An exhaustive qualitative and quantitative profiling of the photosynthetic glycerolipids in three strains of the marine diatom Skeletonema sp. was carried out by ultra performance liquid chromatography-electrospray ionizationquadrupole-time of flight-mass spectrometry. In the diatom thylakoid membrane, monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) account for about 45-70% and 5-15% of the total membrane lipids, respectively. The anionic sulfoquinovosyldiacylglycerol (SQDG) as well as the likewise anionic phosphatidylglycerol (PG) contribute between 10-40% and 4-10% each. The predominant species of MGDG were those with C 16:3 /C 16:3 , C 20:5 / 16:1 , and C 20:5 /C 16:3 . Three main molecular species of DGDG contained C 20:5 /C 16:1 , C 20:5 /C 16:2 , and C 16:1 /C 16:1 . The major molecular species of SQDG were those containing combinations of C 14:0 /C 14:0 , C 14:0 /C 16:0, C 14:0 /C 16:1 , and C 14:0 /C 16:3 . All the PG classes contained the C 18:1 /C 18:1 as the main molecular species. Based on the fatty acid species in sn-2 position, it is indicated that MGDG and DGDG are biosynthesized through prokaryotic pathway exclusively within the chloroplast, whereas PG and SQDG have a typical mixed biosynthetic pathway (both prokaryotic pathway and eukaryotic pathways). The chemical characteristics of photosynthetic glycerolipids related with ecological physiology are discussed.
Phytohormones have attracted wide attention due to their important biological functions. However, their detection is still a challenge because of their complex composition, low abundance and diverse sources. In this study, a novel method of high-performance liquid chromatography with electrospray ionization tandem mass spectrometry was developed and validated for the simultaneous determination of ten phytohormones including indole-3-acetic acid, isopentenyladenine, isopentenyl adenosine, trans-zeatin riboside, zeatin, strigolactones, abscisic acid, salicylic acid, gibberellin A3, and jasmonic acid in Sargassum horneri (S. horneri). The phytohormones were extracted from freeze-dried S. horneri with methanol/water/methanoic acid (15:4:1, v/v/v) analyzed on a Hypersil Gold C18 column and detected by electrospray ionization tandem triple quadrupole mass spectrometry in the multiple reaction monitoring mode. The experimental conditions for the extraction and analysis of phytohormones were optimized and validated in terms of reproducibility, linearity, sensitivity, recovery, accuracy, and stability. Distributions of the phytohormones in the stems, blades, and gas bladder of the S. horneri in drift, fixed, and semi-fixed growing states were investigated for the first time. The observed contents of the phytohormones in S. horneri range from not detected to 5066.67 ng/g (fresh weight). Most phytohormones are distributed mainly in the stems of S. horneri in drift and semi-fixed states.
As an unusual economically important aquaculture species, Sinonovacula constricta possesses high levels of long-chain polyunsaturated fatty acids (LC-PUFA). Previously, our group identified fatty acyl desaturases (Fad) with Δ5 and Δ6 activities in S. constricta, which was the first report of Δ6 Fad in a marine mollusc. Here, we further successfully characterize elongases of very long-chain fatty acids (Elovl) in this important bivalve species, including one Elovl2/5, two Elovl4 isoforms (a and b) and a novel Elovl (c) with Elovl4 activity. In addition, we also determined the desaturation activity of S. constricta Δ6 Fad toward 24:5n-3 to give 24:6n-3, a key intermediate in docosahexaenoic acid (DHA) biosynthesis. Therefore, S. constricta is the first marine mollusc reported to possess all Fad and Elovl activities required for LC-PUFA biosynthesis via the 'Sprecher pathway'. This finding greatly increases our understanding of LC-PUFA biosynthesis in marine molluscs. Phylogenetic analysis by interrogating six marine molluscan genomes, and previously functionally characterized Elovl and Fad from marine molluscs, suggested that DHA biosynthetic ability was limited to a few species, due to the general lack of Δ4 or Δ6 Fad in most molluscs.
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