scholarly journals Fatty Acid and Associated Gene Expression Analyses of Three Tree Peony Species Reveal Key Genes for α-Linolenic Acid Synthesis in Seeds

2018 ◽  
Vol 9 ◽  
Author(s):  
Qing-Yu Zhang ◽  
Rui Yu ◽  
Li-Hang Xie ◽  
Md Mahbubur Rahman ◽  
Aruna Kilaru ◽  
...  
2018 ◽  
Vol 20 (1) ◽  
pp. 65 ◽  
Author(s):  
Qingyu Zhang ◽  
Rui Yu ◽  
Daoyang Sun ◽  
Md Rahman ◽  
Lihang Xie ◽  
...  

Tree peony (Paeonia section Moutan DC.) species are woody oil crops with high unsaturated fatty acid content, including α-linolenic acid (ALA/18:3; >40% of the total fatty acid). Comparative transcriptome analyses were carried out to uncover the underlying mechanisms responsible for high and low ALA content in the developing seeds of P. rockii and P. lutea, respectively. Expression analysis of acyl lipid metabolism genes revealed upregulation of select genes involved in plastidial fatty acid synthesis, acyl editing, desaturation, and triacylglycerol assembly in seeds of P. rockii relative to P. lutea. Also, in association with ALA content in seeds, transcript levels for fatty acid desaturases (SAD, FAD2, and FAD3), which encode enzymes necessary for polyunsaturated fatty acid synthesis, were higher in P. rockii compared to P. lutea. Furthermore, the overexpression of PrFAD2 and PrFAD3 in Arabidopsis increased linoleic and ALA content, respectively, and modulated the final ratio 18:2/18:3 in the seed oil. In conclusion, we identified the key steps and validated the necessary desaturases that contribute to efficient ALA synthesis in a woody oil crop. Together, these results will aid to increase essential fatty acid content in seeds of tree peonies and other crops of agronomic interest.


2012 ◽  
Vol 78 (6) ◽  
pp. 1865-1875 ◽  
Author(s):  
Anna E. Nikitkova ◽  
Elaine M. Haase ◽  
M. Margaret Vickerman ◽  
Steven R. Gill ◽  
Frank A. Scannapieco

ABSTRACTStreptococcus gordonii, an important primary colonizer of dental plaque biofilm, specifically binds to salivary amylase via the surface-associated amylase-binding protein A (AbpA). We hypothesized that a function of amylase binding toS. gordoniimay be to modulate the expression of chromosomal genes, which could influence bacterial survival and persistence in the oral cavity. Gene expression profiling by microarray analysis was performed to detect genes inS. gordoniistrain CH1 that were differentially expressed in response to the binding of purified human salivary amylase versus exposure to purified heat-denatured amylase. Selected genes found to be differentially expressed were validated by quantitative reverse transcription-PCR (qRT-PCR). Five genes from the fatty acid synthesis (FAS) cluster were highly (10- to 35-fold) upregulated inS. gordoniiCH1 cells treated with native amylase relative to those treated with denatured amylase. AnabpA-deficient strain ofS. gordoniiexposed to amylase failed to show a response in FAS gene expression similar to that observed in the parental strain. Predicted phenotypic effects of amylase binding toS. gordoniistrain CH1 (associated with increased expression of FAS genes, leading to changes in fatty acid synthesis) were noted; these included increased bacterial growth, survival at low pH, and resistance to triclosan. These changes were not observed in the amylase-exposedabpA-deficient strain, suggesting a role for AbpA in the amylase-induced phenotype. These results provide evidence that the binding of salivary amylase elicits a differential gene response inS. gordonii, resulting in a phenotypic adjustment that is potentially advantageous for bacterial survival in the oral environment.


2021 ◽  
Vol 27 (Supplement_1) ◽  
pp. S9-S10
Author(s):  
Brooklyn McGrew ◽  
Aman Shrivastava ◽  
Philip Fernandes ◽  
Lubaina Ehsan ◽  
Yash Sharma ◽  
...  

Abstract Background Candidate markers for Crohn’s Disease (CD) may be identified via gene expression-based construction of metabolic networks (MN). These can computationally describe gene-protein-reaction associations for entire tissues and also predict the flux of reactions (rate of turnover of specific molecules via a metabolic pathway). Recon3D is the most comprehensive human MN to date. We used publicly available CD transcriptomic data along with Recon3D to identify metabolites as potential diagnostic and prognostic biomarkers. Methods Terminal ileal gene expression profiles (36,372 genes; 218 CD. 42 controls) from the RISK cohort (Risk Stratification and Identification of Immunogenetic and Microbial Markers of Rapid Disease Progression in Children with Crohn’s Disease) and their transcriptomic abundances were used. Recon3D was pruned to only include RISK dataset transcripts which determined metabolic reaction linkage with transcriptionally active genes. Flux balance analysis (FBA) was then run using RiPTiDe with context specific transcriptomic data to further constrain genes (Figure 1). RiPTiDe was independently run on transcriptomic data from both CD and controls. From the pruned and constricted MN obtained, reactions were extracted for further analysis. Results After applying the necessary constraints to modify Recon3D, 527 CD and 537 control reactions were obtained. Reaction comparison with a publicly available list of healthy small intestinal epithelial reactions (n=1282) showed an overlap of 80 CD and 84 control reactions. These were then further grouped based on their metabolic pathways. RiPTiDe identified context specific metabolic pathway activity without supervision and the percentage of forward, backward, and balanced reactions for each metabolic pathway (Figure 2). The metabolite concentrations in the small intestine was altered among CD patients. Notably, the citric acid cycle and malate-aspartate shuttle were affected, highlighting changes in mitochondrial metabolic pathways. This is illustrated by changes in the number of reactions at equilibrium between CD and control. Conclusions The results are relevant as cytosolic acetyl-CoA is needed for fatty acid synthesis and is obtained by removing citrate from the citric acid cycle. An intermediate removal from the cycle has significant cataplerotic effects. The malate-aspartate shuttle also allows electrons to move across the impermeable membrane in the mitochondria (fatty acid synthesis location). These findings are reported by previously published studies where gene expression for fatty acid synthesis is altered in CD patients along with mitochondrial metabolic pathway changes, resulting in altered cell homeostasis. In-depth analysis is currently underway with our work supporting the utility of potential metabolic biomarkers for CD diagnosis, management and improved care.


Nutrients ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 723 ◽  
Author(s):  
Eric Ponnampalam ◽  
Vahid Vahedi ◽  
Khageswor Giri ◽  
Paul Lewandowski ◽  
Joe Jacobs ◽  
...  

This study investigated the effect of dietary manipulations on muscle fatty acid composition, the activities and relative mRNA expressions of antioxidant enzymes and the relationship between muscle enzyme activity or mRNA expression and alpha linolenic acid (ALA) concentration in sheep. Eighty-four lambs blocked on liveweight were randomly allocated to four dietary treatments, lucerne pasture (Lucerne), annual ryegrass pasture (Ryegrass), feedlot pellets (Feedlot) or annual ryegrass plus feedlot pellets (RyeFeedlot). After six weeks of feeding, lambs were slaughtered and within 30 min post-mortem, samples collected from the longissimus lumborum (LL) muscle for RNA isolation and measurement of antioxidant enzyme activities. At 24 h post-mortem, LL samples were collected for determination of fatty acid concentrations. Feedlot treatment decreased ALA, eicosapentaenoic (EPA), docosapentaenoic (DPA) and docosahexaenoic (DHA) concentrations compared with other treatments and increased linoleic acid (LA) and arachidonic acid (AA) compared with Lucerne and Ryegrass (p < 0.001). The activity of Glutathione peroxidase (GPX1, p < 0.001) and Superoxide dismutase (SOD2, p < 0.001) enzymes in the muscle increased with Lucerne compared to other treatments. Lucerne increased muscle gpx1 mRNA expression by 1.74-fold (p = 0.01) and 1.68-fold (p = 0.05) compared with Feedlot and other diets, respectively. The GPX1 (r2 = 0.319, p = 0.002) and SOD2 (r2 = 0.244, p = 0.009) enzyme activities were positively related to ALA. There was a positive linear relationship between muscle gpx1 (r2 = 0.102, p = 0.017) or sod2 (r2 = 0.049, p = 0.09) mRNA expressions and ALA concentration. This study demonstrates that diet can affect concentrations of ALA and other fatty acids as well as change activities and gene expression of antioxidant enzymes in muscle. Increased antioxidant activity may, in turn, have beneficial effects on the performance, health and wellbeing of animals and humans.


Animals ◽  
2019 ◽  
Vol 9 (7) ◽  
pp. 467 ◽  
Author(s):  
Ahmed A. Saleh ◽  
Abeer A. Kirrella ◽  
Safaa E. Abdo ◽  
Mahmoud M. Mousa ◽  
Nemat A. Badwi ◽  
...  

The present study was conducted to examine that impact of dietary xylanase (Xyl) and arabinofuranosidase (Abf) supplementation on the performance, protein and fat digestibility, the lipid peroxidation, the plasma biochemical traits, and the immune response of broilers. A total of 480, un-sexed, and one-day-old broilers (Ross 308) were randomly divided into three treatments with eight replicates, where chicks in the first treatment were fed basal diets and served as the control, chicks in the second treatment were fed diets formulated with reductions of 90 kcal/kg, and chicks in the third treatment were fed the same formulated diets used in the second group as well as the Xyl and Abf combination (Rovabio® Advance). Feed intake was decreased by the low energy diet, leading to an enhancement in feed efficiency enzyme supplementation in the low energy diet (p < 0.015). Both protein and fat digestibility were improved (p < 0.047) due to enzyme supplementation. Moreover, enzyme supplementation increased muscle total lipids content and decreased muscle thiobarbituric acid retroactive substance content. Furthermore, diets supplemented with Xyl and Abf exhibited an increase in antibody titers against the Newcastle disease virus (p < 0.026). In addition, enzyme supplementation increased gene expression related to growth and gene expression related to fatty acid synthesis. It could be concluded that dietary Xyl and Abf supplementation had beneficial impacts on growth, nutrient digestibility, lipid peroxidation, immune response, and gene expressions related to growth and fatty acid synthesis in broiler chickens fed low-energy diets.


2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 1270-1270
Author(s):  
Samantha Pauls ◽  
Christopher Pascoe ◽  
Lisa Rodway ◽  
Carla Taylor ◽  
Harold Aukema ◽  
...  

Abstract Objectives Docosahexaenoic acid (DHA) can be obtained directly from the diet or produced by elongation and desaturation of α-linolenic acid (ALA). Both are proposed to reduce inflammation associated with obesity, however, fewer studies have investigated ALA. The objective of this study was to evaluate the gene expression changes in monocytes induced by each fatty acid and to compare the predicted functional outcomes. Methods RNA was extracted from THP-1 monocytes treated with ALA, DHA or vehicle for 48 h and then transcriptomics profiles were assessed by microarray. Multiple tools were used for data interpretation, including fold change analysis, Principal Component Analysis (PCA), Variable Importance Projection (VIP), Ingenuity Pathway Analysis (IPA) and Network Analyst. Results We found that the ALA and DHA treatments produced distinct profiles with many individual genes making small contributions to the separation between groups. Relative to vehicle treatment, many downregulated targets were similarly affected by both ALA and DHA. Several of these downregulated genes are involved in cholesterol synthesis and are regulated by miR-335–5p, a microRNA upregulated by both treatments. Consistently, IPA predicted similar pathways and functions are decreased by ALA and DHA, most notably cholesterol biosynthesis. In contrast, ALA and DHA upregulated unique gene sets and in agreement IPA predicted each treatment would activate distinct pathways and functions. ALA was strongly and uniquely predicted to increase infection responses while only DHA was predicted to increase oxidative phosphorylation. Finally, analysis of the protein-protein interaction network involving the genes modified by each fatty acid treatment allowed us to predict the most functionally important gene targets, which will be tested in future studies. Conclusions These analyses have revealed both unique and overlapping effects of ALA and DHA on the monocyte gene expression profile, providing further evidence that they have distinct bioactivities. Many novel predictions were made and these will form the basis for future studies investigating the effects of ALA and DHA on human physiology. Funding Sources Natural Sciences and Engineering Research Council of Canada; Canadian Institutes of Health Research.


2012 ◽  
Vol 302 (2) ◽  
pp. E209-E217 ◽  
Author(s):  
Kristian K. Jensen ◽  
Stephen F. Previs ◽  
Lei Zhu ◽  
Kithsiri Herath ◽  
Sheng-Ping Wang ◽  
...  

The liver is a crossroad for metabolism of lipid and carbohydrates, with acetyl-CoA serving as an important metabolic intermediate and a precursor for fatty acid and cholesterol biosynthesis pathways. A better understanding of the regulation of these pathways requires an experimental approach that provides both quantitative metabolic flux measurements and mechanistic insight. Under conditions of high carbohydrate availability, excess carbon is converted into free fatty acids and triglyceride for storage, but it is not clear how excessive carbohydrate availability affects cholesterol biosynthesis. To address this, C57BL/6J mice were fed either a low-fat, high-carbohydrate diet or a high-fat, carbohydrate-free diet. At the end of the dietary intervention, the two groups received 2H2O to trace de novo fatty acid and cholesterol synthesis, and livers were collected for gene expression analysis. Expression of lipid and glucose metabolism genes was determined using a custom-designed pathway focused PCR-based gene expression array. The expression analysis showed downregulation of cholesterol biosynthesis genes and upregulation of fatty acid synthesis genes in mice receiving the high-carbohydrate diet compared with the carbohydrate-free diet. In support of these findings, 2H2O tracer data showed that fatty acid synthesis was increased 10-fold and cholesterol synthesis was reduced by 1.6-fold in mice fed the respective diets. In conclusion, by applying gene expression analysis and tracer methodology, we show that fatty acid and cholesterol synthesis are differentially regulated when the carbohydrate intake in mice is altered.


2014 ◽  
Vol 54 (9) ◽  
pp. 1436 ◽  
Author(s):  
B. P. Dalrymple ◽  
B. Guo ◽  
G. H. Zhou ◽  
W. Zhang

Intramuscular fat content (IMF%) in cattle influences the value of individual animals, especially for higher marbling markets. IMF is triacylglyceride (TAG) in lipid droplets in the intramuscular adipocytes. However, there are many different pathways from feed intake to the final common process of TAG synthesis and storage as IMF. To evaluate the relative importance of different pathways we compared changes in the expression of genes encoding proteins involved in the TAG and fatty acid (FA) synthesis pathways in the longissimus muscle of Piedmontese × Hereford (P×H) and Wagyu × Hereford (W×H) crosses. Based on these changes we have estimated the relative contributions of FA synthesised de novo in the intramuscular adipocyte and the uptake of circulating FA (both free and from TAG), from the diet or synthesised de novo in other tissues, to TAG deposition as IMF. We have analysed the impact of different developmental times and different diets on these processes. Increased de novo FA synthesis in intramuscular adipocytes appeared to contribute more than increased FA uptake from circulation to the additional TAG deposition in W×H compared with P×H cattle between 12 and 25 months (forage diet). Changing diet from forage to concentrate appeared to increase the importance of FA uptake from circulation relative to de novo FA synthesis for TAG synthesis in intramuscular adipocytes. These results are consistent with the literature based on analysis of lipid composition. Gene expression appears to provide a simple assay for identification of the source of FA for the deposition of IMF.


2021 ◽  
Author(s):  
Sumie Eto ◽  
Rumie Matsumura ◽  
Mai Fujimi ◽  
Yasuhiro Shimane ◽  
Samuel Berhanu ◽  
...  

Phospholipid synthesis is a fundamental process that promotes cell propagation and, presently, is the most challenging issue in artificial cell research aimed at reconstituting living cells from biomolecules. Here, we constructed a cell-free phospholipid synthesis system that combines in vitro fatty acid synthesis and a cell-free gene expression system that synthesizes acyltransferases for phospholipid synthesis. Fatty acids were synthesized from acetyl-CoA and malonyl-CoA, then continuously converted into phosphatidic acids by the cell-free synthesized acyltransferases. Because the system can avoid the accumulation of synthetic intermediates that suppress the reaction, the yield of phospholipid has significantly improved from previous schemes (up to 400 μM). Additionally, by adding enzymes for recycling CoA, we synthesized phosphatidic acids from acetic acid and bicarbonate as carbon sources. The constructed system is available to express the genes from pathogenic bacteria and to analyze the synthesized phospholipids. By encapsulating our system inside giant vesicles, it would be possible to construct the artificial cells in which the membrane grows and divides sustainably.


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