Effect of liquid pulses with 6-benzyladenine on the induction of somatic embryogenesis from coffee (Coffea arabica L.) callus cultures |
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Authors: | Iosif Papanastasiou Katerina Soukouli Georgia Moschopoulou Jane Kahia Spiridon Kintzios |
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Institution: | (1) Laboratory of Plant Physiology, Faculty of Agricultural Biotechnology, Agricultural University of Athens, Iera Odos 75, Athens, 11855, Greece;(2) International Coffee Research Institute, Ruiru, Kenya |
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Abstract: | We investigated the effect of the physical state of the nutrient medium on the induction of somatic embryogenesis on cell
cultures derived from coffee (Coffea arabica L.). Non-embryogenic callus tissues were pulsed initially with 50 μM 6-benzyladenine (BA) for 6, 24 or 48 h in half-strength
liquid Murashige and Skoog (MS) medium. After pretreatment, calli were transferred to agar-solidified half-strength MS medium
supplemented with 50 μM BA (‘standard induction medium’). Control callus tissues were incubated directly on the solid standard
induction medium. Callus growth was promoted by longer pretreatment periods. Formation of globular somatic embryos was observed
on callus tissues pretreated with BA for 24 or 48 h, which developed fully to cotyledonary-stage within only 2 weeks after
transfer to agar-solidified medium supplemented with BA. No embryo formation occurred in control cultures. Pretreatment with
BA in liquid medium was associated with changes in the redox status of cultured cells, such as alterations of the ascorbate–glutathione
redox systems and the accumulation of free radicals and oxidized lipids, as well as the possible reduction of cytochrome c-mediated apoptotic pathways. In particular, the induction of somatic embryogenesis was highly positively correlated (r
2 = 0.822) with the accumulation of protein carbonyls. The physiological role of BA as an inducer of both embryonic differentiation
and cellular death is discussed. |
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Keywords: | Cell culture Embryo germination Leaf explant Indirect somatic embryogenesis Redox status |
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