BACKGROUND: Flax (Linum usitatissimum L.) is grown for fiber and seed in many countries. Flax cultivars differ in the oil composition and, depending on the ratio of fatty acids, are used in pharmaceutical, food, or paint industries. It is known that genes of SAD (stearoyl-ACP desaturase) and FAD (fatty acid desaturase) families play a key role in the synthesis of fatty acids, and some alleles of these genes are associated with a certain composition of flax oil. However, data on genetic polymorphism of these genes are still insufficient. RESULTS: On the basis of the collection of the Institute for Flax (Torzhok, Russia), we formed a representative set of 84 cultivars and lines reflecting the diversity of fatty acid composition of flax oil. An approach for the determination of full-length sequences of SAD1, SAD2, FAD2A, FAD2B, FAD3A, and FAD3B genes using the Illumina platform was developed and deep sequencing of the 6 genes in 84 flax samples was performed on MiSeq. The obtained high coverage (about 400x on average) enabled accurate assessment of polymorphisms in SAD1, SAD2, FAD2A, FAD2B, FAD3A, and FAD3B genes and evaluation of cultivar/line heterogeneity. The highest level of genetic diversity was observed for FAD3A and FAD3B genes - 91 and 62 polymorphisms respectively. Correlation analysis revealed associations between particular variants in SAD and FAD genes and predominantly those fatty acids whose conversion they catalyze: SAD - stearic and oleic acids, FAD2 - oleic and linoleic acids, FAD3 - linoleic and linolenic acids. All except one low-linolenic flax cultivars/lines contained both the substitution of tryptophan to stop codon in the FAD3A gene and histidine to tyrosine substitution in the FAD3B gene, while samples with only one of these polymorphisms had medium content of linolenic acid and cultivars/lines without them were high-linolenic. CONCLUSIONS: Genetic polymorphism of SAD and FAD genes was evaluated in the collection of flax cultivars and lines with diverse oil composition, and associations between particular polymorphisms and the ratio of fatty acids were revealed. The achieved results are the basis for the development of marker-assisted selection and DNA-based certification of flax cultivars.
- MeSH
- desaturasy mastných kyselin genetika MeSH
- DNA rostlinná MeSH
- genetická heterogenita MeSH
- genetická variace * MeSH
- kyselina alfa-linolenová metabolismus MeSH
- len enzymologie genetika metabolismus MeSH
- mastné kyseliny metabolismus MeSH
- oxygenasy se smíšenou funkcí genetika metabolismus MeSH
- rostlinné geny MeSH
- sekvenční analýza DNA MeSH
- substituce aminokyselin MeSH
- Publikační typ
- časopisecké články MeSH
Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a major cause of foodborne gastrointestinal illness. The adhesion of EHEC to host tissues is the first step enabling bacterial colonization. Adhesins such as fimbriae and flagella mediate this process. Here, we studied the interaction of the bacterial flagellum with the host cell's plasma membrane using giant unilamellar vesicles (GUVs) as a biologically relevant model. Cultured cell lines contain many different molecular components, including proteins and glycoproteins. In contrast, with GUVs, we can characterize the bacterial mode of interaction solely with a defined lipid part of the cell membrane. Bacterial adhesion on GUVs was dependent on the presence of the flagellar filament and its motility. By testing different phospholipid head groups, the nature of the fatty acid chains, or the liposome curvature, we found that lipid packing is a key parameter to enable bacterial adhesion. Using HT-29 cells grown in the presence of polyunsaturated fatty acid (α-linolenic acid) or saturated fatty acid (palmitic acid), we found that α-linolenic acid reduced adhesion of wild-type EHEC but not of a nonflagellated mutant. Finally, our results reveal that the presence of flagella is advantageous for the bacteria to bind to lipid rafts. We speculate that polyunsaturated fatty acids prevent flagellar adhesion on membrane bilayers and play a clear role for optimal host colonization. Flagellum-mediated adhesion to plasma membranes has broad implications for host-pathogen interactions.IMPORTANCE Bacterial adhesion is a crucial step to allow bacteria to colonize their hosts, invade tissues, and form biofilm. Enterohemorrhagic Escherichia coli O157:H7 is a human pathogen and the causative agent of diarrhea and hemorrhagic colitis. Here, we use biomimetic membrane models and cell lines to decipher the impact of lipid content of the plasma membrane on enterohemorrhagic E. coli flagellum-mediated adhesion. Our findings provide evidence that polyunsaturated fatty acid (α-linolenic acid) inhibits E. coli flagellar adhesion to the plasma membrane in a mechanism separate from its antimicrobial and anti-inflammatory functions. In addition, we confirm that cholesterol-enriched lipid microdomains, often called lipid rafts, are important in bacterial adhesion. These findings demonstrate that plasma membrane adhesion via bacterial flagella play a significant role for an important human pathogen. This mechanism represents a promising target for the development of novel antiadhesion therapies.
- MeSH
- bakteriální adheze * MeSH
- buněčná membrána chemie MeSH
- buněčné linie MeSH
- buňky HT-29 MeSH
- epitelové buňky mikrobiologie MeSH
- Escherichia coli O157 fyziologie MeSH
- flagella metabolismus MeSH
- fosfolipidy analýza MeSH
- interakce hostitele a patogenu * MeSH
- kyselina alfa-linolenová analýza MeSH
- kyselina palmitová analýza MeSH
- lidé MeSH
- membránové mikrodomény chemie MeSH
- unilamelární lipozómy chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A terrestrial green microalga was isolated at Ås, in Akershus County, Norway. The strain corresponded to a coccoid chlorophyte. Morphological characteristics by light and electron microscopy, in conjunction with DNA amplification and sequencing of the 18 s rDNA gene and ITS sequences, were used to identify the microalgae. The characteristics agree with those of the genus Coelastrella defined by Chodat, and formed a sister group with the recently described C. thermophila var. globulina. Coelastrella is a relatively small numbered genus that has not been observed in continental Norway before; there are no previous cultures available in collections of Norwegian strains. Gas chromatography analyses of the FAME-derivatives showed a high percentage of polyunsaturated fatty acids (44-45%) especially linolenic acid (C18:3n3; 30-34%). After the stationary phase, the cultures were able to accumulate several carotenoids as neoxanthin, pheophytin a, astaxanthin, canthaxanthin, lutein, and violaxanthin. Due to the scarcity of visual characters suitable for diagnostic purposes and the lack of DNA sequence information, there is a high possibility that species of this genus have been neglected in local environmental studies, even though it showed interesting properties for algal biotechnology.
- MeSH
- biologické pigmenty analýza MeSH
- biotechnologie MeSH
- Chlorophyta klasifikace cytologie genetika MeSH
- druhová specificita MeSH
- feofytiny analýza MeSH
- fylogeneze * MeSH
- karotenoidy analýza MeSH
- kyselina alfa-linolenová analýza MeSH
- mastné kyseliny analýza MeSH
- mikrořasy klasifikace cytologie genetika izolace a purifikace MeSH
- ribozomální DNA MeSH
- RNA ribozomální 18S genetika MeSH
- xanthofyly MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Norsko MeSH
: Jasmonic acid (JA) and its related derivatives are ubiquitously occurring compounds of land plants acting in numerous stress responses and development. Recent studies on evolution of JA and other oxylipins indicated conserved biosynthesis. JA formation is initiated by oxygenation of α-linolenic acid (α-LeA, 18:3) or 16:3 fatty acid of chloroplast membranes leading to 12-oxo-phytodienoic acid (OPDA) as intermediate compound, but in Marchantiapolymorpha and Physcomitrellapatens, OPDA and some of its derivatives are final products active in a conserved signaling pathway. JA formation and its metabolic conversion take place in chloroplasts, peroxisomes and cytosol, respectively. Metabolites of JA are formed in 12 different pathways leading to active, inactive and partially active compounds. The isoleucine conjugate of JA (JA-Ile) is the ligand of the receptor component COI1 in vascular plants, whereas in the bryophyte M. polymorpha COI1 perceives an OPDA derivative indicating its functionally conserved activity. JA-induced gene expressions in the numerous biotic and abiotic stress responses and development are initiated in a well-studied complex regulation by homeostasis of transcription factors functioning as repressors and activators.
- MeSH
- chloroplasty metabolismus MeSH
- cyklopentany metabolismus MeSH
- druhová specificita MeSH
- kyselina alfa-linolenová metabolismus MeSH
- Marchantia metabolismus MeSH
- mastné kyseliny metabolismus MeSH
- mechy metabolismus MeSH
- metabolické sítě a dráhy MeSH
- nenasycené mastné kyseliny metabolismus MeSH
- oxylipiny metabolismus MeSH
- peroxizomy metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Activation of calmodulin dependent protein kinase (CaMK)II by exercise is beneficial in controlling membrane lipids associated with type 2 diabetes and obesity. Regulation of lipid metabolism is crucial in the improvement of type 2 diabetes and obesity associated symptoms. The role of CaMKII in membrane associated lipid metabolism was the focus of this study. Five to six weeks old male Wistar rats were used in this study. GC×GC-TOFMS technique was used to determine the levels of polyunsaturated fatty acids (linoleic acid, arachidonic acid and 11,14-eicosadienoic acid). Carnitine palmitoyltransferase (Cpt-1) and acetyl-CoA carboxylase (Acc-1) genes expression were assessed using quantitative real time PCR (qPCR). From the results, CaMKII activation by exercise increased the levels of arachidonic acid and 11,14-eicosadienoic acid while a decrease in the level of linolenic acid was observed in the skeletal muscle. The results indicated that exercise-induced CaMKII activation increased CPT-1 expression and decreased ACC-1 expression in rat skeletal muscle. All the observed increases with activation of CaMKII by exercise were aborted when KN93, an inhibitor of CaMKII was injected in exercising rats. This study demonstrated that CaMKII activation by exercise regulated lipid metabolism. This study suggests that CaMKII can be a vital target of therapeutic approach in the management of diseases such as type 2 diabetes and obesity that have increased to epidemic proportions recently.
- MeSH
- acetyl-CoA-karboxylasa genetika metabolismus MeSH
- aktivace enzymů MeSH
- fosforylace MeSH
- karnitin-O-palmitoyltransferasa genetika metabolismus MeSH
- kondiční příprava zvířat * MeSH
- kosterní svaly enzymologie MeSH
- kyselina alfa-linolenová metabolismus MeSH
- kyselina arachidonová metabolismus MeSH
- mastné kyseliny metabolismus MeSH
- messenger RNA genetika metabolismus MeSH
- potkani Wistar MeSH
- proteinkinasa závislá na vápníku a kalmodulinu typ 2 metabolismus MeSH
- svalová kontrakce * MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Klíčová slova
- Linum usitatissimum L., len setý, Linum crepitans (Boenn.) Dumort., Linum humile Mill., Flachs (něm.), Flax (angl.), Lin (franc.),
- MeSH
- fytoterapie * MeSH
- kyselina alfa-linolenová * terapeutické užití MeSH
- kyselina linolová * terapeutické užití MeSH
- laktace MeSH
- léčivé rostliny * MeSH
- lékové interakce * MeSH
- len * chemie MeSH
- lidé MeSH
- lignany * terapeutické užití MeSH
- semena rostlinná * MeSH
- těhotenství MeSH
- Check Tag
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
The objective of the study is to standardize GC determination of polyunsaturated fatty acids in animal tissues by lipid extraction with a hexane/propan-2-ol mixture and subsequent derivatization with CH3ONa/BF3. The limits of detection and recoveries for 18:2n-6, 20:4n-6, 18:3n-3, 20:5n-3, 22:5n-3 and 22:6n-3 fatty acids was in the range 0.32–0.42 mg/100 g, and 83–105 %, respectively. The content of physiologically important icosapentaenoic acid (EPA) in chicken breast meat, thigh meat, quail eggs and cricket nymphs was 3, 10, 51 and 21 mg/100 g of fresh matter (P < 0.05). The EPA contents in livers of laboratory rats fed diets supplemented with palm oil, safflower oil and fish oil was 4, 3 and 342 mg/100 g, respectively (P < 0.05).
- MeSH
- cholesterol metabolismus MeSH
- chromatografie plynová * metody přístrojové vybavení využití MeSH
- experimenty na zvířatech MeSH
- kardiovaskulární nemoci prevence a kontrola MeSH
- kyselina alfa-linolenová analýza MeSH
- kyselina eikosapentaenová analýza MeSH
- kyseliny dokosahexaenové analýza MeSH
- lidé MeSH
- lipidy analýza MeSH
- nenasycené mastné kyseliny * analýza MeSH
- tkáňové extrakty analýza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Total lipid contents of green (Chlorella pyrenoidosa, C), red (Porphyra tenera, N; Palmaria palmata, D), and brown (Laminaria japonica, K; Eisenia bicyclis, A; Undaria pinnatifida, W, WI; Hizikia fusiformis, H) commercial edible algal and cyanobacterial (Spirulina platensis, S) products, and autotrophically cultivated samples of the green microalga Chlorella kessleri (CK) and the cyanobacterium Spirulina platensis (SP) were determined using a solvent mixture of methanol/chloroform/water (1:2:1, v/v/v, solvent I) and n-hexane (solvent II). Total lipid contents ranged from 0.64% (II) to 18.02% (I) by dry weight and the highest total lipid content was observed in the autotrophically cultivated cyanobacterium Spirulina platensis. Solvent mixture I was found to be more effective than solvent II. Fatty acids were determined by gas chromatography of their methyl esters (% of total FAMEs). Generally, the predominant fatty acids (all results for extractions with solvent mixture I) were saturated palmitic acid (C16:0; 24.64%-65.49%), monounsaturated oleic acid (C18:1(n-9); 2.79%-26.45%), polyunsaturated linoleic acid (C18:2(n-6); 0.71%-36.38%), α-linolenic acid (C18:3(n-3); 0.00%-21.29%), γ-linolenic acid (C18:3(n-6); 1.94%-17.36%), and arachidonic acid (C20:4(n-6); 0.00%-15.37%). The highest content of ω-3 fatty acids (21.29%) was determined in Chlorella pyrenoidosa using solvent I, while conversely, the highest content of ω-6 fatty acids (41.42%) was observed in Chlorella kessleri using the same solvent.
- MeSH
- analýza potravin MeSH
- Chlorophyta chemie MeSH
- kyselina alfa-linolenová chemie MeSH
- kyselina linolová chemie MeSH
- kyselina palmitová chemie MeSH
- kyseliny mastné omega-3 chemie MeSH
- lipidy chemie izolace a purifikace MeSH
- mořské řasy chemie MeSH
- rozpouštědla chemie MeSH
- sinice chemie MeSH
- sladká voda MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
