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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. [ABSTRACT FROM AUTHOR]

Bergenialigulata, 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. [ABSTRACT FROM AUTHOR]

An efficient in vitro propagation and synthetic seed production protocol was established for the conservation of Decalepis salicifolia (Bedd. ex Hook.f.) Venter, an endemic and critically endangered ethnomedicinal species. Murashige and Skoog (MS) media supplemented with 2.2 μM BAP and 5.7 μM IAA produced shoot tip proliferation with an average shoot length of 5.81 ± 0.03 cm in 3 wk. Axillary bud proliferation was found to be best in 11.1 μM BAP with an average of 3.5 ± 0.06 shoots per explant. Shoot tip and nodal explants were encapsulated in sodium alginate and their regeneration was achieved following storage at 4°C up to 12 wk. Successful rooting was obtained in modified MS medium with low nitrate and high sucrose concentration. Quarter strength MS media containing 2.5 mM each of NH4NO3 and KNO3 along with 8% sucrose produced an average number of 12.4 ± 1.18 roots with an average length of 4.3 ± 0.08 cm. The in vitro derived rooted plants were successfully hardened (92.8%) and established in field with 100.0% survival rate. Inter-simple sequence repeat (ISSR) markers proved the genetic fidelity among the in vitro grown plant showing 99.5% similarity with the mother plant. Micropropagation derived acclimatized plants produced 2-hydroxy-4-methoxybenzaldehyde in amounts similar to seed-derived field-grown plants of the same age. The in vitro propagation protocol developed for D. salicifolia can be utilized to reduce exploitation pressure from their natural habitats and augment ecorestoration, conservation, and cultivation of this critically endangered and industrially valuable plant. The synthetic seeds technique will serve as propagules in ex situ gene banks, as well as supports cost-effective germplasm conservation. [ABSTRACT FROM AUTHOR]