Plant regeneration from leaf-derived protoplasts within the Daucus genus: effect of different conditions in alginate embedding and phytosulfokine application

• DOI: 10.1007/s11240-014-0436-1
• Publication Date: 2014-02-04T02:48:19Z
• Authors: Katarzyna Maćkowska; Agata Jarosz; Ewa Grzebelus
• Keywords: Daucus carota; Protoplast

Protoplasts of four Daucus carota subspecies and three wild species were isolated from 2-week-old shoot cultures during overnight incubation in an enzyme mixture composed 1 % (w/v) cellulase Onozuka R-10 0.1 pectolyase Y-23. Before the culture, they embedded autoclave- or filter-sterilized alginate solution. Modified thin layer (TAL) extra film (ETAF) techniques applied for protoplast immobilization. A rich mineral-organic medium based on formulation Kao Michayluk supplemented with mg l−1 2,4-dichlorophenoxyacetic acid, 0.2 zeatin, optionally 100 nM phytosulfokine (PSK), a peptidyl plant growth factor, was used culture. Plating efficiency genotype-dependent 40-day-old cultures, it varied 10 pusillus to 73 D. subsp. sativus. considerably higher ability colony formation observed modified TAL culture system using presence PSK medium. Plant regeneration through somatic embryogenesis stimulated by application occurred five out seven accessions present study. We believe our data may facilitate use hybridization cultivated carrot.