An efficient multiple shoot induction and genetic fidelity assessment of Exacum bicolor Roxb., an endemic and endangered medicinal plant
An efficient multiple shoot induction and genetic fidelity assessment of Exacum bicolor Roxb., an endemic and endangered medicinal plant
复制标题
DOI：
10.1007/s11627-015-9726-5
复制
发表时间：
2015-11
期刊：
影响因子：
--
通讯作者：
A. Ashwini;H. Ramakrishnaiah;S. H. Manohar;M. Majumdar
中科院分区：
文献类型：
--
作者：
A. Ashwini;H. Ramakrishnaiah;S. H. Manohar;M. Majumdar
双色黄药子（Exacum bicolor）为龙胆科（Gentianaceae）特有濒危药用植物。一个快速的协议已经开发了有效的多芽诱导测试节外植体上的Murashige和Skoog（MS）培养基补充各种细胞分裂素。单独使用细胞分裂素6-苄基腺嘌呤（BA）、6-糠基氨基嘌呤（Kn）、2-异戊烯基腺嘌呤（2-iP）和玉米素（Zn），并以不同浓度（0.5、1.0、2.0、5.0和10.0 μM）组合使用。结果表明，在10.0 μM BA + 2.0 μM Kn培养基中，不定芽数（19.33 ± 1.09个/外植体）、鲜重（5.1 ± 0.68 g）和干重（216.83 ± 2.84 mg）最高; 4周后，将来自琼脂培养物的多个芽继代培养到含有相同生长调节剂组合的液体培养基中。液体培养8 wk后，最佳处理的芽数增加了10倍左右（199.5 ± 1.14个/外植体）。以MS +3%蔗糖、10.0 μM BA + 2.0 μM Kn的全培养基为最佳，鲜重（13.76 ± 0.14 g）和干重（909.33 ± 1.92 mg）最高。添加0.5 μM吲哚-3-丁酸（IBA）30 d后，根系发育最快。将再生的植物成功地转移到含有椰子泥炭：泥炭混合物的盆中，并显示出75%的存活率。利用RAPD和ISSR标记对试管苗的遗传保真度进行了研究。所有的再生植株在遗传上都与它们的母株相同，在再生植株中没有显示出可检测到的遗传变异。因此，该方法可成功地用于大规模繁殖和种质保存。双色的
Exacum bicolor, an endemic and endangered medicinal plant, belongs to the family Gentianaceae. A rapid protocol has been developed for efficient multiple shoot induction by testing nodal explants on Murashige and Skoog (MS) medium supplemented with various cytokinins. The cytokinins 6-benzyladenine (BA), 6-furfurylaminopurine (Kn), 2-isopentenyladenine (2-iP) and zeatin (Zn) were used individually and in combination at different concentrations (0.5, 1.0, 2.0, 5.0, and 10.0 μM). The maximum number of shoots (19.33 ± 1.09 per explant) as well as their fresh weight (5.1 ± 0.68 g) and dry weight (216.83 ± 2.84 mg) were obtained with 10.0 μM BA + 2.0 μM Kn. After 4 wk, the multiple shoots from agar culture were subcultured into liquid medium containing the same growth regulator combinations. After 8 wk of liquid culture, the best treatment had about tenfold increase in shoot number (199.5 ± 1.14 per explant). The fresh weight (13.76 ± 0.14 g) and dry weight (909.33 ± 1.92 mg) were highest with full strength MS medium supplemented with 3% sucrose and containing 10.0 μM BA + 2.0 μM Kn. Maximum root development was observed after 30 d with 0.5 μM indole-3-butyric acid (IBA) supplementation. Regenerated plants were successfully transferred to pots containing coco peat:perlite mixture and showed a 75% survival rate. Genetic fidelity ofin vitroplantlets compared to mother plant were assessed using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR) markers. All the regenerated plants were genetically identical to their mother plant, showing no detectable genetic variation in the regenerated plantlets. Thus, this protocol could be successfully used for mass multiplication and germplasm conservation ofE. bicolor.