| 摘要 |
FIELD: agriculture.SUBSTANCE: invention relates to the field of plant biotechnology and forestry. The invention is a method of cryopreservation of axillary buds of aspen plants in vitro, comprising insulation of axillary buds, their preliminary dewatering in media containing osmolytes, transfer of the buds to the cryovials, cryopreservation of the cryovials with the buds in liquid nitrogen, thawing the buds and post-cryogenic regeneration from the of plants characterized in that on the step of dehydration before quick freezing the buds are placed first in a solution I containing nutrient medium and osmolytes (WPM supplemented with sucrose (0.2-0.5M), glycerol (1.7-2.5M)), then the buds are transferred into the solution II containing nutrient medium and osmolytes (WPM, sucrose (0.2-0.5M), glycerol (2.5-3.5M), ethylene glycol (1-1.5), dimethylsulfoxide (1.5-2M)), followed by transfer into the liquid for freezing (WPM, containing sucrose (0.2-0.5M), glycerol (3.5-4M), ethylene glycol (1.5-2.5M), dimethyl sulfoxide (1 5-2M)).EFFECT: invention enables to carry out the direct regeneration from the buds, providing the genetic safety in vitro of cultures of valuable selection genotypes and genetically modified aspen clones and survival to 80% after the cryo-storage for a year.2 cl, 3 dwg, 1 tbl |
