A novel approach for direct shoot regeneration, anatomical characterization, and withanolides content in micropropagated plants of Withania somnifera (L.) Dunal—an important medicinal plant
{"title":"A novel approach for direct shoot regeneration, anatomical characterization, and withanolides content in micropropagated plants of Withania somnifera (L.) Dunal—an important medicinal plant","authors":"Ganesan Mahendran, Laiq ur Rahman","doi":"10.1007/s11627-024-10428-x","DOIUrl":null,"url":null,"abstract":"<p><i>Withania somnifera</i> (Ashwagandha) is a valuable therapeutic plant used in many Ayurvedic drugs marketed for health restoration. However, its commercial production faces challenges due to poor seed viability, low germination rates, and the absence of standardized propagation methods. Therefore, this investigation aimed to establish a fast and highly efficient method for direct shoot regeneration in <i>W. somnifera</i> by using the apical meristem perforation technique, which is a novel approach. Initially, rapid seed germination was standardized by soaking the seeds in distilled water for varying durations (0 to 48 h). The highest rates of shoot initiation (90.40 ± 0.51%) and mean number of direct shoot regenerations (16.20 ± 0.50 per explant) were achieved on MS basal medium fortified with meta-topolin (mT) at 3.0 mg L<sup>−1</sup>. For optimal root development, shoots (3 to 4 cm) were cultured on half-strength Murashige and Skoog (MS) medium with 3.0 mg L<sup>−1</sup> indole-3-butyric acid (IBA). Subsequently, well-rooted seedlings (90% survival rate) were acclimatized under glasshouse conditions in pots filled with a mixture of sandy loam and vermicompost (4:1). Anatomical examination of leaves and stems from micropropagated and acclimatized glasshouse plants revealed well-developed cuticles, differentiated mesophylls, and vascular bundles. Furthermore, the accumulations of withaferin A (1.26 ± 0.12 mg g<sup>−1</sup>), withanone (1.39 ± 0.18 mg g<sup>−1</sup>), and withanoside V (0.11 ± 0.04 mg g<sup>−1</sup>) in greenhouse-acclimated plant leaves were 1.88, 2.28, and 2.30 times higher, respectively, than in tissue culture samples (0.52 ± 0.07 mg g<sup>−1</sup>, 0.73 ± 0.02 mg g<sup>−1</sup>, and 0.05 ± 0.02 mg g<sup>−1</sup>). In conclusion, the described procedure offered an effective and straightforward method for large-scale production of <i>W. somnifera</i> plants for commercial metabolites.</p>","PeriodicalId":13293,"journal":{"name":"In Vitro Cellular & Developmental Biology - Plant","volume":"41 1","pages":""},"PeriodicalIF":2.2000,"publicationDate":"2024-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"In Vitro Cellular & Developmental Biology - Plant","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1007/s11627-024-10428-x","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"CELL BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Withania somnifera (Ashwagandha) is a valuable therapeutic plant used in many Ayurvedic drugs marketed for health restoration. However, its commercial production faces challenges due to poor seed viability, low germination rates, and the absence of standardized propagation methods. Therefore, this investigation aimed to establish a fast and highly efficient method for direct shoot regeneration in W. somnifera by using the apical meristem perforation technique, which is a novel approach. Initially, rapid seed germination was standardized by soaking the seeds in distilled water for varying durations (0 to 48 h). The highest rates of shoot initiation (90.40 ± 0.51%) and mean number of direct shoot regenerations (16.20 ± 0.50 per explant) were achieved on MS basal medium fortified with meta-topolin (mT) at 3.0 mg L−1. For optimal root development, shoots (3 to 4 cm) were cultured on half-strength Murashige and Skoog (MS) medium with 3.0 mg L−1 indole-3-butyric acid (IBA). Subsequently, well-rooted seedlings (90% survival rate) were acclimatized under glasshouse conditions in pots filled with a mixture of sandy loam and vermicompost (4:1). Anatomical examination of leaves and stems from micropropagated and acclimatized glasshouse plants revealed well-developed cuticles, differentiated mesophylls, and vascular bundles. Furthermore, the accumulations of withaferin A (1.26 ± 0.12 mg g−1), withanone (1.39 ± 0.18 mg g−1), and withanoside V (0.11 ± 0.04 mg g−1) in greenhouse-acclimated plant leaves were 1.88, 2.28, and 2.30 times higher, respectively, than in tissue culture samples (0.52 ± 0.07 mg g−1, 0.73 ± 0.02 mg g−1, and 0.05 ± 0.02 mg g−1). In conclusion, the described procedure offered an effective and straightforward method for large-scale production of W. somnifera plants for commercial metabolites.
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.