K.A. Timiryazev Institute of Plant Physiology RAS, Botanicheskaya str., 35, Moscow, 127276, Russia.
Protoplasma. 2019 Jul;256(4):1153-1164. doi: 10.1007/s00709-019-01374-2. Epub 2019 Apr 9.
In this study, a simple and rapid DAPI-based protocol was developed and optimized to visualize polyphosphates (polyPs) in the cyanobacterium Synechocystis sp. PCC 6803. The optimum dye concentration and incubation time were determined, and formaldehyde fixation was shown to significantly improve polyP detection in Synechocystis cells. Using the developed protocol, for the first time, it was shown that 80% of Synechocystis cells under phosphate overplus were able to accumulate phosphorus as polyP 3 min after the addition of KHPO. After 1 h, the number of cells with polyP began to decrease, and after 24 h, polyP granules were detected in only 30% of the cells. Thus, the Synechocystis cells appeared to be heterogeneous in their ability to accumulate and mobilize polyP. Like other photosynthetic organisms, Synechocystis synthesized less polyP in the dark than in the light. The accumulation of polyP was not inhibited under conditions of cold and heat stresses, and some cells were even able to synthesize polyP at a temperature of approximately 0 °C.
在这项研究中,开发并优化了一种基于 DAPI 的简单快速方案,以可视化蓝藻集胞藻 PCC 6803 中的多聚磷酸盐(polyPs)。确定了最佳的染料浓度和孵育时间,并表明甲醛固定可显著提高集胞藻细胞中 polyP 的检测效果。使用所开发的方案,首次表明在磷酸盐过剩的情况下,80%的集胞藻细胞在添加 KHPO 后 3 分钟即可积累 polyP。1 小时后,开始有 polyP 的细胞数量减少,24 小时后,只有 30%的细胞中检测到 polyP 颗粒。因此,集胞藻细胞在积累和动员 polyP 的能力上似乎存在异质性。与其他光合生物一样,集胞藻在黑暗中的 polyP 合成量低于光照下。在冷、热应激条件下,polyP 的积累并未受到抑制,甚至有些细胞能够在接近 0°C 的温度下合成 polyP。