Plant regeneration from protoplasts of Pastinaca sativa L. via somatic embryogenesis

Abstract

In the present study we report the development of an effective and relatively efficient protocol for protoplast-to-plant regeneration of parsnip (Pastinaca sativa L.) via indirect somatic embryogenesis. The regenerative potential of three open-pollinated and four hybrid cultivars was assessed. The protoplast isolation efficiency after digestion of source material in an enzyme mixture consisted of 1% cellulase Onozuka R-10 and 0.1% pectolyase Y-23 reached on average 3.6 × 106 of cells per g of fresh mass. Protoplasts embedded in an alginate matrix and cultured in parsnip protoplast culture medium with phytosulfokine-α and putrescine reconstructed their cell wall and re-entered mitotic divisions. After the release from alginate, microcallus proliferated continuously on Gamborg B5 medium with vitamins supplemented with 100 nM of phytosulfokine-α. Indirect somatic embryogenesis occurred during the callus culture of cultivar ‘Półdługi biały’. The regenerated and acclimatized plants were morphologically similar to their donors and displayed no variation in the ploidy level.