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A well-organized procedure was established for the conservation and distribution of Sphagneticola calendulacea (L.) Pruski [synonym Wedelia chinensis (Osbeck) Merrill] for the first time, using alginate-encapsulated nodal segments (NSs) as synthetic seeds. The ideal beads were obtained through a combination of 2.5% sodium alginate and 75 mM calcium chloride with 84.40 ± 2.20% rate of shoot emergence. The maximum regeneration (88.84 ± 2.24%) from synthetic seeds was achieved on liquid 1/2Murashige and Skoog (MS) medium in comparison to its other formulations. Furthermore, superior frequency (91.09 ± 2.24%) of complete plantlet (having both shoots and roots) formation was achieved when synthetic seeds were cultured on liquid 1/2MS (1.5% sucrose) fortified with 1.0 mg L⁻¹ N₆-benzyladenine plus 0.25 mg L⁻¹ α-naphthalene acetic acid. Synthetic seeds could be effectively stored at low temperature (8 °C) up to 90 days with a survival rate of 52.38 ± 3.06%, whereas higher temperature (25 °C) did not support regeneration after 75 days of storage. The plantlets were successfully acclimatized to natural conditions with ~ 89% survival frequency. To by-pass the time-consuming in vitro culture step after encapsulation, synthetic seeds were directly regrown into complete plantlets ex vitro on sand, soil, and vermicompost (1:1:1; w/w). Regeneration rate of 42.22 ± 2.22% was attained when NSs were pretreated on 1/2MS medium containing 4.0 mg L⁻¹ indole-3-acetic acid for 24 h in dark, prior to encapsulation. The random amplified polymorphic DNA and intersimple sequence repeat fingerprinting profiles demonstrated genetic uniformity amongst the regenerated plantlets, in vitro mother plant, as well as in vivo wild plant.
Plant regeneration by means of somatic embryogenesis has been standardized for the first time in Eclipta alba (L.) Hassk. Explants like nodal segment, shoot tip, and leaf were tested on Murashige and Skoog medium supplemented with picloram, 2,4-dichlorophenoxyacetic acid, or α-naphthalene acetic acid for callus induction. Nodal segment exhibited the maximum response (92.80%) to callus induction on culture medium-containing 2.0 mg l⁻¹ picloram. Furthermore, highest rate (88.40%) and number (34.83) of somatic embryo induction were found on medium fortified with 2.0 mg l⁻¹ picloram, 0.5 mg l⁻¹ thidiazuron, and 0.25 mg l⁻¹ abscisic acid. Influence of abscisic acid on the somatic embryo regeneration was also tested with the highest frequency (95.50%) attained on medium-containing 0.75 mg l⁻¹ N⁶-benzyladenine and 0.5 mg l⁻¹ abscisic acid. Somatic embryo of torpedo stage was efficiently encapsulated with 2.5% sodium alginate and 75 mM calcium chloride, and exhibited 93.33% germination rate. The study revealed that storage at low temperature (8 °C) gave superior results where 86.67% of the synthetic seeds sprouted even after 60 days with a decline to 46.67% in 90 days post-storage. The rate of germination at 25 °C had been constantly low and failed to sprout after 75 days. The acclimatization of the regenerated plantlets was successfully taken place in garden soil, sand, and vermicompost in the ratio of 1:1:1 with 95% survival rate.
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