Plant Methods volume 20 , Article number: 38 Cite this article. Metrics details. The extraction of thylakoids is an essential step in studying the structure of photosynthetic complexes and several other aspects of the photosynthetic process in plants. Conventional protocols have been developed for selected land plants grown in controlled conditions. Plants accumulate defensive chemical compounds such as polyphenols to cope with environmental stresses.
When the polyphenol levels are high, their oxidation and cross-linking properties prevent thylakoid extraction. In this study, we developed a method to counteract the hindering effects of polyphenols by modifying the grinding buffer with the addition of both vitamin C VitC and polyethylene glycol PEG This protocol was first applied to the marine plant Posidonia oceanica and then extended to other plants synthesizing substantial amounts of polyphenols, such as Quercus pubescens oak and Vitis vinifera grapevine.
Moreover, total protein extraction from frozen P. Our work shows that the use of PEG and VitC significantly improves the isolation of native thylakoids, native photosynthetic complexes, and total proteins from plants containing high amounts of polyphenols and thus enables studies on photosynthesis in various plant species grown in natural conditions.
The ecological crisis is accelerating the need to study photosynthesis in free-growing wild plants to complement data obtained on plants grown under controlled laboratory conditions. Indeed, the latter conditions do not consider the ecological and evolutionary relevance of the underlying photosynthetic mechanisms.
For example, studies of marine plants rarely address the response mechanisms to a light environment that strongly differ depending on the depth, at which the plants live 1โ40 m. Extraction of thylakoid membranes and photosynthetic protein complexes is crucial to performing such studies. However, thylakoid extraction protocols have been optimized for model plants grown in laboratory conditions, such as Arabidopsis thaliana or Pisum sativum [ 1 , 2 , 3 ], and proved very inefficient in extracting membranes from wild plants and crops like oak and grapevine.
