低温处理烟草子房利于对二胞原胚的离体操作

Low temperature treatment of biological material is a routine method of preservation, but the mechanism for this is still largely unknown. Here the influence of low temperature treatment of living ovary on two-celled proembryo operation in vitro was explored in tobacco. The influence was mainly positive. First, the efficiency of isolation of two-celled proembryo was significantly improved by 23.47% for one isolation experiment after treatment of the ovary at 4 C for 5-24 h. Cellular characteristics were then investigated in detail and the results showed that the cell viability and polarity of two-celled proembryos including organelle distribution, growth style and cell wall component distribution, were all well maintained after cold treatment. Furthermore, in vitro cultivation of two-celled proembryos was carried out and showed that they could still divide normally, the division process paralleling to that of two-celled proembryos isolated from fresh ovary. Unexpectedly, the division frequency of two-celled proembryos was significantly improved by cold treatment. These results demonstrate that low temperature treatment of tobacco ovary is of benefit for two-celled proembryo operation in vitro, by not only improving isolation efficiency, but also triggering the cell's division potential for further cultivation.