In vitro multiplication of Coffea arabica L. from leaf explants through indirect somatic embryogenesis
Esayas Aga, Yashwant Khillare
Leaf explants of approximately one year old C. arabica L. plant samples from India were surface sterilized and cultured on MS (Murashige and Skoog) medium containing 30g/l sucrose, 4g/l agar and 10mg/l ascorbic acid. Out of the different MS media supplemented with different combination of auxins and cytokinins tested in a separate experiment for callus induction from the leaf explants, only six auxin/cytokinin combinations induced callus formation. Callus induction occurred in the range of three to four weeks after the inoculation of the explants. Each callus initiated on specific culture medium was sub cultured on identical but fresh medium at an interval of four weeks after callus initiation. After two months of incubation in the callus induction medium, the calli were transferred on to half strength MS medium of their respective callus induction medium for the initiation of somatic embryos and plantlets. After four weeks of incubation on these half strength MS media somatic embryos developed on the surface of the callus as green color. In the subsequent additional sub-culturing/month for four months on the same half strength MS media, the developed somatic embryos matured and germinated into plantlets (shoots). The 10 cm shoots were rooted on and acclimatized.The results were discussed from identification of the best growth hormone combinations and their respective concentrations suitable for Coffea Arabica L. in vitro multiplication from leaf explants through indirect somatic embryogenesis.