Atropa acuminata Royle ex Lindl. (family Solanaceae) is a critically endangered therapeutic herb, known for its immense therapeutic potential against innumerable ailments. In view of protecting this species from brink of extinction, present work focuses upon development of an effectual micropropagation protocol via indirect organogenesis using root-derived callus. Best callusing was observed on Murashige and Skoog (MS) medium augmented with 10 µM Thidiazuron (TDZ), where 99% of root explants induced whitish-green and compact calluses within 45 days. Upon sub-culturing on same medium (10 µM TDZ), these callus pieces differentiated an average of 5.54 ± 0.65 shoots having 2.38 ± 0.37 cm of length in 71.5% of the explants, after 28 days. MS + 1 µM indole-3-butyric acid (IBA) resulted in optimum rooting response where an average of 8.08 ± 0.19 roots having an average length of 7.28 ± 0.11 cm were produced. Micropropagated plants with well-developed and thick roots were acclimatized under greenhouse conditions. Genetic fidelity of ex vitro acclimatized plants using start codon targeted (SCoT) and sequence-related amplified polymorphism (SRAP) markers, revealed 96% similarity between mother plant and micropropagated plants. These plants showed higher yield of phenols, flavonoids and exhibited better antioxidant (DPPH and phosphomolybdenum assay) activities in comparison with mother plant. Higher levels of photosynthetic pigments and antioxidant enzymes were also observed in these plants as compared to the mother plant. Besides, quantification of atropine and scopolamine using high-performance liquid chromatography (HPLC) in different plant parts of acclimatized plantlets revealed higher amount of both the alkaloids in leaves. The present protocol shall be gainfully employed for large-scale micropropagation, conservation and extraction of biotherapeutic molecules for commercial purposes.

Atropa acuminataRoyle前Lindl。（茄科）是一种极度濒临灭绝的草药，以其对多种疾病的巨大治疗潜力而闻名。为了保护该物种免于灭绝的边缘，目前的工作集中在通过使用根衍生的愈伤组织的间接器官发生发展有效的微繁殖方案。在添加10 µM噻二唑隆（TDZ）的Murashige和Skoog（MS）培养基上观察到最佳的愈伤组织，其中99％的根外植体在45天内诱导发白绿色和紧凑的愈伤组织。在相同培养基（10 µM TDZ）上进行亚培养后，经过28天，这些愈伤组织碎片在71.5％的外植体中平均分化出5.54±0.65枝长，长度为2.38±0.37 cm。MS + 1 µM吲哚-3-丁酸（IBA）导致最佳的生根响应，平均值为8.08±0。产生19个平均长度为7.28±0.11cm的根。在温室条件下使根部发达且根粗的微繁殖植物适应环境。使用起始密码子靶向（SCoT）和序列相关的扩增多态性（SRAP）标记对离体驯化植物的遗传保真度显示，母本植物与微繁植物之间有96％的相似性。与母本植物相比，这些植物显示出更高的酚类，黄酮类化合物产量，并表现出更好的抗氧化剂（DPPH和磷钼含量测定）活性。与母本植物相比，在这些植物中还观察到更高水平的光合色素和抗氧化酶。除了，使用高效液相色谱（HPLC）对适应的小苗的不同植物部位中阿托品和东pol碱的定量分析表明，叶片中两种生物碱的含量均较高。本协议应有益地用于商业用途的生物治疗分子的大规模微繁殖，保存和提取。