National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692 Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
Effective surface sterilization and shoot regeneration protocols for Clinacanthus nutans are described. Nodal segments of C. nutans were sterilised by single- and double-stage sterilisation methods. In the double-stage method, explants were soaked in Plant Preservative Mixture (PPM) at various concentrations (0, 1.25, 2.50, 5.0 µL/mL). For direct shoots regeneration, MS media were supplemented with TDZ, BAP, or KN in single or in combination (BAP+NAA or TDZ+NAA). For indirect shoot regeneration, single application (BAP or KN) and combination treatment (TDZ+BAP or TDZ+KN) of cytokinins were evaluated. The use of PPM (1.25 µL/mL) resulted in survival rate of explants at 82 ± 4.5 %. Direct shoot multiplication was the highest on the medium with 1.0 mg/L BAP + 0.1 mg/L TDZ where explants produced 20 ± 1.5 % primary, 57 ± 6.2 % secondary and 77 ± 6.6 % tertiary shoots respectively. The medium supplemented with BAP at 0.5 mg/L induced the highest number of shoots indirectly (3.5 ± 0.1 shoots/explant) while BAP at 1.5 mg/L induced the highest percentage of explant forming callus (84.4 ± 5.1 %). Therefore, double sterilisation method using Clorox and PPM at 1.25 µl/ml was optimum for surface sterilisation. A protocol for establishment of C. nutans culture is established.
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