Plant regeneration through somatic embryogenesis from anther cultures of C X R cultivar of coffee
Plant regeneration through embryogenesis has been achieved from cultured anthers of C X R cultivar of coffee (an interspecific hybrid derivative of Coffea congensis and C. canephora) (Robusta). Initially, flower buds were pretreated at different temperatures (5-25°C) for 24-72 h in dark. The anthers of uninucleate microspore stage were cultured on MS medium supplemented with different concentrations of auxins, cytokinins and L-cysteine HCI. Anthers excised from the floral buds were pretreated at 5°Cfor 48 h followed by 10°C for another 24 h and cultured on MS medium with IBA and BAP (benzyladenine) (1 mg/l each) for 90 days. After one more subculture on the same medium, they developed 1-2 mm wide, brown calli. After subculturing for another 90 dayson MS medium with 2,4-D (0.05 mg/l), IAA (0.5 mg/l) and kinetin (8.6 mg/l), the calli grew further to 3-4 mm thickness. When these calli were transferred subsequently onto half-strength MS medium supplemented with NAA (0.05 mg/l), kinetin (0.5 mg/l) and 2X KNO3, nodular, friable embryogenic calli were produced after two months from the original brown callus. The embryogenic calli in turn differentiated into 3-4 white somatic embryos in another month. These embryogenic calli and embryos were further multiplied on MS medium with ABA (1 mg/l). The embryos were converted into plantlets on half-strength MS medium with 0.1 mg/l of kinetin. Subsequently, these plantlets were hardened in net-pots and polybags.
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| Format: | biblioteca |
| Published: |
2000
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| Subjects: | COFFEA, COFFEA CANEPHORA, COFFEA CONGENSIS, CULTIVO DE ANTERAS, CAFE, HIBRIDOS, REGENERACION, EMBRIOGENESIS SOMATICA, ABA, AUXINAS, CITOQUININAS, AIA, AIB, HIBRIDACION INTERESPECIFICA, QUINETINA, CISTEINA, ANA, |
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