Somaclonal variation, often manifested as the increased ploidy of plants observed following in vitro culture, can be advantageous in ornamental species or those used for secondary metabolite production. Polyploidy occurs especially when plantlets are produced by protoplast and callus cultures. Plants were regenerated from green leaf mesophyll protoplasts of diploid Gentiana decumbens L.f. through somatic embryogenesis. A yield of more than 9 × 105 protoplasts per gram of fresh weight was achieved by incubating fully expanded young leaves in an enzyme mixture containing 1.0% (w/v) cellulase and 0.5% (w/v) macerozyme. Protoplasts, cultured in agarose beads using a modified Murashige and Skoog medium, divided and formed microcalli, with the highest plating efficiency obtained on medium containing 2.0 mg l-1 1-naphthaleneacetic acid and 0.1 mg l-1 thidiazuron. Callus proliferation was also promoted by including thidiazuron in agar-solidified medium, while somatic embryogenesis was induced from microcalli on medium supplemented with 1.0 mg l-1 kinetin, 0.5 mg l-1 gibberellic acid, and 80 mg l-1 adenine sulfate. Flow cytometric analysis and chromosome counting revealed that all regenerants were tetraploid.

中文翻译：

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从二倍体龙胆草Lf的叶肉原生质体间接体细胞胚发生再生自四倍体植物。

体细胞无性变异通常表现为在体外培养后观察到的植物倍性增加，在观赏性物种或用于次生代谢产物的物种中可能是有利的。多倍体特别是当通过原生质体和愈伤组织培养产生小植株时发生。通过体细胞胚发生从二倍体龙胆草Lf的绿叶叶肉原生质体再生植物。通过在含有1.0％（w / v）纤维素酶和0.5％（w / v）宏酶的酶混合物中孵育完全膨胀的幼叶，可以实现每克鲜重超过9×105原生质体的产量。使用改良的Murashige和Skoog培养基在琼脂糖珠中培养的原生质体分裂并形成微愈伤组织，在含有2.0 mg 1-1-1-萘乙酸和0的培养基上可获得最高的铺板效率。1毫克l-1噻地隆。通过在琼脂固化的培养基中加入噻唑隆也可以促进愈伤组织的增殖，而在补充有1.0 mg l-1激动素，0.5 mg l-1赤霉素和80 mg l-1腺嘌呤的培养基上，从微愈伤组织诱导体细胞胚发生。流式细胞仪分析和染色体计数显示所有再生体均为四倍体。