Bioscience and Biotechnology Center, Nagoya University, Nagoya, Japan.
Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Nagoya, Japan.
J Plant Res. 2022 Sep;135(5):693-701. doi: 10.1007/s10265-022-01406-8. Epub 2022 Jul 14.
Plasmodesmata are unique channel structures in plants that link the fluid cytoplasm between adjacent cells. Plants have evolved these microchannels to allow trafficking of nutritious substances as well as regulatory factors for intercellular communication. However, tracking the behavior of plasmodesmata in real time is difficult because they are located inside tissues. Hence, a system was constructed to monitor the movement of substances by plasmodesmata using tobacco BY-2 cells, which are linearly organized cells, and a microfluidic device that traps them in place and facilitates observation. After targeting one cell for photobleaching, recovery of the lost H2B-GFP protein was detected within 200 min. No recovery was detected in that time frame by photobleaching the entire cell filaments. This suggested that the recovery of H2B-GFP protein was not due to de novo protein synthesis, but rather to translocation from neighboring cells. The transport of H2B-GFP protein was not observed when sodium chloride, a compound known to cause plasmodesmata closure, was present in the microfluid channel. Thus, using the microfluidic device and BY-2 cells, it was confirmed that the behavior of plasmodesmata could be observed in real time under controllable conditions.
胞间连丝是植物中连接相邻细胞之间细胞质的独特通道结构。植物进化出这些微通道，以允许营养物质以及细胞间通讯的调节因子的运输。然而，由于它们位于组织内部，因此实时跟踪胞间连丝的行为是困难的。因此，构建了一个使用烟草 BY-2 细胞和微流控装置的系统，该系统可以监测胞间连丝的物质运动，烟草 BY-2 细胞是线性排列的细胞，而微流控装置可以将它们固定在适当的位置并便于观察。在用光漂白靶向一个细胞后，在 200 分钟内检测到丢失的 H2B-GFP 蛋白的恢复。在该时间范围内用光漂白整个细胞丝，没有检测到恢复。这表明 H2B-GFP 蛋白的恢复不是由于从头合成蛋白质，而是由于从相邻细胞的易位。当微流道中存在氯化钠（一种已知会导致胞间连丝关闭的化合物）时，没有观察到 H2B-GFP 蛋白的运输。因此，使用微流控装置和 BY-2 细胞，证实可以在可控条件下实时观察胞间连丝的行为。
