Plant Biotechnology, Faculty of Biology, University of Freiburg, Schaenzlestr 1, 79104 Freiburg, Germany.
Plant Cell Rep. 2011 Feb;30(2):205-15. doi: 10.1007/s00299-010-0935-4. Epub 2010 Oct 20.
The moss Physcomitrella patens is increasingly being used as a model for plant systems biology studies. While genomic and transcriptomic resources are in place, tools and experimental conditions for proteomic studies need to be developed. In the present study we describe a rapid and efficient protocol for the simultaneous isolation of chloroplasts and mitochondria from moss protonema. Routinely, 60-100 μg mitochondrial and 3-5 mg chloroplast proteins, respectively, were obtained from 20 g fresh weight of green moss tissue. Using 14 plant compartment marker antibodies derived from seed plant and algal protein sequences, respectively, the evolutionary conservation of the compartment marker proteins in the moss was demonstrated and purity and intactness of the extracted organelles confirmed. This isolation protocol and these validated compartment markers may serve as basis for sub-cellular proteomics in P. patens and other mosses.
藓类植物 Physcomitrella patens 越来越多地被用作植物系统生物学研究的模型。虽然已经有基因组和转录组资源，但仍需要开发蛋白质组学研究的工具和实验条件。在本研究中，我们描述了一种从藓类原丝体中同时分离叶绿体和线粒体的快速、有效的方法。通常，从 20 克新鲜绿色藓类组织中可分别获得 60-100 μg 的线粒体蛋白和 3-5 mg 的叶绿体蛋白。使用分别来自种子植物和藻类蛋白序列的 14 种植物区室标记抗体，证明了区室标记蛋白在藓类植物中的进化保守性，并证实了提取细胞器的纯度和完整性。该分离方案和经过验证的区室标记物可以作为 P. patens 和其他藓类植物亚细胞蛋白质组学的基础。
