中药白芍愈伤组织的培养及其中芍药苷的测定

[目的]建立中药白芍愈伤组织培养的方法,并测定其中芍药苷的含量。[方法]以白芍叶、茎为外植体,接种于不同的愈伤诱导培养基和继代培养基上培养愈伤组织,并提取愈伤组织中的芍药苷,用TLC和HPLC法测定愈伤组织中芍药苷的含量。[结果]适合白芍茎的诱导愈伤培养基是1/2MS+0.2 mg/L 2,4-D+2.0 mg/L 6-BA+0.2 mg/L NAA,叶的最适诱导培养基是1/2MS+1.0 mg/L 2,4-D+1.5 mg/L 6-BA,白芍愈伤继代培养适合培养基为1/2MS+0.1 mg/L 2,4-D+1.0 mg/L ZT+0.5 mg/L NAA。经HPLC测定每克干白芍愈伤组织中的芍药苷含量为0.068 9 mg。[结论]中药白芍的叶和茎可诱导形成愈伤组织,且形成的愈伤组织中含有芍药苷。

Study on Callus Culture of Paeonia lactiflora pall and Determination of Paeoniflorin in Callus

[Objective] To establish the methods of callus culture,and determine the content of paeoniflorin in callus cells of Paeonia lactiflora pall.[Method] Leaves and stem sterilized were cut into tissue pieces and inoculated on different induction medium and regeneration medium.Paeoniflorin was extracted from callus and the content of paeoniflorin was determined by TLC and HPLC.[Result] The results showed that medium of 1/2MS＋0.2 mg/L 2,4-D＋ 2.0 mg/L 6-BA＋0.2 mg/L NAA were fit for stem callus induction,and medium of 1/2MS＋1.0 mg/L 2,4-D＋1.5 mg/L 6-BA were fit for leaves callus induction.Regeneration medium of callus is 1/2MS＋0.1 mg/L 2,4-D＋1.0 mg/L ZT＋0.5 mg/L NAA.The content of paeoniflorin of callus is 0.068 9 mg/g by HPLC.[Conclusion] Leaf and stem of Paeonia lactiflora pall could be induced to form callus.There are paeoniflorin in callus cells of Paeonia lactiflora pall.