Artemia Vulgaris Case Study

Plant material Seeds of Artemisia vulgaris L. were surface disinfected with 10% (v/v) dettol solution (Reckitt Benckiser, Kolkata, India) for 5 min, followed by rinsing three to five times in sterile distilled water. The seeds were then surface sterilized with 0.1% (w/v) aqueous mercuric chloride (HgCl2) for 4-5 min and finally rinsed with autoclaved distilled water (three to five changes). Surface sterilized seeds were inoculated into culture flasks containing 25 ml MS medium supplemented with 10% (v/v) filter sterilized coconut water and incubated in a dark chamber for 5-7 days at a temperature of 23C to facilitate germination. Later they were transferred to photoperiodic conditions and maintained for another 28-32 days for seedling growth.

vulgaris. Both the basal media were supplemented with 3% (w/v) sucrose and were solidified with 0.7% (w/v) agar and different concentrations of cytokinins including 0.44-13.32 M BA (benzyladenine), 0.46-13.92 M KN (kinetin) or 0.49-14.76 M 2iP (2-ispopentenyladenine).

2.4. Effect of carbon sources Shoot tip segments were cultured on MS basal medium supplemented with 4.92 M 2iP and different types of carbon sources, including 3% (w/v) of glucose, fructose and sucrose.

2.5. Effect of cytokinins Shoot tip segments were cultured on MS medium containing 3% (w/v) sucrose, 0.7% (w/v) agar and supplemented with different concentrations and combinations of cytokinins including 4.92 M 2iP + 0.444-13.32 M BA; 4.92 M 2iP + 0.464-13.92 M KN; 2.22 M BA + (0.464-13.92 M) KN + 4.92 M 2iP.

2.6. Effect of amino acids Shoot tip segments were excised and cultured on MS medium containing 30 g l-1 sucrose, 7.0 g l-1 agar supplemented with 4.92 M 2iP and 2.22 M BA and different concentrations of amino acids (glycine, Asparagine and proline ) (0.5 – 3.0 mg/l) individually and examined for improvement of shoot multiplication. The shoot sprouting frequency (%), number of shoots per explant and shoot length (cm) were recorded after 30 days of culture.

2.7.

Ten to twenty explants were used per treatment in each replication. Observations were recorded on the frequency (number of cultures responding for multiple shoot proliferation and root development) and the number of shoots per explant, shoot length, number of roots per shoot, root length, increase in plant height after successive hardening, number of inflorescence heads per plant, number of floral buds per inflorescence respectively. The analysis of variance (ANOVA) appropriate for the design was carried out to detect the significance of differences among the treatment means and the treatment means were compared using Duncan’s Multiple Range Test (DMRT) at a 5% probability level (Gomez and Gomez,