Comparison of methods for extracting thylakoid membranes of Arabidopsis plants.

Physiologia Plantarum
Yang-Er ChenW P Schroder

Abstract

Robust and reproducible methods for extracting thylakoid membranes are required for the analysis of photosynthetic processes in higher plants such as Arabidopsis. Here, we compare three methods for thylakoid extraction using two different buffers. Method I involves homogenizing the plant material with a metal/glass blender; method II involves manually grinding the plant material in ice-cold grinding buffer with a mortar and method III entails snap-freezing followed by manual grinding with a mortar, after which the frozen powder is thawed in isolation buffer. Thylakoid membrane samples extracted using each method were analyzed with respect to protein and chlorophyll content, yields relative to starting material, oxygen-evolving activity, protein complex content and phosphorylation. We also examined how the use of fresh and frozen thylakoid material affected the extracts' contents of protein complexes. The use of different extraction buffers did not significantly alter the protein content of the extracts in any case. Method I yielded thylakoid membranes with the highest purity and oxygen-evolving activity. Method III used low amounts of starting material and was capable of capturing rapid phosphorylation changes in the sample at ...Continue Reading

References

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Mar 10, 2015·Annual Review of Biochemistry·Nathan Nelson, Wolfgang Junge

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Citations

Sep 8, 2018·Plant & Cell Physiology·Kristen Van GelderTariq A Akhtar
Feb 16, 2020·International Journal of Molecular Sciences·Yang-Er ChenShu Yuan
Jan 8, 2020·International Journal of Molecular Sciences·Yanger ChenHuaiyu Zhang
Aug 28, 2020·Journal of Experimental Botany·Tim JiangRongmin Zhao
Sep 17, 2021·Physiologia Plantarum·Yang-Er ChenWolfgang P Schröder

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