【目的】探讨马兜铃经组织培养再生植株的条件.【方法】以马兜铃叶片作为外植体,以1/2MS、MS和改良MS培养基为基本培养基,在这些基本培养基上分别添加相应的植物激素而成为诱导马兜铃愈伤组织的培养基,观察:①3种基本培养基对诱导马兜铃愈伤组织的优劣;②不同pH值环境对诱导马兜铃愈伤组织的影响;③在相同的基本培养基中分别或同时加入不同种和不同量的植物激素(包括细胞分裂素KT或6-BA和生长激素NAA)后,对诱导马兜铃愈伤组织的影响。【结果】①在3种基本培养基中加入相同的植物激素(0.1mg/L的KT和0.2mg/L的NAA)后,MS和改良MS培养基均能诱导大量的愈伤组织,而1/2MS培养基则未能诱导;②相同的培养基(改良MS培养基加入0.1mg/L的KT和0.2mg/L的NAA)分别置于pH=5.8、pH=7.0、pH=8.0的环境中时,后两者均能诱导大量的愈伤组织,而前者则只能诱导很少量的愈伤组织;③在相同的基本培养基(改良MS培养基)中分别单独加入细胞分裂素和单独加入生长激素,前者未见诱导出愈伤组织,而后者能诱导出来;将不同种和不同量的植物激素组成各种组合加入到相同的基本培养基(改良MS培养基)中,所形成的各种培养基对诱导马兜铃愈伤组织的影响观察显示,加入生长激素过多时,细胞分裂过快,导致愈伤组织松散而不利于芽的分化,加入过少时愈伤组织诱导率不高;最佳的培养基组合是在改良MS培养基中加入1mg/LKT、0.5mg/L NNA的培养基。【结论】以MS或改良MS为基本培养基,并在此基础上添加1mg/L的KT和0.5mg/L的NAA组成诱导愈伤组织的培养基,在pH=7.0~8.0的碱性环境下培养,是马兜铃经组织培养能较好地诱导愈伤组织的条件。
[ Objective] To explore the culture conditions of inducing callus from tissue of Aristolochia contona Bge (ACB). [Methods] The leaves of ACB were used as the explants. Basic medium (including 1/2 MS medium, MS medium and modified MS medium) containing corresponding phyto-hormones was applied for the induction of ACB callus. The influences of different culture conditions such as three kinds of basic medium, different pH values and addition of different kinds of phyto-hormones at different concentrations into the basic medium respectively or simultaneously, on the callus induction of ACB were observed. [ Results] ( 1 ) After the three kinds of basic medium were added with the phytohormones of 0.1mg/L kinetin (KT) and 0.2mg/L s-naphthalene acetic acid (NAA), a large amount of ACB calluses were induced in the MS medium and modified MS medium, while ACB calluses did not occur in the 1/2 MS medium. (2) When the pH value of the culture medium composed of modified MS medium and 0. 1mg/L KT and 0.2mg/L NAA was at 5.8, 7.0 and 8.0, a large amount of ACB calluses were induced in the medium with pH value being 7.0 and 8.0 while a few calluses occurred in the medium with pH value being 5.8. (3) ACB calluses were induced in modified MS medium with NAA added , but calluses did not occur in modified MS medium with KT added. When the modified MS medium was added with different kinds of phyto-hormones at different concentrations, ACB calluses were loose in the medium with high-concentration NAA and this did not benefit to the differentiation of buds for too fast cell division, and the callus induction rate was low in the medium with low-concentration NAA. The optimized cuhure medium was modified MS medium with 1 mg/L KT and 0.5mg/L NAA added. [ Conclusion] The optimum culture conditions of inducing callus from ACB tissues are: MS medium or modified MS medium with lmg/L KT and 0.5mg/L NAA added beir,g the culture medium, and pH value being 7.0 - 8.0.