CI-976

Phosphatidic acid (PA) and lysophosphatidic acid acyltransferases (LPAATs) might be critical for the secretory pathway. Four extra-plastidial LPAATs (LPAAT2, 3, 4, and 5) were identified in Arabidopsis thaliana. These AtLPAATs display a specific enzymatic activity converting lysophosphatidic acid to PA and are located in the endomembrane system. We investigate a putative role for AtLPAATs 3, 4, and 5 in the secretory pathway of root cells through genetical (knockout mutants), biochemical (activity inhibitor, lipid analyses), and imaging (live and immuno-confocal microscopy) approaches. Treating a lpaat4;lpaat5 double mutant with the LPAAT inhibitor CI976 produced a significant decrease in primary root growth. The trafficking of the auxin transporter PIN2 was disturbed in this lpaat4;lpaat5 double mutant treated with CI976, whereas trafficking of H+-ATPases was unaffected. The lpaat4;lpaat5 double mutant is sensitive to salt stress, and the trafficking of the aquaporin PIP2;7 to the plasma membrane in the lpaat4;lpaat5 double mutant treated with CI976 was reduced. We measured the amounts of neo-synthesized PA in roots, and found a decrease in PA only in the lpaat4;lpaat5 double mutant treated with CI976, suggesting that the protein trafficking impairment was due to a critical PA concentration threshold.

Environmental monitoring studies based on target analysis capture only a small fraction of contaminants of emerging concern (CECs) and miss pollutants potentially harmful to wildlife. Environmental specimen banks, with their archived samples, provide opportunities to identify new CECs by temporal trend analysis and non-target screening. In this study, archived white-tailed sea eagle (Haliaeetus albicilla) muscle tissue was analysed by non-targeted high-resolution mass spectrometry. Univariate statistical tests (Mann-Kendall and Spearman rank) for temporal trend analysis were applied as prioritisation methods. A workflow for non-target data was developed and validated using an artificial time series spiked at five levels with gradient concentrations of selected CECs (n = 243). Pooled eagle muscle tissues collected 1965-2017 were then investigated with an eight-point time series using the validated screening workflow. Following peak detection, peak alignment, and blank subtraction, 14 409 features were considered for statistical analysis. Prioritisation by time-trend analysis detected 207 features with increasing trends. Following unequivocal molecular formula assignment to prioritised features and further elucidation with MetFrag and EU Massbank, 13 compounds were tentatively identified, of which four were of anthropogenic origin. These results show that it is possible to prioritise new CECs in archived biological samples using univariate statistical approaches.