ISHS Acta Horticulturae 725: V International Symposium on In Vitro Culture and Horticultural Breeding
PROTOPLAST CULTURE IN PETUNIA HYBRIDA AND EFFECTS OF CELL CYCLE INHIBITORS ON CELL DIVISION FREQUENCY |
| Authors: | L. Meyer, M. Serek, T. Winkelmann |
| Keywords: | iodoacetamide, isolation, plating efficiency, regeneration, Rhodamine 6G |
| Abstract:
Protoplasts were isolated from six different Petunia hybrida genotypes and cultured embedded in alginate in liquid medium 8pm7 (1.0 mg L-1 benzylaminopurin (BAP) + 1.0 mg L-1 indole acetic acid (IAA) + 2.5% sucrose). This medium was replaced after seven to nine days by callus induction medium KF1 (0.5 mg L-1 BAP + 0.5 mg L-1 IAA and 1.8% sucrose) and later by KF2 (0.5 mg L-1 BAP + 0.5 mg L-1 IAA and 1.3% sucrose). Thereafter, calluses were transferred to solidified PET 2 medium (1 mg L-1 BAP + 2 mg L-1 IAA). The protoplast yield varied between 0.5 and 5.6 * 106 protoplasts per gram fresh weight. Cell division frequencies between 15 and 55% were observed. Shoot regeneration was obtained from three genotypes. In terms of a later fusion and selection of the heterologous hybrids the effects of the cell cycle inhibitors Rhodamine 6G (R6G) and Iodoacetamide (IOA) were analysed. The treatment with different concentrations of R6G and IOA reduced cell division markedly, but pronounced differences between experiments did not allow the identification of a appropriate concentration yet. | |
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