A regenerative protocol and SEM study for in vitro propagation of Anthurium crossed lines via indirect somatic embryogenesis

Abstract

A reproducible protocol for indirect somatic embryogenesis was established in the crosses of Anthurium. Two new lines namely-A1P and A2W were formed from the crosses of Anthurium ornatum and A. andraeanum. Leaf explants were selected based on coloured spathe having fragrances spadix (pink-A1P and white-A2W). Half-strength modifi ed MS (lowering ammonium nitrate to 250 mg/l and for the fi rst time-0.1% EDTA Ferric Sodium) medium supplemented with 2, 4-D (1.4 μM) and 6-BAP (4.4 μM) was found to induce high percentage (82.6±0.57) of callus after 30 days in dark conditions (A2W). Later, MS basal medium having 6-BAP (0.2-4.4 μM) was found to initiate shoot proliferation from the calluses of both the lines. The highest number of adventitious shoots were obtained in A2W(29.1±0.88) compared to A1P (26.8±0.33), at 1.3 μM6-BAP after 6 weeks in 16/8 h light and dark cycle under a photoperiod of 50 μmol/m2/s. Separate rooting media was not required, and the best rooting occurred with 0.4 μM and 1.3 μM6-BAP shooting media. The results showed that A2W line had different responses compared to A1P line, both for callus induction as well as number of shoots-per-explants. Shoots for both the lines increased in the multiplication stages and this increase in the number of shoots-per-explants was found signifi cant (p<0.01). The scanning electron microscopic (SEM) study also confi rmed the stages of somatic embryogenesis from the leaf explants.