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Summary: Fucoxanthin, a natural carotenoid that has substantial pharmaceutical value due to its anticancer, antioxidant, antiobesity, and antidiabetic properties, is biosynthesized from glyceraldehyde‐3‐phosphate (G3P) via a series of enzymatic reactions. However, our understanding of the transcriptional mechanisms involved in fucoxanthin biosynthesis remains limited.Using reverse genetics, the med8 mutant was identified based on its phenotype of reduced fucoxanthin content, and the biological functions of MED8 in fucoxanthin synthesis were characterized using approaches such as gene expression, protein subcellular localization, protein–protein interaction and chromatin immunoprecipitation assay.Gene‐editing mutants of MED8 exhibited decreased fucoxanthin content as well as reduced expression levels of six key genes involved in fucoxanthin synthesis, namely DXS, PSY1, ZDS‐like, CRTISO5, ZEP1, and ZEP3, when compared to the wild‐type (WT) strain. Furthermore, we showed that MED8 interacts with HSF3, and genetic analysis revealed their shared involvement in the genetic pathway governing fucoxanthin synthesis. Additionally, HSF3 was required for MED8 association with the promoters of the six fucoxanthin synthesis genes.In conclusion, MED8 and HSF3 are involved in fucoxanthin synthesis by modulating the expression of the fucoxanthin synthesis genes. Our results increase the understanding of the molecular regulation mechanisms underlying fucoxanthin synthesis in the diatom P. tricornutum. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) is one of the major contaminants in aquatic ecosystem. Stearoyl‐coenzyme A desaturase 1 (Scd1) has been implicated in adaptive responses to environmental stressors. The objectives of this study are (a) to characterize scd1 mRNA from silver pomfret (Pampus argenteus); (b) to investigate the expression and activity of Scd1 in silver pomfret exposed to Cd; and (c) to investigate how Cd modifies scd1 gene transcription in silver pomfret. Results indicated that Scd1 was generally conserved across fish species and scd1 mRNA level was higher by far in the brain and liver, followed by the kidney and intestine. Exposure to Cd led to significant changes of the expression and activity of Scd1 in in the liver and intestine. The liver mRNA abundance of scd1 was significantly lower in the Cd‐treated groups than in the control group. The 10 days treatment with 1 mg/L Cd significantly upregulated the intestinal scd1 mRNA level, an approximately 9‐fold higher in the 1 mg/L Cd‐treated group as compared with the control group. Accordingly, Scd1 activity indices (18:1n‐9/18:0) in the liver were significantly decreased in the 0.5 mg/L group compared with the control group, while Scd1 activity indices in the intestine were significantly increased in the 1 mg/L group compared with the control group. Moreover, overexpression of sterol regulatory element binding transcription factor 1 (Srebp1) and peroxisome proliferator–activated receptor γ (Pparγ)in HEK 293T cells produced a 2‐fold increment in the activity of the scd1 promoter. Furthermore, srebp1 had a similar expression pattern to scd1 in the liver and intestine of silver pomfret exposed to Cd. These results indicated that Cd could regulate scd1 expression, possibly through the transcriptional factor Srebp1. [ABSTRACT FROM AUTHOR]

SUMMARY: The small athecate dinoflagellate Karlodinium zhouanum is a species recently described in the coastal waters of China. K. zhouanum is morphologically similar to Karlodinium veneficum, a typical ichthyotoxic blooming karlotoxin‐producing species, and it is impossible to distinguish between these two species based on light microscopy. In this study, strains of K. zhouanum isolated from the East China Sea were studied. By analyzing toxins, toxicity, lipid characteristics and typical molecular and physiological traits of this species, K. zhouanum was shown to be nontoxic to brine shrimp and widely spread over the coastal waters of China. No karlotoxin‐like toxin was detected by liquid chromatography‐mass spectrometry (LC–MS). Instead of gymnodinosterol, the critical sterol in toxic K. veneficum, 27(nor)‐24S‐4α‐Methyl‐5α‐ergosta‐8(14)‐en‐3β‐ol (NEE) was dominant in K. zhouanum, while gymnodinosterol was absent. These sterol characteristics may provide not only support for the species separation between toxic and nontoxic species of Karlodinium but also environmental survey tools to differentiate the contribution of nontoxic Karlodinium strains, which has been unclear until now. [ABSTRACT FROM AUTHOR]

Bivalves, a highly diverse and the most evolutionarily successful class of invertebrates native to aquatic habitats, provide valuable molecular resources for understanding the evolutionary adaptation and aquatic ecology. Here, we reported a high‐quality chromosome‐level genome assembly of the razor clam Sinonovacula constricta using Pacific Bioscience single‐molecule real‐time sequencing, Illumina paired‐end sequencing, 10X Genomics linked‐reads and Hi‐C reads. The genome size was 1,220.85 Mb, containing scaffold N50 of 65.93 Mb and contig N50 of 976.94 Kb. A total of 899 complete (91.92%) and seven partial (0.72%) matches of the 978 metazoa Benchmarking Universal Single‐Copy Orthologs were determined in this genome assembly. And Hi‐C scaffolding of the genome resulted in 19 pseudochromosomes. A total of 28,594 protein‐coding genes were predicted in the S. constricta genome, of which 25,413 genes (88.88%) were functionally annotated. In addition, 39.79% of the assembled genome was composed of repetitive sequences, and 4,372 noncoding RNAs were identified. The enrichment analyses of the significantly expanded and contracted genes suggested an evolutionary adaptation of S. constricta to highly stressful living environments. In summary, the genomic resources generated in this work not only provide a valuable reference genome for investigating the molecular mechanisms of S. constricta biological functions and evolutionary adaptation, but also facilitate its genetic improvement and disease treatment. Meanwhile, the obtained genome greatly improves our understanding of the genetics of molluscs and their comparative evolution. [ABSTRACT FROM AUTHOR]

SUMMARY: Lipid metabolites play an important role in understanding the stress physiology of Pyropia haitanensis, and can be used to facilitate development of stress‐resistant Pyropia cultivars. Therefore, in this study ultra performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (UPLC‐Q‐TOF‐MS) and gas chromatography–mass spectrometry (GC–MS) based metabolomics approaches were developed to screen the responses of lipid metabolites such as phospholipids, glycolipids, fatty acids and volatile organic compounds (VOCs) to different heat shock times. A total of 26 potential lipid biomarkers including Lyso‐monogalactosyldiacylglycerol (Lyso‐MGDG), Lyso‐digalactosyldiacylglycerol (Lyso‐DGDG), sulfoquinovosylmonoacylglycerols (SQMG), sulfoquinovosyldiacylglycerol (SQDG), diacylglyceryltrimethylhomoserine (DGTS), triacylglycerol (TAG), Lyso‐phosphatidicacid (Lyso‐PA), Lyso‐phosphatidylcholine (Lyso‐PC), Lyso‐phosphatidylethanolamine (Lyso‐PE), Lyso‐phosphatidylglycerol (Lyso‐PG), phosphatidylglycerol (PG), phosphatidylinositol (PI), and phosphatidylinositol phosphate (PIP) were identified, most of which responded to high temperature by reducing or increasing levels after stimulation for 1 h or 6 h. After times longer than 6 h, the levels of most lipids gradually recovered to the control group levels. Moreover, the balance of lipids and fatty acids transformation was disrupted. Overall, 11 total fatty acids (TFAs), 13 free fatty acids (FFAs) and 29 VOCs were identified during 0–72 h of high temperature stress. The FFAs, especially polyunsaturated C 20 fatty acids and VOCs, showed opposing change trends, indicating the transformation between C 20 fatty acids and VOCs. Overall, this study provides important insights into the metabolic variations of P. haitanensis under different heat shock time and the relationship between the conversion of lipids, fatty acids, and VOCs. The information provided herein will facilitate efficient development and improvement of Pyropia quality by producing cultivars resistant to high temperature. [ABSTRACT FROM AUTHOR]

Abstract: BACKGROUND: Steryl glycosides (SGs) are sterol conjugates found in various plants, especially in those making up human diets. It has been demonstrated that SGs have potential health benefits, and they could be used as food supplements in a variety of food matrixes. Marine microalgae are a potential resource for human food and ingredients. In this study, gas chromatography–triple quadrupole mass spectrometry (GC–QQQ‐MS) was used to characterize unknown SGs in eight microalgae belonging to different classes (Isochrysis galbana 3011, Pavlova viridis, Platymonas helgolandica, Conticribra weissflogii, Thalassiosira pseudonana, Nitzschia closterium, Gymnodinium sp., and Karlodinum veneficum). RESULTS: The SGs were first extracted from lyophilized algae with chloroform–methanol, purified by solid‐phase extraction and analyzed as trimethylsilyl derivatives. Nine SGs have been identified. In particular, new SGs like occelasteryl glycoside and stellasteryl glycoside were found in Gymnodinium sp., 24‐methylene cholesteryl glycoside was detected in P. helgolandica, and 4,24‐dimethylcholestan‐3‐yl glycoside was identified as the main constituent of microalga K. veneficum. The results also showed that the compositions of SGs in different microalgae varied, with a range of 5.234 to 0.036 g kg−1, and microalga P. viridis contained the most abundant SGs. CONCLUSION: GC–QQQ‐MS is a powerful tool to detect SGs with different structures from a variety of microalgae. The compositions of SGs in different microalgae varied greatly. Microalgae are a good source of highly valued SGs. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

BACKGROUND Sinonovacula constricta is an economically and nutritionally important bivalve native to the estuaries and mudflats of China, Japan and Korea. In the present study, S. constricta, cultured either under experimental conditions or collected directly from natural coastal areas with different seawater salinities, was investigated for changes in proximates, amino acids and lipids. RESULTS When culture salinity was increased, levels of moisture, carbohydrate, crude protein and crude lipid were significantly decreased, whereas the level of ash was significantly increased. The level of Ala was increased by 1.5- to 2-fold, whereas the contents of most lipids were significantly decreased, and the proportion of phosphatidylethanolamine was significantly increased. Notably, a high proportion of ceramide aminoethylphosphonates was detected in S. constricta reared at all salinities. The energy content appears to be higher in S. constricta reared at higher salinity. In experimental S. constricta, when the salinity was enhanced, the changes of compositions were very close to those reared at constant high salinity. CONCLUSION Sinonovacula constricta reared at higher salinities possesses a superior quality. A short period of exposure to a higher salinity for farmed S. constricta reared at a lower salinity before harvest would be useful with respect to improving its nutritive value. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Rationale Diacylgycerol- N-trimethylhomoserine (DGTS) and diacylglycerylhydroxymethyl- N,N,N-trimethyl-β-alanine (DGTA) are structural isomers that are the most commonly described betaine lipids in microalgae. The structural differentiation and precise identification of DGTS and DGTA in microalgae need to be established during mass spectrometry analysis. Methods Total lipid was extracted from Amphora spp. with CHCl3/CH3OH (1:1, v/v). The qualitative analysis of DGTS and DGTA in Amphora spp. was carried out using Li+/H+ dual mode by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry operating in MSE mode (UPLC/QTOF MSE). Results Characteristic fragment ions [C10H22O5N]+ at m/z 236.15 and [C7H14O2N]+ at m/z 144.10 from the [M + H]+ precursor ion can be used for the qualitative analysis of both DGTA and DGTS, whereas the loss of m/z 87 and 74 from the [M + Li]+ precursor ion are specific for DGTS, and the loss of m/z 103 from the [M + Li]+ precursor ion is only for DGTA. As a result, 9 DGTSs and 16 DGTAs with different fatty acids were identified simultaneously in Amphora spp. Semi-quantitative analysis of DGTS and DGTA in Amphora spp. showed that the contents of DGTS ranged from 0.003 to 0.438 nmol mg-1 dw, and that of DGTA from 0.004 to 0.414 nmol mg-1 dw. Conclusions This is the first report to achieve the ambiguous structural identification of DGTS and DGTA by UPLC/QTOF MSE using dual Li+/H+ adduct ion mode, which has remained a challenge in the past. It could provide new insights into their phylogeny and be helpful to characterize the natural phytoplankton communities as intact polar lipid biomarkers. Copyright © 2017 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

BACKGROUND Head space solid-phase microextraction-gas chromatography-mass spectrometry has been applied to analyze the volatile components of six marine microalgae ( Thalassiosira weissflogii, Nitzschia closterium, Chaetoceros calcitrans, Platymonas helgolandica, Nannochloropsis spp . and Dicrateria inornata) from Bacillariophyta, Chlorophyta and Chrysophyta, respectively, in different growth phases. RESULTS All volatile compounds were identified by database searching in the NIST08 Mass Spectral Library and analyzed by principal component analysis with SIMCA-P software (Umetrics, Umea, Sweden). The results clearly revealed that the volatile components of the six microalgae were significantly different in the exponential, stationary and declining phases. Aldehydes, alkanes, some esters and dimethyl sulfide significantly changed in different growth phases. CONCLUSION This is the first report on the comprehensive characteristics of volatile components in different microalgae and in different growth phases. The results may provide reference data for studies on the flavor of cultivated aquatic organism, odor formation in nature water, choice of feeding period and microalgae species selection for the artificial rearing of marine organisms. © 2016 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

SUMMARY The diatom Conticribra weissflogii is a microalga with high nutrition value, rich in docosahexaenoic acids ( DHA) and eicosapentaenoic acids ( EPA). In order to study the effect of culture conditions on the changes of lipid compositions, the intact lipid structural profiles and fatty acids in C. weissflogii were monitored under static and aerated culture conditions. The results showed that, lipids identified in C. weissflogii were neutral lipid triacylglycerols ( TAG), betaine lipid diacylglycerylcarboxy-N-hydroxymethyl-choline ( DGCC), phosphatidylcholine ( PC) and four classes of photosynthetic glycerolipids. The profiles of lipid metabolites of C. weissflogii were different between two culture modes, with the following characteristics under aerated conditions: TAGs increased significantly, whereas the levels of sulfoquinovosyl diacylglycerol ( SQDG), monogalactosyldiacylglycerol ( MGDG), and DGCC decreased. Furthermore, higher contents of EPA-rich TAG and EPA/DHA-rich DGCC were detected at the end of stationary phase, while EPA/DHA-rich PC, EPA-rich MGDG and EPA-rich digalactosyldiacylglycerol ( DGDG) were obtained in the exponential phase under static conditions. Meanwhile, the contents of almost all classes of the essential fatty acids ( EFAs)-enriched lipids increased at onset of stationary phase under aerated conditions. Taken together, given that the high levels of EFAs are required for artificial rearing of marine organisms, aeration is critically important for increasing the production rate and the contents of EFA molecules and therefore increasing the nutritional value of the microalgae. [ABSTRACT FROM AUTHOR]

The shrimp Rimicaris exoculata is the most abundant species in hydrothermal vents. Lipids, the component of membranes, play an important role in maintaining their function normally in such extreme environments. In order to understand the lipid adaptation of R. exoculata (HV shrimp) to hydrothermal vents, we compared its lipid profile with the coastal shrimp Litopenaeus vannamei (EZ shrimp) which lives in the euphotic zone, using ultra performance liquid chromatography electrospray ionization-quadrupole time-of-flight mass spectrometry. As a result, the following lipid adaptation can be observed. (1) The proportion of 16:1 and 18:1, and non-methylene interrupted fatty acid (48.9 and 6.2 %) in HV shrimp was higher than that in EZ shrimp (12.7 and 0 %). While highly-unsaturated fatty acids were only present in the EZ shrimp. (2) Ceramide and sphingomyelin in the HV shrimp were enriched in d14:1 long chain base (96.5 and 100 %) and unsaturated fatty acids (67.1 and 57.7 %). While in the EZ shrimp, ceramide and sphingomyelin had the tendency to contain d16:1 long chain base (68.7 and 75 %) and saturated fatty acids (100 and 100 %). (3) Triacylglycerol content (1.998 ± 0.005 nmol/mg) in the HV shrimp was higher than that in the EZ shrimp (0.092 ± 0.005 nmol/mg). (4) Phosphatidylinositol and diacylglycerol containing highly-unsaturated fatty acids were absent from the HV shrimp. (5) Lysophosphatidylcholine and lysophosphatidylethanolamine were rarely detected in the HV shrimp. A possible reason for such differences was the result of food resources and inhabiting environments. Therefore, these lipid classes mentioned above may be the biomarkers to compare the organisms from different environments, which will be benefit for the further exploitation of the hydrothermal environment. [ABSTRACT FROM AUTHOR]

The two morphologically similar microalgae NMBluh014 and NMBluh-X belong to two different strains of Nannochloropsis oceanica. They possess obviously different feeding effects on bivalves, but are indistinguishable by 18S rRNA and morphological features. In this work, lipidomic analysis followed by principal component analysis and orthogonal projections to latent structures discriminant analysis provided a clear distinction between these strains. Metabolites that definitively contribute to the classification were selected as potential biomarkers. The most important difference in polar lipids were sulfoquinovosyldiacylglycerol (containing 18:1/16:0 and 18:3/16:0) and monogalactosyldiacylglycerol (containing 18:3/16:3 and 20:5/14:0), which were detected only in NMBluh-X. Additionally, an exhaustive qualitative and quantitative profiling of the neutral lipid triacylglycerol ( TAG) in the two strains was carried out. The predominant species of TAG containing 16:1/16:1/16:1 acyl groups was detected only in NMBluh-X with a content of ~93.67 ± 11.85 nmol · mg−1 dry algae at the onset of stationary phase. Meanwhile, TAG containing 16:0/16:0/16:0 was the main TAG in NMBluh014 with a content of 40.25 ± 3.92 nmol · mg−1. These results provided the most straightforward evidence for differentiating the two species. The metabolomic profiling indicated that NMBluh-X underwent significant chemical and physiological changes during the growth process, whereas NMBluh014 did not show such noticeable time-dependent metabolite change. This study is the first using Ultra Performance Liquid Chromatography coupled with Electrospray ionization-Quadrupole-Time of Flight Mass Spectrometry (UPLC-Q-TOF-MS) for lipidomic profiling with multivariate statistical analysis to explore lipidomic differences of plesiomorphous microalgae. Our results demonstrate that lipidomic profiling is a valid chemotaxonomic tool in the study of microalgal systematics. [ABSTRACT FROM AUTHOR]

RATIONALE The precise identification of fatty acids at the sn-2 position of triacylglycerols (TAGs), especially for positional regioisomers (AAB/ABA), needs to be established during mass spectrometry analysis. The detailed structural information about TAGs is significant not only for the assessment of biofuel quality, but also for the tracing of biosynthetic precursors. METHODS Total lipid was extracted from T. pseudonana by a modified Bligh and Dyer method. The qualitative analysis of TAGs in T. pseudonana was carried out using ultra-performance liquid chromatography/electrospray ionization-quadrupole time-of-flight mass spectrometry (UPLC/ESI-Q-TOF-MS). The raw LC/MS data were analyzed using MassLynx software (version 4.1, Waters). RESULTS The acyl group at the sn-2 position of the TAGs has been identified unequivocally by [M + Li-R1/3COOH-R2CH = CHCOOH]+ and the abundance of [M + Li-R1/3COOH-R2CH = CHCOOH]+ can be used to confirm whether the TAG isomers are co-eluted. In total, twelve TAGs were identified in T. pseudonana based on the fragmentation patterns discussed above. The data indicated that only C16 fatty acids were located at the sn-2 position, which was important to trace the biosynthetic precursors of TAGs. CONCLUSIONS We put forward a hypothesis that TAGs in T. pseudonana are only derived from lipids in chloroplasts through prokaryotic biosynthesis pathway based on the precise information of sn-2 fatty acids, which is significant not only for the assessment of biofuel quality, but also for the tracing of biosynthetic precursors. Copyright © 2013 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

ABSTRACT A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the quantitative determination of phytohormones and betaines in algae. The results showed that phytohormones and betaines were separated with high efficiency on Hypersil Gold C18 and Cnwsil SCX columns. Mass spectrometric detection was performed using positive or negative electrospray ionization in selective reaction monitoring mode (SRM). Linearity of the method was good with correlation coefficients (r2) > 0.9951 in the range of 0.005-5 mg/L. The limits of detection were from 0.004 to 0.86 µg/L and the limits of quantification were in the range from 0.01 to 2.8 µg/L for the investigated phytohormones and betaines. The obtained recoveries varied between 61.33 and 90.39%, and the relative standard deviations were <15%. Using the developed methods, seven types of phytohormones and two types of betaines in Laminaria japonica, and seven types of phytohormones and one type of betaine in Pyropia haitanensis, which were collected in Xiangshan, Zhejiang Province, China, were determined. Thus, LC-MS/MS was demonstrated to be a powerful tool for the comprehensive analysis of phytohormones and betaines in algae, owing to its large dynamic range and excellent sensitivity. Copyright © 2013 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

RATIONALE Glycosphingolipids (GSLs) are important components of the cell membrane; however, little is known about GSLs in microalgae. We analyzed GSLs in three strains of Skeletonema microalgae by liquid chromatography coupled with mass spectrometry. METHODS Total lipid was extracted from Skeletonema microalgae. Separation of lipids was achieved via reversed-phase liquid chromatography, and mass spectrometric analysis of the lithium adduct of GSLs was conducted to determine their structures. RESULTS Two types of novel glycosphingolipids were identified from three strains of Skeletonema microalgae. The N-acyl groups were primarily long-chain saturated or monoenoic fatty acids (monounsaturated fatty acids) with 16C and 20 to 24C. The sphingoid long-chain bases (LCB) were sphingosine, sphingadienine and sphingatrienine. The saccharide polar head groups of glycosphingolipids were disaccharide (heptose-hexose) or trisaccharide (heptose-hexose-hexose). Semi-quantitative analysis of GLSs in the three strains of microalgae showed that GSL contents ranged from 0.09 to 8.79 nmol/mg dry microalgae powder. CONCLUSIONS A qualitative method was developed for the identification of GSLs in microalgae. Two types of novel GSLs were identified from three strains of Skeletonema, which might have important biological functions. It could also provide a reliable tool for chemotaxonomy of microalgae. Copyright © 2013 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

A method has been established by which to determine aldoses and ketoses in plant material simultaneously. Monosaccharides were extracted by sonication with 80% ethanol and sugar oximes formed by treatment of the resultant extract with hydroxylamine and pyridine at 90C. After reaction, one aliquot of the product was derivatised with acetic anhydride at 90C, whilst a second aliquot was silylated with HMDS and TMCS at 80C. Both reaction mixtures were analysed by GC-MS in the SIM mode. Quantivation was linear within the range 1-4 mg/mL and the detection limit for monosaccharides was 5-25 ng/mL. The absolute recoveries were between 73.0 and 90.2% and the RSDs were 3.1-10.0%. This method was applied to analyse the free monosaccharides in Lyceum barbarum L.; eight monosaccharides were present in amounts between 0.26 and 368.65 mg/mg. Copyright © 2006 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]