Schober Alexander F, Flori Serena, Finazzi Giovanni, Kroth Peter G, Bártulos Carolina Río
Plant Ecophysiology, Department of Biology, University of Konstanz, Konstanz, Germany.
Department of Cell and Molecular Processes, Marine Biological Association of the United Kingdom, Plymouth, UK.
Methods Mol Biol. 2018;1829:189-203. doi: 10.1007/978-1-4939-8654-5_13.
The so-called "complex" plastids from diatoms possessing four envelope membranes are a typical feature of algae that arose from secondary endosymbiosis. Studying isolated plastids from these algae may allow answering a number of fundamental questions regarding diatom photosynthesis and plastid functionality. Due to their complex architecture and their integration into the cellular endoplasmic reticulum (ER) system, their isolation though is still challenging. In this work, we report a reliable isolation technique that is applicable for the two model diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum. The resulting plastid-enriched fractions are of homogenous quality, almost free from cellular contaminants, and feature structurally intact thylakoids that are capable to perform oxygenic photosynthesis, though in most cases they seem to lack most of the stromal components as well as plastid envelopes.
具有四层包膜的硅藻所谓的“复杂”质体是藻类次生内共生起源的典型特征。研究从这些藻类中分离出的质体可能有助于回答一些关于硅藻光合作用和质体功能的基本问题。然而，由于它们复杂的结构以及与细胞内质网（ER）系统的整合，其分离仍然具有挑战性。在这项工作中，我们报告了一种可靠的分离技术，该技术适用于两种模式硅藻——三角褐指藻和假微型海链藻。得到的富含质体的组分质量均匀，几乎没有细胞污染物，并且具有结构完整的类囊体，能够进行放氧光合作用，不过在大多数情况下，它们似乎缺少大部分的基质成分以及质体包膜。
