Reshi Zubair Altaf, Husain Fohad Mabood, Khanam Mehrun Nisha, Javed Saad Bin
Plant Biotechnology Laboratory, Department of Botany, Aligarh Muslim University, Aligarh, 202002, India.
Department of Food Science and Nutrition, College of Food and Agriculture Sciences, King Saud University, Riyadh, KSA, Saudi Arabia.
BMC Plant Biol. 2025 Jan 28;25(1):121. doi: 10.1186/s12870-025-06095-8.
An efficient in vitro propagation protocol has been established for a valuable medicinal plant, Salix tetrasperma using mature nodal explants. The investigation aimed to observe the influence of various combinations and concentrations of cytokinins (mT, BA, and Kn) and auxins (NAA, IAA, and IBA) on regeneration potential using the Murashige and Skoog (MS) medium. Among individual cytokinin treatments, 5.0 µM mT resulted highest response of 92% with maximum shoot number (11.6 ± 0.08) per explant and shoot length (4.5 ± 0.22 cm) after 12 weeks of culture. However, synergistic treatment of mT (5.0 µM) and NAA (0.5 µM) further improved proliferation with (21.3 ± 0.40) shoots per explant and (6.9 ± 0.13 cm) shoot length in 96% cultures after 12 weeks of incubation. Rooting from in vitro raised microshoots was achieved on ½ MS medium supplemented with various concentrations of low-dose auxins. The highest number of roots (10.4 ± 0.20) per shoot with mean root length (5.7 ± 0.11 cm) with maximum rooting frequency (97%) was observed in 0.5 µM IBA, after 4 weeks of culture. The rooted plantlets achieved a remarkable 86% survivability rate, when transferred to ex vitro conditions during acclimatization. Analysis of photosynthetic parameters and their characteristics during the acclimatization phase revealed a gradual decline in photosynthetic attributes during initial weeks; however, a significant improvement was noted as the growth proceeded. SEM analysis revealed the ultra-morphological structural differences between in vivo and in vitro derived leaves of S. tetrasperma. Moreover, DPPH assay observed differential antioxidant activity of in vitro raised plantlets throughout the acclimatization period. The GC-MS analysis from leaf extracts of donor plants and in vitro derived plantlets has revealed a broad spectrum of phytochemical compounds with significant pharmacological properties. No polymorphism in the banding pattern was found when the genetic fidelity of the regenerated plants was evaluated using SCoT primers, indicating the clonal stability of micropropagated plants. This study is the first to explore the use of mT in regeneration of S. tetrasperma, showing its more effectiveness than BA and Kn.
利用成熟节段外植体，已为一种有价值的药用植物四籽柳建立了高效的离体繁殖体系。本研究旨在观察不同组合和浓度的细胞分裂素（mT、BA和Kn）和生长素（NAA、IAA和IBA）对使用Murashige和Skoog（MS）培养基再生能力的影响。在单独的细胞分裂素处理中，5.0 µM mT处理的响应率最高，为92%，培养12周后每个外植体的芽数最多（11.6±0.08），芽长最长（4.5±0.22厘米）。然而，mT（5.0 µM）和NAA（0.5 µM）的协同处理进一步提高了增殖效果，培养12周后，96%的培养物中每个外植体的芽数为（21.3±0.40），芽长为（6.9±0.13厘米）。在添加不同浓度低剂量生长素的1/2 MS培养基上，实现了离体培养的微芽生根。培养4周后，在0.5 µM IBA处理中观察到每个芽的根数量最多（10.4±0.20），平均根长为（5.7±0.11厘米），生根频率最高（97%）。在驯化过程中转移到体外条件下时，生根苗的成活率显著达到86%。对驯化阶段光合参数及其特征的分析表明，在最初几周光合属性逐渐下降；然而，随着生长的进行，有显著改善。扫描电镜分析揭示了四籽柳体内和体外叶片的超微形态结构差异。此外，DPPH分析观察到离体培养的植株在整个驯化期内具有不同的抗氧化活性。对供体植物和离体培养植株的叶片提取物进行的GC-MS分析揭示了具有显著药理特性的多种植物化学化合物。使用SCoT引物评估再生植株的遗传稳定性时，未发现条带模式的多态性，表明微繁殖植株的克隆稳定性。本研究首次探索了mT在四籽柳再生中的应用，表明其比BA和Kn更有效。
