Calcium and calcium ionophore A23187 induce high-frequency somatic embryogenesis in cultured tissues of <Emphasis Type="Italic">Coffea canephora</Emphasis> P ex Fr |
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Authors: | Akula Ramakrishna Parvatam Giridhar Gokare Aswathanarayana Ravishankar |
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Institution: | (1) Plant Cell Biotechnology Department, Central Food Technological Research Institute, Constituent Laboratory of Council of Scientific and Industrial Research, Mysore, 570 020, India; |
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Abstract: | High-frequency somatic embryogenesis was achieved in Coffea canephora using calcium ionophore A23187, which influences the influx of calcium into a cell. With 100 μM calcium ionophore and 5 mM
calcium, 85% and 70% of cultures produced embryogenic tissue, with 105 ± 7 and 95 ± 8 primary embryos from each callus mass
respectively. Medium supplemented with 100 μM EGTA (calcium chelator) or 1 mM verapamil (calcium channel blocker) significantly
reduced somatic embryogenesis. Calcium imaging studies were done to determine the relationship between morphogenetic response
and the cellular calcium levels. The calcium ionophore/calcium treatment was very effective in driving cellular machinery
toward embryogenesis. The embryos were regenerated into plantlets when cultured on MS medium supplemented with 5 mM calcium/100 μM
calcium ionophore A23187. Somatic embryogenesis-derived plants were successfully transferred to soil and grown to maturity
in the field. |
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