Efficient and rapid system of plant regeneration via protoplast cultures of Fagopyrum esculentum Moench

In the present study, a high yield of isolated protoplasts from the agronomically important crop Fagopyrum esculentum was obtained by applying a mixture of cellulase, pectolyase, and driselase. We demonstrated that the yield of morphogenic callus-derived protoplasts was 1 x 10(6) protoplasts per g of fresh tissue. For hypocotyls used as the protoplast source, the number of released cells was twice lower. The protoplasts, embedded in an agarose matrix and cultured in a modified Kao and Michayluk media supplemented with phytosulfokine, re-enter the cell cycle and start to develop, forming microcalli. The plating efficiency was about 20% in the case of hypocotyl- and morphogenic callus-derived protoplasts. For plant regeneration, the medium was supplemented with different combinations of cytokinin. Somatic embryogenesis and organogenesis occur during the cultivation of the protoplast-derived tissues, depending on the applied protoplast source. For the first time, an effective protoplast-to-plant system for F. esculentum has been developed. Key messageMorphogenic callus- and hypocotyl-derived protoplasts of buckwheat after embedding in agarose beads and culture in phytosulfokine enriched medium regenerated into plants.

Automated summary

By applying a mixture of cellulase, pectolyase, and driselase, a high yield of isolated protoplasts from buckwheat was obtained. Morphogenic callus-derived protoplasts showed a higher yield compared to hypocotyl-derived protoplasts. The protoplasts, embedded in an agarose matrix and cultured in a modified medium, successfully underwent cell cycle re-entry and development, leading to plant regeneration.