Analysis of gene expression induced by boron deficiency.

高等植物需要硼(B)作为必需的营养元素。B是如何在高等植物细胞中发挥作用的？为了回答这一问题，观察了烟草BY-2培养细胞从培养液中去除B后的形态变化，并用二乙酸荧光素(FDA)和碘化丙啶(P1)对细胞进行染色，以区分死亡细胞和活细胞。当烟草细胞被转移到无B的培养液中时，细胞在9小时后开始死亡，36小时后80%以上的细胞死亡。在死亡细胞中，细胞质与细胞壁分离，就像细胞质壁被质壁溶解一样，细胞内的P1染色均匀。30 mM H_(202)或0.2%Triton X-100杀灭细胞后，胞核呈浓染，而胞质的另一部分不染。去除培养液中的钙离子也会以类似于B-DID的方式杀死细胞。为了直观显示DNA转录，差异显示方法被应用于B-缺陷细胞。在去除B后3小时和6小时，在当前实验条件下没有检测到任何显著和可重复性的变化。治疗后18h，可检测到DNA转录的几个变化。表达的DNA的克隆目前正在进行中。

Higher plants require boron (B) as an essential nutritional element. How does B work in higher plant cells? To answer this question, the morphological changes of the tobacco BY-2 cultured cells after withdrawal of B from the culture media were followed.The cells were stained with Fluorescein diacetate (FDA) and Propidium iodide (P1) to discern dead and alive cells. When the tobacco cells were transferred to B-free culture solutions, the cells started to die after 9 hours and at 36 hours more than 80% of the cell died. In dead cells, protoplasm was separated from cell walls as if the cells were plasmolysed and P1 stained evenly the cells. On the other hand, when the cells were killed with 30 mM H202 or 0.2% Triton X-100, the nuclei were stained intensely with P1 while the other part of the protoplasm was not stained. Removal of Ca ions from the culture solution also killed the cells in a similar manner as B did.To visualize the DNA transcription, the differential display method was applied to B-deficient cells. At 3 and 6 hours after the B removal, any significant and reproducible changes were not detected under the current experimental condition. At 1 8 hours after the treatment, several changes in the DNA transcription were detected. Cloning of the expressed DNA is now under progress.