Block Maryse A, Albrieux Catherine
Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Institut National Recherche Agronomique, Commissariat à l'Energie Atomique et aux Energies Alternatives, CEA Grenoble, UMR5168, Université Grenoble Alpes, Grenoble, France.
Methods Mol Biol. 2018;1829:123-135. doi: 10.1007/978-1-4939-8654-5_8.
Chloroplasts are specific organelles of plant cells dedicated to photosynthesis and delimited by a two-membrane chloroplast envelope. Their photosynthetic function is based on the development of an operational large internal membrane network, called the thylakoids, and on enzymatic processes present in the chloroplast matrix, called the stroma. Thylakoid membranes are clearly different from the chloroplast envelope and their biogenesis is dependent on biosynthetic and transport activities specific of the chloroplast envelope. Starting with the isolation of intact chloroplasts, the method presents the separation by differential centrifugation of the three main compartments of the chloroplast: the stroma, the thylakoids, and the chloroplast envelope. Three different protocols are provided, adapted for starting leaves of spinach, Arabidopsis, and pea.
叶绿体是植物细胞中专门用于光合作用的特定细胞器，由两层膜组成的叶绿体被膜界定。它们的光合功能基于一个称为类囊体的可运作的大型内膜网络的发育，以及叶绿体基质（称为间质）中存在的酶促过程。类囊体膜与叶绿体被膜明显不同，其生物发生依赖于叶绿体被膜特有的生物合成和转运活动。该方法从完整叶绿体的分离开始，通过差速离心法分离叶绿体的三个主要部分：间质、类囊体和叶绿体被膜。提供了三种不同的方案，适用于菠菜、拟南芥和豌豆的起始叶片。
