Abstract

Bergenia ligulata, commonly known as ‘Pashanbheda’ or Indian rhubarb, is a perennial herb that has been recognized for its diverse medicinal properties. The indiscriminate use of B. ligulata has brought the species to the brink of becoming threatened. This research aims to establish a robust tissue culture protocol that can be utilized for the rapid micropropagation of B. ligulata. This protocol is essential for ensuring the sustainable production of this valuable plant species and preventing the depletion of its natural populations. The study successfully demonstrated an efficient in vitro regeneration in B. ligulata, using leaf and petiole explants. The most effective combination for achieving the highest number of shoots on either explant (leaf or petiole) involved using Murashige and Skoog (MS) medium supplemented with 0.9 µM and 1.8 µM 6-benzylamino purine (BAP) with 0.5 µM 1-naphthaleneacetic acid (NAA). Moreover, multiple shoots were also produced on MS medium fortified with 8.8 µM BAP and 2.3 µM kinetin (Kn). To achieve optimal rooting, the 45-d-old shoot was carefully isolated and placed in a half-strength MS medium. PCR-based molecular analysis using inter simple sequence repeats (ISSR) confirmed the genetically clonal nature of regenerated plantlets. About 80% of the well-developed in vitro regenerated plants were acclimatized in the glasshouse, thereby showing the robustness of the developed protocol. Based on the present study, a reproducible in vitro technique was utilized to achieve direct regeneration of approximately 3597 plants from a single explant over a 1-yr period. This approach involved molecular fidelity analysis and scanning electron microscopy (SEM) analyses to ensure reliable results.

中文翻译：

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Bergenia ligulata（墙）：微繁殖、遗传保真度和 SEM 研究

摘要

岩 白菜，俗称“Pashanbheda”或印度大黄，是一种多年生草本植物，因其多种药用特性而受到认可。滥用B. ligulata已使该物种濒临受到威胁。本研究旨在建立一个强大的组织培养方案，可用于舌状芽孢杆菌的快速微繁殖。该协议对于确保这种有价值的植物物种的可持续生产和防止其自然种群的枯竭至关重要。该研究成功地证明了使用叶和叶柄外植体在B. ligulata中进行有效的体外再生。要在外植体（叶或叶柄）上获得最高数量的芽，最有效的组合涉及使用 Murashige 和 Skoog (MS) 培养基，并补充有 0.9 µM 和 1.8 µM 6-苄氨基嘌呤 (BAP) 以及 0.5 µM 1-萘乙酸（ NAA）。此外，还在用 8.8 µM BAP 和 2.3 µM 激动素 (Kn) 强化的 MS 培养基上产生了多个芽。为了实现最佳生根，将 45 天的芽仔细分离并置于半强度 MS 培养基中。使用简单序列重复 (ISSR) 的基于 PCR 的分子分析证实了再生植株的遗传克隆性质。约 80% 发育良好的体外再生植物在温室中适应，从而显示了所开发方案的稳健性。根据本研究，利用可重复的体外技术在一年内从单个外植体中直接再生出约 3597 株植物。该方法涉及分子保真度分析和扫描电子显微镜 (SEM) 分析，以确保结果可靠。