Bergenia ligulata (Wall.): micropropagation, genetic fidelity, and SEM studies

IF 2.2 3区生物学 Q4 CELL BIOLOGY

In Vitro Cellular & Developmental Biology - Plant Pub Date : 2023-12-29 DOI:10.1007/s11627-023-10398-6

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引用次数: 0

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.

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

摘要 Bergenia ligulata，俗称 "Pashanbheda "或印度大黄，是一种多年生草本植物，因其多种多样的药用价值而广受认可。乱用 B. ligulata 使该物种濒临灭绝。这项研究旨在建立一个强大的组织培养方案，用于快速微繁殖 B. ligulata。该方案对于确保这一珍贵植物物种的可持续生产和防止其自然种群的减少至关重要。这项研究利用叶片和叶柄外植体，成功证明了叶状枝属植物体外再生的高效性。叶片或叶柄外植体上长出最多嫩枝的最有效组合是使用添加了 0.9 µM 和 1.8 µM 6-苄基氨基嘌呤（BAP）以及 0.5 µM 1-萘乙酸（NAA）的 Murashige and Skoog（MS）培养基。此外，在添加了 8.8 µM BAP 和 2.3 µM 激肽（Kn）的 MS 培养基上也能产生多芽。为了达到最佳生根效果，我们小心地分离出 45 天大的嫩枝，并将其置于半强度的 MS 培养基中。利用简单序列间重复（ISSR）进行的基于 PCR 的分子分析证实了再生小植株的基因克隆性质。约 80% 发育良好的体外再生植株在玻璃温室中适应了环境，从而表明了所开发方案的稳健性。在本研究的基础上，我们利用一种可重复的体外技术，在 1 年时间内实现了从单个外植体直接再生约 3597 株植株。这种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。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

In Vitro Cellular & Developmental Biology - Plant 生物-发育生物学

CiteScore

5.00

自引率

7.70%

发文量

审稿时长

6-12 weeks

期刊介绍： Founded in 1965, In Vitro Cellular & Developmental Biology - Plant is the only journal devoted solely to worldwide coverage of in vitro biology in plants. Its high-caliber original research and reviews make it required reading for anyone who needs comprehensive coverage of the latest developments and state-of-the-art research in plant cell and tissue culture and biotechnology from around the world.