Postgraduate and Research Department of Botany, St. Thomas College, Pala, Arunapuram, Kottayam (Dt.), Kerala, India.
Appl Biochem Biotechnol. 2013 Mar;169(6):1799-810. doi: 10.1007/s12010-013-0117-2. Epub 2013 Jan 23.
An efficient plant regeneration protocol was established for an endangered ethnomedicinal plant Desmodium gangeticum (Linn.) DC. Morphogenic calli were produced from 96 % of the cultures comprising the immature leaf explants on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (4.0 mg l(-1)) in combination with 6-benzylaminopurine (BA; 0.8 mg l(-1)). For callus regeneration, various concentrations of BA (1.0-5.0 mg l(-1)) or thidiazuron (TDZ; 1.0-5.0 mg l(-1)) alone or in combination with indole-3-acetic acid (IAA; 0.2-1.0 mg l(-1)) were used. Highest response of shoot regeneration was observed on MS medium fortified with TDZ (4.0 mg l(-1)) and IAA (0.5 mg l(-1)) combination. Here, 100 % cultures responded with an average number of 22.3 shoots per gram calli. Inclusion of indole-3-butyric acid in half MS medium favored rooting of recovered shoots. Out of 45 rooted plants transferred to soil, 40 survived. Total DNA was extracted from the leaves of the acclimatized plants of D. gangeticum. Analysis of random amplified polymorphic DNA using 13 arbitrary decanucleotide primers showed the genetic homogeneity in all the ten plants regenerated from callus with parental plant, suggesting that shoot regeneration from callus could be used for the true-to-type multiplication of this plant.
建立了一种高效的药用植物印度蔊菜（Desmodium gangeticum (Linn.) DC.）的植物再生方案。在添加了 2,4-二氯苯氧乙酸（4.0 mg·l^(-1)）和 6-苄基氨基嘌呤（BA；0.8 mg·l^(-1)）的 Murashige 和 Skoog (MS) 培养基上，由 96%的幼叶外植体组成的培养物中产生了形态发生愈伤组织。为了进行愈伤组织再生，单独或组合使用不同浓度的 BA（1.0-5.0 mg·l^(-1)）或噻二唑脲（TDZ；1.0-5.0 mg·l^(-1)）以及吲哚乙酸（IAA；0.2-1.0 mg·l^(-1)）。在添加 TDZ（4.0 mg·l^(-1)）和 IAA（0.5 mg·l^(-1)）的 MS 培养基上，观察到最高的芽再生反应。在此，100%的培养物每克愈伤组织平均产生 22.3 个芽。在半 MS 培养基中加入吲哚丁酸有利于恢复芽的生根。在转移到土壤中的 45 株生根植物中，有 40 株存活。从适应环境的 D. gangeticum 植物叶片中提取总 DNA。使用 13 个任意的十聚体引物对随机扩增多态性 DNA 的分析表明，所有 10 株从愈伤组织再生的植物与亲本植物具有遗传同质性，表明从愈伤组织再生的芽可用于该植物的真实类型繁殖。
