Bag Pushan, Schröder Wolfgang P, Jansson Stefan, Farci Domenica
Department of Plant Physiology, Umeå Plant Science Centre, Umeå University, Sweden.
Department of Chemistry, Umeå University, Sweden.
Bio Protoc. 2021 Sep 5;11(17):e4144. doi: 10.21769/BioProtoc.4144.
Photosynthesis is the main process by which sunlight is harvested and converted into chemical energy and has been a focal point of fundamental research in plant biology for decades. In higher plants, the process takes place in the thylakoid membranes where the two photosystems (PSI and PSII) are located. In the past few decades, the evolution of biophysical and biochemical techniques allowed detailed studies of the thylakoid organization and the interaction between protein complexes and cofactors. These studies have mainly focused on model plants, such as , pea, spinach, and tobacco, which are grown in climate chambers even though significant differences between indoor and outdoor growth conditions are present. In this manuscript, we present a new mild-solubilization procedure for use with "fragile" samples such as thylakoids from conifers growing outdoors. Here, the solubilization protocol is optimized with two detergents in two species, namely Norway spruce () and Scots pine (). We have optimized the isolation and characterization of PSI and PSII multimeric mega- and super-complexes in a close-to-native condition by Blue-Native gel electrophoresis. Eventually, our protocol will not only help in the characterization of photosynthetic complexes from conifers but also in understanding winter adaptation.
光合作用是捕获阳光并将其转化为化学能的主要过程，几十年来一直是植物生物学基础研究的重点。在高等植物中，该过程发生在类囊体膜上，两个光系统（PSI和PSII）位于其中。在过去几十年中，生物物理和生化技术的发展使得对类囊体组织以及蛋白质复合物与辅因子之间的相互作用能够进行详细研究。这些研究主要集中在模式植物上，如豌豆、菠菜和烟草，它们是在气候室中种植的，尽管室内和室外生长条件存在显著差异。在本论文中，我们提出了一种新的温和增溶方法，用于处理“易碎”样品，如来自户外生长的针叶树的类囊体。在这里，增溶方案在两种植物中用两种去污剂进行了优化，即挪威云杉（Picea abies）和欧洲赤松（Pinus sylvestris）。我们通过蓝色非变性凝胶电泳在接近天然的条件下优化了PSI和PSII多聚体巨型和超级复合物的分离与表征。最终，我们的方案不仅将有助于表征针叶树的光合复合物，还将有助于理解冬季适应性。
