ + 0.2 mg / L NAA + 10 g / L sucrose, the induction rate was 90%; suitable proliferation medium was 1/2 MS + 30 g / L sucrose + 0.2 g / L myo-inositol + 0.2 g / L hydrolyzate + / L Vc + 0.3 mg / L TDZ + 0.2 mg / L NAA, the proliferation rate was 302.3%. Adventitious bud induction medium was 1 / 2MS + 30 g / L sucrose + 0.76% agar + 0.2 g / L inositol + 0.2 g / L hydrolyzed protein (CH) + 0.3 mg / L TDZ + 0.1 mg / L NAA , The induction rate of 63.3%. When 5 mg / L Vc or 1 g / L activated carbon was added to the medium, the browning of the stichopus japonicus could be well inhibited.