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利用节间外植体通过直接器官发生进行微繁殖:扫描电子显微镜、气相色谱 - 质谱联用及起始密码子靶向(SCoT)标记分析

Micropropagation of via Direct Organogenesis Using Internodal Explants: SEM, GC-MS, and SCoT Marker Analysis.

作者信息

Kausar Arisha, Shahzad Anwar, Bhat Aashiq Yousuf, Ramakrishnan Muthusamy, Ahmad Zishan

机构信息

Plant Biotechnology Section, Department of Botany, Aligarh Muslim University, Aligarh 202002, India.

State Key Laboratory of Tree Genetics and Breeding, Co-Innovation Center for Sustainable Forestry in Southern China, Bamboo Research Institute, School of Life Sciences, Nanjing Forestry University, Nanjing 210037, China.

出版信息

Plants (Basel). 2025 Aug 12;14(16):2507. doi: 10.3390/plants14162507.

Abstract

is a herb with high medicinal value and a low range of distribution. It is used in several herbal and traditional medicines, including diabetes. In the present study, we designed the methodology for the micropropagation of from internodal segments. The highest shoot multiplication was achieved on Murashige and Skoog (MS) medium supplemented with 6-benzyl-amino-purine (BAP) (5.0 µM) + indole-3-acetic acid (IAA) (1.5 µM) + adenine sulphate (ADS) (15.0 µM), which produced the maximum number of 20.45 ± 0.12 shoots/explants with 6.43 ± 0.006 cm shoot length. Rooting in the microshoots was attained on half-strength MS medium containing indole-3-butyric acid (IBA) (1.5 µM), with the highest root number of 16.44 ± 0.015 roots/shoot, and root length of 2.25 ± 0.011 cm. To assess genetic fidelity, SCoT marker analysis was performed on nine randomly selected regenerated plantlets and the mother plant, all of which exhibited monomorphic banding patterns, confirming genetic stability. Scanning electron microscopy (SEM) reveals normal stomatal structure in the regenerated plants post-acclimatization, indicating successful physiological recovery. Furthermore, Gas Chromatography-Mass Spectrometry (GC-MS) analysis confirms the presence of major phytocompounds in both the regenerated plants and the mother plant, supporting the conservation of phytochemical integrity. Given the restricted distribution and overharvesting pressure on this species, the established protocol provides an efficient strategy for rapid, large-scale, and genetically stable propagation to support conservation and pharmaceutical utilization.

摘要

是一种具有高药用价值且分布范围狭窄的草本植物。它被用于多种草药和传统药物中,包括治疗糖尿病。在本研究中,我们设计了从节间段进行微繁殖的方法。在添加了6-苄基腺嘌呤(BAP)(5.0 μM)+吲哚-3-乙酸(IAA)(1.5 μM)+硫酸腺嘌呤(ADS)(15.0 μM)的Murashige和Skoog(MS)培养基上实现了最高的芽增殖,该培养基产生的最大芽数为20.45±0.12个芽/外植体,芽长为6.43±0.006厘米。在含有吲哚-3-丁酸(IBA)(1.5 μM)的1/2强度MS培养基上实现了微芽生根,最高根数为16.44±0.015条根/芽,根长为2.25±0.011厘米。为了评估遗传稳定性,对9个随机选择的再生植株和母株进行了SCoT标记分析,所有植株均表现出单态带型,证实了遗传稳定性。扫描电子显微镜(SEM)显示驯化后的再生植株气孔结构正常,表明生理恢复成功。此外,气相色谱-质谱(GC-MS)分析证实再生植株和母株中均存在主要的植物化合物,支持了植物化学完整性的保存。鉴于该物种分布受限且面临过度采收压力,所建立的方案为快速、大规模和遗传稳定的繁殖提供了一种有效策略,以支持保护和药用利用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5152/12389594/a4dbc1459e47/plants-14-02507-g001.jpg

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