Abstract
Plant phospholipids can be produced in the endoplasmic reticulum or plastids. Lipids from different sources can be distinguished by the fatty acid profile, in terms of the preferred fatty acid species esterified to the sn-1 or sn-2 position of the glycerol backbone (Ohlrogge and Browse, 1995). This protocol is used to determine the fatty acid profile in total plant phospholipids by the treatment of sn-2 specific phospholipase A2 (PLA2).
Keywords: Lipidomics, Phospholipase A2, Prokaryotic pathway, Eukaryotic pathway
Materials and Reagents
Equipment
Procedure
Representative data
Representative data from Liu et al., 2015 (Figure 1). Total lipids were digested by PLA2, followed by analysis of PC and LysoPC by mass spectrometry. The increase of LPC16 in digested sample and increase of PC34 in undigested samples in RNAi lines compared with WT are similar, suggesting the increased PC34 in RNAi lines have C16 fatty acid esterified to the sn-1 position, which originates from the eukaryotic pathway. Figure 1. Analysis of fatty acyl chain position/composition in phosphatidylcholine (PC). A. Content of LysoPC with C16 and C18 fatty acids after PLA2 treatments of lipids from two AAPT RNAi lines and WT plants. B. Content of PC with C34 and C36 fatty acids without PLA2 treatments of lipids from two AAPT RNAi lines and WT plants. * and ** mark differences between WT and mutant at P < 0.05 and at P < 0.01, respectively based on Student’s t-test. Values are means ± SD (n = 3).
Recipes
Acknowledgments
This protocol is adapted from the work of Williams et al., 1995. Work was supported by the National Institute of Food and Agriculture Grant 2016-67013-24429 and the U.S. Department of Energy (DOE) Grant DE-AR0000202.
References
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.