A simple method to produce Synechocystis PCC6803 biofilm under laboratory conditions for electron microscopic and functional studies
A simple method to produce Synechocystis PCC6803 biofilm under laboratory conditions for electron microscopic and functional studies
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一种在实验室条件下产生集胞藻 PCC6803 生物膜的简单方法，用于电子显微镜和功能研究
DOI：
10.1371/journal.pone.0236842
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发表时间：
2020
期刊：
影响因子：
3.7
通讯作者：
I. Vass
中科院分区：
文献类型：
--
作者：
I. Mallick;Prithwiraj Kirtania;M. Szabó;F. Bashir;I. Domonkos;P. Kós;I. Vass
蓝藻在自然界中能形成生物膜，具有重要的生态学意义和应用前景。为了研究它们，我们需要包含健康细胞的生物膜模型，并且可以承受结构研究所需的物理操作。目前，由于缺乏合适的模型系统，对无菌蓝藻生物膜结构和生理特性的综合研究还很有限。在这里，我们提出了一个简单的方法来建立生物膜使用蓝藻集胞藻PCC 6803在标准的实验室条件下直接用于光合活性测量和扫描电子显微镜（SEM）。我们发现，玻璃微纤维过滤器（GMF）与有点粗糙的表面特征提供了一个合适的骨架形成集胞藻PCC6803生物膜。由于非常脆弱，未经处理的GMF无法承受SEM所需的加工步骤。因此，我们使用聚羟基丁酸酯涂层来稳定滤器。我们发现，多达五个涂层导致GMF稳定化，并有可能获得表面附着细胞和广泛的胞外多糖和皮利网络的结构的高分辨率SEM图像，这是生物膜形成的基本特征。通过使用脉冲幅度调制的可变叶绿素荧光成像，也证明了生物膜包含光合活性细胞。因此，集胞藻PCC6803生物膜形成的涂层GMF可用于结构和功能的研究。这里提出的模型很容易复制，并具有高通量研究的潜力。
Cyanobacteria can form biofilms in nature, which have ecological roles and high potential for practical applications. In order to study them we need biofilm models that contain healthy cells and can withstand physical manipulations needed for structural studies. At present, combined studies on the structural and physiological features of axenic cyanobacterial biofilms are limited, mostly due to the shortage of suitable model systems. Here, we present a simple method to establish biofilms using the cyanobacterium Synechocystis PCC6803 under standard laboratory conditions to be directly used for photosynthetic activity measurements and scanning electron microscopy (SEM). We found that glass microfiber filters (GMF) with somewhat coarse surface features provided a suitable skeleton to form Synechocystis PCC6803 biofilms. Being very fragile, untreated GMFs were unable to withstand the processing steps needed for SEM. Therefore, we used polyhydroxybutyrate coating to stabilize the filters. We found that up to five coats resulted in GMF stabilization and made possible to obtain high resolution SEM images of the structure of the surface-attached cells and the extensive exopolysaccharide and pili network, which are essential features of biofilm formation. By using pulse-amplitude modulated variable chlorophyll fluorescence imaging, it was also demonstrated that the biofilms contain photosynthetically active cells. Therefore, the Synechocystis PCC6803 biofilms formed on coated GMFs can be used for both structural and functional investigations. The model presented here is easy to replicate and has a potential for high-throughput studies.
影响因子：
5.4
作者：
Yu Y;You L;Liu D;Hollinshead W;Tang YJ;Zhang F
通讯作者：
Zhang F