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An effective protocol was developed for in vitro propagation of Psoralea corylifolia via somatic embryogenesis in cell suspension culture. Embryogenic callus was obtained on Murashige and Skoog (MS) medium supplemented with 6 µM naphthaleneacetic acid (NAA) and 30 µM glutamine from transverse TCLs from 10-day-old hypocotyl explants with a 96.4% frequency. Embryogenic callus produced a higher number of somatic embryos (123.7 ± 1.24 per gram fresh weight callus) onMS medium containing 30 g l⁻¹ sucrose, 1 µM NAA, 4 µM benzyladenine (BA), 15 µM glutamine and 2 µM abscisic acid (ABA) after 4 weeks of culture. Somatic embryos successfully germinated (97.6%) on ½ MS medium containing 20 g l⁻¹ sucrose, 8 g l⁻¹ agar and supplemented with 2 µM BA, 1 µM ABA and 2 µM gibberellic acid (GA₃) within 2 weeks of culture. Somatic embryos developed into normal plants, which hardened with 100% efficiency in soil in a growth chamber. Plants were successfully transferred to greenhouse and subsequently established in the field. Plant survival percentage in the field differed with seasonal variations. Average psoralen content of 12.9 µg g⁻¹ DW was measured in different stages of somatic embryo development by high-performance liquid chromatography (HPLC). This protocol will be helpful for efficient propagation of elite clones on a mass scale, conservation efforts of this species and for secondary metabolites production studies.
This paper describes a protocol to develop cotton mutant lines with improved yield and fiber characters. Immature ovules [(15-day post-anthesis ovules (dap)] were irradiated with 10–50 Gy gamma rays and treated with 1–5 mM EMS and SA to investigate the stimulatory effects of mutagenic treatments. During the subsequent field trials, the mutant lines showed significant variations from control lines. Lower dose/concentration of mutagenic treatments effectively stimulate the agronomical characters like early flowering, plant height, number of bolls, yield of seed cotton, ginning percent, seed index, harvest index and fiber characters while exposure at higher dose/concentration results in lowering the value of the parameters. Consequently, we use this approach to induce genetic variability for obtaining novel mutant cotton cultivars. Among the 19 different mutant lines isolated from the study, M7 showed higher morphological variations in terms of yield characters such as plant height, number of bolls, yield of seed cotton, ginning percent and fiber characters. Significant increase in cellulose content was also noted in mutant lines, whereas moderate increase was observed in total fiber units of the mutant lines. The selected cotton mutant lines for cultivars were investigated systematically; these lines significantly increased the potential for agronomical enhancement of cotton yield.
A rapid and efficient micropropagation system was developed for Psoralea corylifolia, an endangered, valuable medicinal plant. Multiple shoot buds were obtained in half-strength liquid Phillips–Collins (L2) medium supplemented with 5 lM benzylaminopurine (BA) and 5 μM thidiazuron (TDZ) from apical bud explants of 1-week-old cultures. The shoot buds were subcultured on enriched solid L2 medium supplemented with different concentrations and combinations of BA, kinetin (KIN), 2-isopentenyladenine (2iP), TDZ, bavistin (BVN) and trimethoprim (TMP). Enriched solid L2 medium supplemented with 2 μM BA, 1 μM TDZ and 100 mg l⁻¹ BVN were more effective in producing greater number of shoots per explant (85.2 ± 0.9 shoots/explant) after 4 weeks of culture. The regenerated shoots (40– 50 mm in length) rooted and accompanied by hardening upon transfer to 50 μM indole-3-butyric acid (IBA) for 15 min and followed by planting in sterile soil mixture and vermiculate (3:1 v/v), with 50 ml of one-eight strength L2 basal salt solution devoid of sucrose and inositol, supplemented with 5 μM IBA and 100 mg l⁻¹ BVN. The plants achieved 100% rooting with hardening. Subsequently the rooted plants were successfully established in the field. The survival percentage differed with seasonal variations. The concentration of psoralen was evaluated in different tissues of ex vitro and in vivo grown plants by high-performance liquid chromatography (HPLC). Psoralen content was increased in leaves (2.97%), roots (2.38%), stems (5.40%) and seeds (1.63%) of ex vitro plants than the in vivo plants. This system facilitates for commercial and rapid propagation of P. corylifolia for conservation strategies and phytomedicine production.
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