Callus formation from cotyledon protoplasts of Pinus oocarpa and Pinus patula |
| |
Authors: | E Lainé H David A David |
| |
Institution: | Lab. de Biologie et Physiologie végétales, Univ. de Bordeaux 1 –Avenue des Faculiés, F-33405, Talence Cedex, France. |
| |
Abstract: | Protoplasts were isolated from cotyledons of 11-day-old seedlings of Pinus oocarpa and P. patula ssp. tecunumanii . The best enzyme combination was Cellulase R10 + Pectolyase Y-23, associated with bovine serum albumin. When cultured at a low density 1.25 × 103 to 5 × 103 protoplast (ml)?1] in a liquid medium, the cells divided. The medium contained glutamine and casein hydrolysate as nitrogen sources, and glucose as osmoticum. Rate of division was increased by supplementing the medium with l -ornithine, putrescine and spermidine. However, the rate remained low, with an absolute division frequency of ca 1%. Dilution allowed colony proliferation and fragmentation, leading to the formation of numerous microcalli that could be transferred to various solid media for further growth. |
| |
Keywords: | Callus formation Pinus caribaea Pinus oocarpa Pinus patula polyamines protoplast density tropical pines |
|
|