title: The Isolation of Plant Organelles and Structures in the Post-genomic Era.
abstract: In this chapter, we provide an overview of the techniques and approaches used in the isolation of plant organelles and structures. This overview shows there is a great diversity of methods currently used for the initial physical disruption of plant tissue before the downstream isolation of a target cellular component. These include hand grinding, high-speed mechanical disruption, and enzymatic digestion of cell walls by a variety of methods. Coupled to these disruption techniques is a wide array of additives included as ingredients in extraction solutions to minimize chemical or physical damage that may occur to target components. These additives are collated into a table outlining their function. We also provide an introduction to some of the history of common approaches used for the isolation plant organelles and structures and a synopsis of the methods used by researchers for assessment of the purity of their isolated structures. This chapter therefore provides an introduction to the following chapters that document the methodology for the isolation of individual plant organelles or structures.
authors.affiliation: The ARC Centre of Excellence in Plant Energy Biology, The University of Western Australia, M316, 35 Stirling Highway, Crawley, Perth, WA, 6009, Australia.
authors.forename: A Harvey
authors.initials: AH
authors.lastname: Millar
authors.affiliation: The ARC Centre of Excellence in Plant Energy Biology and School of Chemistry and Biochemistry, The University of Western Australia, M316, 35 Stirling Highway, Crawley, Perth, WA, 6009, Australia. nicolas.taylor@uwa.edu.au.
authors.forename: Nicolas L
authors.initials: NL
authors.lastname: Taylor
mesh_terms: D002458:Cell Fractionation / Q000295:instrumentation / Q000379:methods*; D002498:Centrifugation / Q000295:instrumentation / Q000379:methods*; D003902:Detergents / Q000737:chemistry; D005638:Fruit / Q000737:chemistry; D023281:Genomics / Q000379:methods*; D015388:Organelles / Q000737:chemistry* / Q000648:ultrastructure; D018517:Plant Roots / Q000737:chemistry; D010944:Plants / Q000737:chemistry*; D011480:Protease Inhibitors / Q000737:chemistry; D036226:Seedlings / Q000737:chemistry; D012639:Seeds / Q000737:chemistry
chemical_list: D003902:Detergents; D011480:Protease Inhibitors
keywords: Cellular disruption; Charge- and affinity-based separation; Density gradient centrifugation; Differential centrifugation; Organelle; Purity assessment; Subcellular compartment; Subcellular structures