These methods can deliver a continuous supply of plant materials from elite germplasm lines, which might help exploit the therapeutic properties of these plant species and eradicate the want for harvesting specimens in the wild. Thomas and Yoichiro standardized an in vitro propagation protocol for the rare medicinal plant Justicia gendarussa applying nodal explants, and this enhanced system for plant regeneration is useful for the study of phytochem ical production, Balaraju et al. reported an effective regeneration protocol to get a useful medicinal plant, Vitex agnus castus, and all regenerated plants exhibited higher homo geneity, Within a earlier study, tissue culture and plant regeneration through direct shoot organogenesis induced from the shoot tip or axially bud of H. pogonocalyx was reported, However, indirect shoot organogenesis from leaf explants has in no way been reported within this species.
In our previous study, the 95% ethanol extract of H. pogo nocalyx exhibited cost-free radical scavenging activities, Therefore, the objective of this study was to develop a easy and hugely effective regeneration protocol making use of leaf explants and selleckchem examine the antioxidant activities in the regenerated plants. The compounds from regener ated plants of H. pogonocalyx have been also isolated, and their structures and activities had been evaluated. Methods Plant material Hygrophila pogonocalyx Hayata was collected in the Highlands Experiment Farm, National Taiwan University, Taiwan and identified by Mr. Chi Luan Wen, Seed Improvement and Propagation Station, Council of Agriculture, Taiwan. A voucher specimen was deposited in the Graduate Institute of Pharmacognosy, Shoot proliferation and plant regeneration For shoot organogenesis, young leaves were used as ex plants and cultured on Murashige and Skoog basal medium supplemented with BA, NAA, IAA, or 2iP at various concentrations, as shown in Table 1.
The media were supplemented with 3% su crose and solidified with 0. 7% agar, along with the pH was adjusted to 5. 7. order Triciribine The adventitious buds rooted and regener ated into plantlets when cultured on MS medium without plant regulators. For the mass production of plantlet, six to eight node explants cut from a regenerated plantlet had been cultured in sterile vessels with 100 ml of liquid MS medium supplemented with 3% sucrose. The rooted plantlets have been transplanted to a potting mixture with garden soil. The potted plants were acclimatized for four weeks then transferred to the field. Extract preparation The aerial parts of plants have been harvested month-to-month, fro zen at 80 C for 24 h, and lyophilized for 48 h.