Lin Li-Chang, Nalawade Satish Manohar, Mulabagal Vanisree, Yeh Mau-Shing, Tsay Hsin-Sheng
Department of Agronomy, National Chung Hsing University, Taichung, Taiwan.
Biol Pharm Bull. 2003 Oct;26(10):1467-71. doi: 10.1248/bpb.26.1467.
An efficient and rapid protocol for in vitro induction and complete plant regeneration of Polygonum multiflorum THUNB has been developed. Nodal explants were grown in vitro on Murashige and Skoog's (MS) basal medium containing different concentrations of alpha-naphthaleneacetic acid (NAA) and benzyladenine (BA). The nodal explants (97%) produced multiple shoots (4.7 shoots per explant) on MS basal medium supplemented with 0.2 mg/l NAA and 2.0 mg/l BA after 6 weeks of culture. Eighty-eight percent to 100% of the shoots (1.0 cm in length) elongated (about 3.02-4.28 cm) and rooted on MS basal medium supplemented with NAA or indole-3-butyric acid (IBA). All the rooted shoots were transferred to pots containing autoclaved soil, vermiculite, and peat moss (1 : 1 : 1). The plantlets were successfully acclimatized under greenhouse conditions with high humidity before transferring to the field. The anthraquinone contents were determined using HPLC. Analysis revealed that the contents of the major medicinal compounds-emodin and physcion in the 6 weeks old in vitro grown shoots and three month old in vitro propagated plants grown in greenhouse were higher than those of the marketed crude drug (processed underground or stem parts of P. multiflorum).
已开发出一种高效快速的何首乌离体诱导及植株完全再生方案。节段外植体在含有不同浓度α-萘乙酸（NAA）和苄基腺嘌呤（BA）的Murashige和Skoog（MS）基本培养基上进行离体培养。培养6周后，节段外植体（97%）在添加0.2 mg/l NAA和2.0 mg/l BA的MS基本培养基上产生多个芽（每个外植体4.7个芽）。88%至100%的芽（长度为1.0 cm）在添加NAA或吲哚-3-丁酸（IBA）的MS基本培养基上伸长（约3.02 - 4.28 cm）并生根。所有生根的芽被转移到装有经高压灭菌的土壤、蛭石和泥炭藓（1:1:1）的花盆中。在转移到田间之前，这些小植株在高湿度的温室条件下成功驯化。使用高效液相色谱法测定蒽醌含量。分析表明，6周龄的离体生长芽和温室中3个月龄的离体繁殖植株中主要药用化合物大黄素和大黄素甲醚的含量高于市售粗药（何首乌的地下加工部分或茎部）。
