Domozych David, Lietz Anna, Patten Molly, Singer Emily, Tinaz Berke, Raimundo Sandra C
Department of Biology, Skidmore College, Saratoga Springs, NY, 12866, USA.
Methods Mol Biol. 2017;1563:91-105. doi: 10.1007/978-1-4939-6810-7_7.
The unicellular green alga, Penium margaritaceum, represents a novel and valuable model organism for elucidating cell wall dynamics in plants. This organism's cell wall contains several polymers that are highly similar to those found in the primary cell walls of land plants. Penium is easily grown in laboratory culture and is effectively manipulated in various experimental protocols including microplate assays and correlative microscopy. Most importantly, Penium can be live labeled with cell wall-specific antibodies or other probes and returned to culture where specific cell wall developmental events can be monitored. Additionally, live cells can be rapidly cryo-fixed and cell wall surface microarchitecture can be observed with variable pressure scanning electron microscopy. Here, we describe the methodology for maintaining Penium for experimental cell wall enzyme studies.
单细胞绿藻Margaritaceum念珠藻是阐明植物细胞壁动态变化的一种新型且有价值的模式生物。该生物的细胞壁包含几种聚合物，这些聚合物与陆地植物初生细胞壁中的聚合物高度相似。念珠藻易于在实验室培养中生长，并且在包括微孔板分析和相关显微镜检查在内的各种实验方案中都能有效操作。最重要的是，念珠藻可以用细胞壁特异性抗体或其他探针进行活细胞标记，然后放回培养物中，在那里可以监测特定的细胞壁发育事件。此外，活细胞可以快速冷冻固定，并且可以用可变压力扫描电子显微镜观察细胞壁表面微观结构。在这里，我们描述了用于维持念珠藻以进行实验性细胞壁酶研究的方法。
