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如何在高等植物细胞中起作用？为了回答这个问题，我们从培养基中提取B后，观察了烟草BY-2培养细胞的形态学变化。用双醋酸荧光素（FDA）和碘化丙啶（P1）对细胞进行染色，以区分死细胞和活细胞。将烟草细胞转移到无b培养液中，细胞在9小时后开始死亡，36小时时细胞死亡率超过80%。在死亡细胞中，细胞质从细胞壁上分离出来，就像细胞被质解一样，P1均匀地染色细胞。另一方面，用30 mM H202或0.2% Triton X-100杀死细胞时，细胞核被P1强烈染色，而原生质的其他部分未被染色。从培养液中去除Ca离子也以与B类似的方式杀死细胞。为了使DNA转录可视化，我们将差异显示法应用于b缺陷细胞。在去除B后的3和6小时，在当前的实验条件下没有检测到任何显著的和可重复的变化。在治疗后18小时，检测到DNA转录的一些变化。所表达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.