The present study reports enhanced in vitro mass propagation, synthetic seed production and its regeneration, acclimatization, and genetic fidelity assessment of in vitro regenerants of Bacopa monnieri (L.) Pennell; a medicinal plant renowned for its memory-enhancing property. For the first time, meta-Topolin (mT) as well as N⁶-benzyladenine (BA) and kinetin at their variable concentrations (0.5, 1. 1.5, 2 and 2.5 mg/l) were supplemented individually in Murashige and Skoog (Physiol Plant 15:473–497, 1962) (MS) basal medium to induce multiple shoots from shoot tip explants, initially collected from two-month-old vegetatively propagated plants. The best result in terms of multiplication (~ 7 shoots/explant) and biomass accumulation (~ 451.3 mg fresh weight, ~ 104.3 mg dry weight) was obtained in MS medium supplemented with 1 mg/l mT. For rooting of in vitro shoots, supplementation of 1.5 mg/l IAA and 0.5 mg/l IBA in MS media resulted in induction of the maximum number (~ 20) and highest length (~ 5.5 cm) of roots, respectively. Most efficient indirect regeneration and biomass production from green and hard basal calli (induced spontaneously in BA-supplemented media) was recorded in a simple MS medium. Synthetic seeds were produced from in vitro shoot tips using 2.5% sodium alginate and 75 mM calcium chloride solutions and were successfully regenerated in half-strength MS medium. Six-week-old in vitro regenerants were transferred to different substrates for acclimatization, among which cocopeat was found to be the best exhibiting the highest survival and growth in external conditions. Finally, molecular marker-(ISSR) and phenology-based genetic fidelity assessment of the in vitro regenerants exhibited that the in vitro regenerants were true-to-type in nature within themselves as well as with the mother plant.

本研究报告了Bacopa monnieri (L.) Pennell体外再生子的体外大规模繁殖、合成种子生产及其再生、驯化和遗传保真度评估；一种以增强记忆力而闻名的药用植物。第一次，meta -Topolin ( m T) 以及 N ⁶在 Murashige 和 Skoog (Physiol Plant 15:473–497, 1962) (MS) 基础培养基中分别补充不同浓度（0.5、1.1.5、2 和 2.5 mg/l）的 -苄基腺嘌呤 (BA) 和激动素，以诱导来自芽尖外植体的多个芽，最初是从两个月大的无性繁殖植物中收集的。在添加 1 mg/l m 的MS 培养基中获得了繁殖（~ 7 个芽/外植体）和生物量积累（~ 451.3 mg 鲜重，~ 104.3 mg 干重）方面的最佳结果T. 对于体外芽的生根，在 MS 培养基中补充 1.5 mg/l IAA 和 0.5 mg/l IBA 分别导致最大根数 (~ 20) 和最高长度 (~ 5.5 cm) 的诱导。在简单的 MS 培养基中记录了从绿色和硬基愈伤组织（在 BA 补充培养基中自发诱导）的最有效的间接再生和生物质生产。使用 2.5% 海藻酸钠和 75 mM 氯化钙溶液从体外芽尖产生合成种子，并在半强度 MS 培养基中成功再生。将 6 周龄的体外再生体转移到不同的基质中进行驯化，其中发现椰糠是最好的，在外部条件下表现出最高的存活率和生长率。最后，