Protoplast Isolation and Culture in Sugar Beet (Beta vulgaris L.)

In this study, attempts were made to optimize conditions for protoplast isolation from suspension and mesophyll cells, growth of isolated protoplasts and formation of callus from the cultured protoplasts. The enzyme mixture containing 1.0% hemicellulase, 10% pectinase and 0.1% driselase was found to be the most suitable combination for yield and viability of both suspension and mesophyll-drived protoplasts from sugar beet. Genotypic variation for plating efficiency was significant; ELK345 and M1017 lines producing significantly higher plating efficiency (PE) than other lines. On the other hand, suspension-derived protoplasts of line ELK345 formed more colonies than mesophyll-derived protoplasts of the same line. Micro-callus was obtained from only suspension-derived protoplasts of line CBM315. The effects of densities on PE of protoplast cultures were significant-i.e., the highest initial plating density (3 x 105 protoplasts ml-1) produced the highest PE. The differences between the levels of BAP or 2,4-D on colony formation of protoplasts were not significant. However, media containing combinations of BAP or 2,4-D were more efficient for cell colony formation than the control (i.e., hormone-free) medium.