Synthesis of nonembryonic synseed, short term conservation, phytochemical evaluation and assessment of genetic stability through SCoT markers in Decalepis salicifolia

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2024-03-15 DOI:10.1007/s11240-024-02719-3

Adla Wasi, Anwar Shahzad, Sabaha Tahseen

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Abstract

The current work specifies an effort to conserve the highly endangered plant Decalepis salicifolia by utilizing encapsulation technology for high plantlet recovery, short-term storage, and conservation. In order to determine the best culture conditions for maximizing the ability of synseeds to develop into complete plantlets, nodal segments (NS) were encapsulated in a sodium alginate (SA) matrix. The best solution for creating isodiametric beads was determined to be a 3% sodium alginate with 100 mM CaCl₂ solution. Regeneration of the encapsulated segment was reported directly or after storage at 4 °C for up to 8 weeks. The highest shoot regrowth frequency (90.8%) and highest microshoots per encapsulated nodal segment i.e. 4.90 were recorded when Murashige and Skoog (MS) basal medium was supplemented with 5.0 µM 6-benzyl adenine (BA) + 1.0 µM indole-3-butyric acid (IBA) and encapsulated nodal segment were inoculated onto the nutrient medium comprised of MS + 5.0 µM BA. Microshoots rooted effectively on MS medium augmented with indole-3-butyric acid IBA (2.5 µM). Plantlets achieved from preserved synthetic seeds were acclimatized and relocated in the natural condition successfully with an immortality rate of 87.1%. Subsequently, it was planted in garden soil and exhibited no morphological changes. Gas Chromatography and Mass Spectrometry (GC-MS) of leaf extract, obtained from the donor plant as well as in vitro derived encapsulated regenerated plantlets shows the presence of diverse chemical compounds of immense pharmacological properties and number of biochemical compounds are almost similar in both, which established the biochemical similarity between them. Genetic similarity between the donar plant and the synseed-derived plant was confirmed by the presence of monomorphic bands produced with the help of the start codon targeted (SCoT) marker.

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通过 SCoT 标记合成盐肤木非胚胎合子、进行短期保存、植物化学评估和遗传稳定性评估

摘要当前的研究工作旨在利用封装技术实现小植株的高回收率、短期储存和保护，从而保护高度濒危的植物水黄皮（Decalepis salicifolia）。为了确定最佳培养条件，最大限度地提高合子发育成完整小植株的能力，将节片（NS）封装在海藻酸钠（SA）基质中。经确定，制造等径珠子的最佳溶液是 3% 的海藻酸钠和 100 mM CaCl2 溶液。据报道，封装的部分可直接再生，或在 4 °C 下储存长达 8 周后再生。当在 Murashige 和 Skoog（MS）基础培养基中添加 5.0 µM 6-苄基腺嘌呤（BA）+ 1.0 µM 吲哚-3-丁酸（IBA），并将封装的节段接种到由 MS + 5.0 µM BA 组成的营养培养基上时，记录到最高的芽再生频率（90.8%）和每个封装节段的最高微芽数，即 4.90 个。小芽在添加了吲哚-3-丁酸 IBA（2.5 µM）的 MS 培养基上有效生根。用保存的合成种子培育出的小植株在自然条件下成功地适应和迁移，不朽率达 87.1%。随后，它被种植在花园土壤中，形态没有发生变化。从供体植物和体外衍生的封装再生小植株中提取的叶提取物的气相色谱和质谱分析（GC-MS）显示，二者存在多种药理特性的化合物，而且生化化合物的数量几乎相似，这证实了二者在生化方面的相似性。在起始密码子定向（SCoT）标记的帮助下，单形带的存在证实了供体植物和同源植物之间的遗传相似性。

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