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目的:对三角叶黄连愈伤组织的培养条件进行优化,建立三角叶黄连组织培养的技术体系。方法:采用响应面方法(RSM)中的Box-Behnken设计方法优化三角叶黄连愈伤组织增殖的最佳激素配比;考察不同培养基、外植体、温度、光照条件对三角叶黄连愈伤组织诱导的影响。结果:以茎尖为外植体愈伤组织诱导率最高,为12.67%,最佳培养条件为(15±2)℃,暗培养。2,4-D,KT是影响三角叶黄连愈伤组织形成的主要因素,其次为NAA。结论:三角叶黄连愈伤组织诱导的最适培养基为6,7-V+2,4-D 0.5 mg.L-1+KT 2.0 mg.L-1,增殖的最适培养基为6,7-V+2,4-D 4.6 mg.L-1+KT 3.8 mg.L-1+NAA 2.4 mg.L-1。
OBJECTIVE: To optimize the culture conditions of Coptis chinensis callus and to establish the tissue culture system of Coptis chinensis. Methods: The best hormone ratio of Coptis chinensis callus proliferation was optimized by Box-Behnken design method in Response Surface Methodology (RSM). The effects of different media, explants, temperature and light on callus Tissue-induced effects. Results: The highest callus induction rate was 12.67%. The best culture condition was (15 ± 2) ℃, dark culture. 2,4-D, KT is the main factor affecting the formation of Coptis chinensis callus, followed by NAA. CONCLUSION: The optimal culture medium for the treatment of Coptis chinensis callus is 6,7-V + 2,4-D 0.5 mg.L-1 + KT 2.0 mg.L-1, the optimum medium for proliferation is 6, 7-V + 2,4-D 4.6 mg.L-1 + KT 3.8 mg.L-1 + NAA 2.4 mg.L-1.