Alatar Abdulrahman A, Faisal Mohammad, Abdel-Salam Eslam M, Canto Tomas, Saquib Quaiser, Javed Saad B, El-Sheikh Mohamed A, Al-Khedhairy Abdulaziz A
Department of Botany & Microbiology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia.
Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain.
Saudi J Biol Sci. 2017 Sep;24(6):1430-1436. doi: 10.1016/j.sjbs.2017.03.008. Epub 2017 Mar 16.
In the present study, we develop an efficient and reproducible regeneration system for two cultivars , Jamila and Tomaland of L., an economically important vegetable crop throughout the world. Sterilization of seeds with 2.5% (v/v) NaOCl was found to be most effective, about 97% of seeds germinated on cotton in magenta box moistened with sterile half strength (½)Murashige and Skoog (MS) medium. Regeneration efficiency of cotyledonary leaf (CL) and cotyledonary node (CN) explants derived from 08 days old aseptic seedling were assessed on MS medium supplemented with different concentrations of auxins and cytokinin. CL explants were found more responsive in comparison to CN in both the cultivars. Types of basal media were also assessed and found to have a significant effect on shoot regeneration. Highest regeneration frequency and maximum number of shoots were standardized from CL explants on MS medium supplied with 6-benzyl adenine (BA; 5.0 µM), indole-3-butyric acid (IBA; 2.5 µM) and Kinetin (Kin; 10.0 µM). regenerated microshoots were rooted on ½MS medium containing 0.5 µM indole-3-butyric acid (IBA). Regenerated plantlets with well-developed roots and shoot system were successfully acclimated to condition. Genetic uniformity of tissue culture raised plantlets was first time evaluated using flow cytometry and single primer amplification reaction (SPAR) methods ., DAMD and ISSR. No significant changes in ploidy level and nuclear DNA content profile were observed between propagated plants and normal plants of both the cultivars. Similarly, the SPAR analysis also revealed monomorphic banding patterns in regenerated plantlets of verifying their genetic uniformity and clonal fidelity. This efficient regeneration system can be used as a fast and reproducible method for genetic transformation of this important vegetable crop.
在本研究中，我们为茄子（Solanum melongena L.）的两个品种Jamila和Tomaland开发了一种高效且可重复的再生系统，茄子是全球一种经济上重要的蔬菜作物。发现用2.5%（v/v）次氯酸钠对种子进行消毒最为有效，在装有湿润无菌半强度（½）Murashige和Skoog（MS）培养基的洋红色盒中的棉花上，约97%的种子发芽。在添加不同浓度生长素和细胞分裂素的MS培养基上评估了源自8日龄无菌幼苗的子叶（CL）和子叶节（CN）外植体的再生效率。在两个品种中，CL外植体均比CN外植体反应更灵敏。还评估了基础培养基类型，发现其对芽再生有显著影响。在添加6-苄基腺嘌呤（BA；5.0 μM）、吲哚-3-丁酸（IBA；2.5 μM）和激动素（Kin；10.0 μM）的MS培养基上，CL外植体的再生频率最高且芽的数量最多。再生的微芽在含有0.5 μM吲哚-3-丁酸（IBA）的½MS培养基上生根。根系和芽系统发育良好的再生植株成功适应了环境条件。首次使用流式细胞术和单引物扩增反应（SPAR）方法，即DAMD和ISSR，评估了组织培养再生植株的遗传一致性。两个品种的组培繁殖植株与正常植株之间未观察到倍性水平和核DNA含量谱的显著变化。同样，SPAR分析也显示再生植株中条带模式单一，证实了它们的遗传一致性和克隆保真度。这种高效的再生系统可作为这种重要蔬菜作物遗传转化的快速且可重复的方法。
