Sweetlove Lee J, Taylor Nicolas L, Leaver Christopher J
Department of Plant Sciences, University of Oxford, United Kingdom.
Methods Mol Biol. 2007;372:125-36. doi: 10.1007/978-1-59745-365-3_9.
The ability to isolate intact, functional mitochondria from plant tissues is a key technique in the study of the genome, proteome, and metabolic function of the plant mitochondrion. Traditionally, mitochondrial plant researchers have turned to specific plant systems and organs (such as potato tubers and pea shoots) from which mitochondria are readily isolated in large quantities. However, increasingly, research is focused on a small number of model species, and there is a need to adapt existing protocols to allow the isolation of mitochondria from these model species. Arguably, the most important of these is Arabidopsis thaliana, for which a formidable array of genetic resources is available. However, because of its relatively small size and the absence of large heterotrophic organs, Arabidopsis is a challenging plant from which to isolate mitochondria. Here, we present two methods for isolating mitochondria from Arabidopsis, either from heterotrophic cell suspension cultures or from hydroponic seedling cultures. We also present details of commonly used assays to assess the physical and functional integrity of the isolated organelles.
从植物组织中分离完整、有功能的线粒体的能力是研究植物线粒体基因组、蛋白质组和代谢功能的一项关键技术。传统上，植物线粒体研究人员会选用特定的植物系统和器官（如马铃薯块茎和豌豆苗），从中很容易大量分离出线粒体。然而，越来越多的研究集中在少数模式物种上，因此需要调整现有方案，以便从这些模式物种中分离线粒体。可以说，其中最重要的是拟南芥，它拥有大量丰富的遗传资源。然而，由于其相对较小的尺寸以及缺乏大型异养器官，从拟南芥中分离线粒体具有挑战性。在这里，我们介绍两种从拟南芥中分离线粒体的方法，一种是从异养细胞悬浮培养物中分离，另一种是从水培幼苗培养物中分离。我们还详细介绍了常用的检测方法，以评估分离出的细胞器的物理完整性和功能完整性。
