丹参微体快繁技术研究

Study on Micropropagation Technology of Salvia Miltiorrhiza

为建立丹参快速繁殖体系,本研究以丹参带节茎段为外植体,通过设置不同的消毒时间,研究用0.1%升汞溶液浸泡的消毒时间对丹参茎段污染率和褐化率的影响,在培养基中加入植物生长调节剂萘乙酸(NAA)、6-苄氨基腺嘌呤(6-BA),研究不同浓度的植物生长激素的配比对丹参诱导过程、扩繁过程和生根过程的影响,选择出最适宜的培养基配方,为丹参的组织培养提供技术支持。研究发现,丹参茎段使用75%乙醇消毒30 s,0.1%HgCl_(2)消毒10 min的效果最佳。腋芽诱导过程中,在添加1.5 mg/L 6-BA和0.1 mg/L NAA的培养基中,诱导率最高,此时芽体长势好,芽体壮为绿色,叶大,筛选出最适合丹参茎段的初代培养基的配方是MS+1.5 mg/L 6-BA+0.1 mg/L NAA。继代培养中,在MS培养基中加入1.0 mg/L 6-BA和0.4 mg/L NAA时增殖系数最高,增殖系数为12.83,丛生芽多且茎粗壮。生根过程中,在1/2 MS培养基中加入0.2 mg/L NAA时,生根率、平均生根数和平均根长都是四个处理中最高的,此时根系粗,根系多,有次生根。本研究筛选出的最佳初代、继代和生根培养基配方,为建立丹参离体快速繁殖程序和生产次生代谢物提供了基础支持。

In order to establish a rapid propagation system of Salvia miltiorrhiza,this study took Salvia miltiorrhiza as experimental material.The influence of the disinfection time soaked in 0.1% mercury solution on the contamination rate and Browning rate of stem segments was studied.Plant growth regulators naphthoacetic acid(NAA) and 6-benzyl amino adenine(6-BA) were added into the medium to study the effects of different concentrations of plant growth hormone on the induction process,propagation process and rooting process of Salvia miltiorrhiza.The most suitable medium formula was selected to provide technical support for the tissue culture of Salvia miltiorrhiza.The results showed that the stem segment of Salvia miltiorrhiza was suitable to be sterilized with 75% ethanol for 30 s and 0.1%HgCl_(2) for 10 min.In the initial culture,the induction rate of axillary buds was the highest in the medium supplemented with 1.5 mg/L 6-BA and 0.1 mg/L NAA.At this time,the buds grew well,the buds were strong and green with large leaves.The optimum medium for stem segment of Salvia miltiorrhiza was MS+1.5 mg/L 6-BA+0.1 mg/L NAA.In the subculture,when 1.0 mg/L 6-BA and 0.4 mg/L NAA were added into MS medium,the proliferation coefficient was the highest(12.83),with many tufted shoots and stout stems.In the rooting process,when 0.2mg/L NAA was added into 1/2 MS medium,the rooting rate,average number of roots and average root length were the highest among the four treatments.In this case,the roots were thick,with multiple roots and secondary roots.In this study,the optimal formula of primary,secondary and rooting medium was selected,which pointed out the development direction for solving the problems faced by artificial cultivation of Salvia miltiorrhiza,establishing rapid propagation program in vitro and producing secondary metabolites.