Omega-3 Fatty Acid Desaturase Research Articles

Biosynthetic deficiency of docosahexaenoic acid causes nonalcoholic fatty liver disease and ferroptosis-mediated hepatocyte injury

Exogenous omega-3 fatty acids, particularly docosahexaenoic acid (DHA), have shown to exert beneficial effects on nonalcoholic fatty liver disease (NAFLD), which is characterized by the excessive accumulation of lipids and chronic injury in the liver. However, the effect of endogenous DHA biosynthesis on the lipid homeostasis of liver is poorly understood. In this study, we used a DHA biosynthesis-deficient zebrafish model, elovl2 mutant, to explore the effect of endogenously biosynthesized DHA on hepatic lipid homeostasis. We found the pathways of lipogenesis and lipid uptake were strongly activated, while the pathways of lipid oxidation and lipid transport were inhibited in the liver of elovl2 mutants, leading to lipid droplet accumulation in the mutant hepatocytes and NAFLD. Furthermore, the elovl2 mutant hepatocytes exhibited disrupted mitochondrial structure and function, activated endoplasmic reticulum (ER) stress, and hepatic injury. We further unveiled that the hepatic cell death and injury was mainly mediated by ferroptosis, rather than apoptosis, in elovl2 mutants. Elevating DHA content in elovl2 mutants, either through introduction of an omega-3 desaturase (fat1) transgene or feeding with a DHA-rich diet, could strongly alleviate NAFLD features and ferroptosis-mediated hepatic injury. Together, our study elucidates the essential role of endogenous DHA biosynthesis in maintaining hepatic lipid homeostasis and liver health, highlighting that DHA deficiency can lead to NAFLD and ferroptosis-mediated hepatic injury.

Endogenous biosynthesis of docosahexaenoic acid (DHA) regulates fish oocyte maturation by promoting pregnenolone production.

Omega-3 polyunsaturated fatty acids (n-3 PUFAs), particularly docosahexaenoic acid (22:6n-3, DHA), play crucial roles in the reproductive health of vertebrates, including humans. Nevertheless, the underlying mechanism related to this phenomenon remains largely unknown. In this study, we employed two zebrafish genetic models, i.e., elovl2 -/- mutant as an endogenous DHA-deficient model and fat1 (omega-3 desaturase encoding gene) transgenic zebrafish as an endogenous DHA-rich model, to investigate the effects of DHA on oocyte maturation and quality. Results show that the elovl2 -/- mutants had much lower fecundity and poorer oocyte quality than the wild-type controls, while the fat1 zebrafish had higher fecundity and better oocyte quality than wild-type controls. DHA deficiency in elovl2 -/- embryos led to defects in egg activation, poor microtubule stability, and reduced pregnenolone levels. Further study revealed that DHA promoted pregnenolone synthesis by enhancing transcription of cyp11a1, which encodes the cholesterol side-chain cleavage enzyme, thereby stabilizing microtubule assembly during oogenesis. In turn, the hypothalamic-pituitary-gonadal axis was enhanced by DHA. In conclusion, using two unique genetic models, our findings demonstrate that endogenously synthesized DHA promotes oocyte maturation and quality by promoting pregnenolone production via transcriptional regulation of cyp11a1.

Integrated lipidomic and transcriptomic analysis reveals diacylglycerol accumulation in olive of Longnan (China).

Olive (Olea europaea L.) oil accumulate more diacylglycerols (DAG) than mostly vegetable oils. Unsaturated fatty acids-enriched DAG consumption enhanced wellness in subjects. However, the mechanism of DAG accumulation is not yet fully understood. In this study, gene network of DAG accumulation and fatty acid composition in the two olive mesocarps ("Chenggu 32" (CG) and "Koroneiki" (QJ)) were investigated by integrating lipidome and transcriptome techniques. A total of 1,408 lipid molecules were identified by lipidomic analysis in olive mesocarp, of which DAG (DAG36:3, DAG36:4 and DAG36:5) showed higher content, and triacylglycerols (TAG54:3, TAG54:4) exhibited opposite trend in CG. Specifically, DAG was rich in polyunsaturated fatty acids (especially C18:2) at the sn-2 position, which was inconsistent with TAG at the same positions (Primarily C18:1). Transcriptomic analysis revealed that phospholipase C (NPC, EC 3.1.4.3) were up-regulated relative to QJ, whereas diacylglycerol kinase (ATP) (DGK, EC 2.7.1.107), diacylglycerol acyltransferase (DGAT, EC 2.3.1.20), and phospholipid: diacylglycerol acyltransferase (PDAT, EC 2.3.1.158) were down-regulated. We speculated that the non-acyl coenzyme A pathway played a significant role in DAG biosynthesis. Additionally, fatty acyl-ACP thioesterase B (FATB, EC 3.1.2.14), stearoyl [acyl-carrier-protein] 9-desaturase (SAD, EC 1.14.19.2) and omega-6 fatty acid desaturase (FAD2, EC 1.14.19.6) were highly expressed in CG and may be involved in regulating fatty acid composition. Meanwhile, phospholipase A1 (LCAT, EC 3.1.1.32) involved in the acyl editing reaction facilitated PUFA linkage at the sn-2 position of DAG. Our findings provide novel insights to increase the DAG content, improve the fatty acid composition of olive oil, and identify candidate genes for the production of DAG-rich oils.

Comparative analysis of omega-3 fatty acid desaturase FAD7A-1 chloroplastic gene in castor

Omega-3 Polyunsaturated fatty acids (PUFAs) are characterized by the presence of a double bond, three atoms away from the terminal methyl group in their chemical structure. These fatty acids play a crucial role in promoting health benefits. Castor, known for its applications in traditional medicine and as a lubricant, has been historically employed for its pain-relieving properties and enhancement of muscle activity. In the present study the omega-3 fatty acid desaturase FAD7A-1 chloroplastic (P48619, FAD3C_RICCO) gene sequence of castor was obtained from the NCBI data base was analyzed for its diversity, amino acid composition and phylogenetic relatedness with other species harboring the gene. The phylogenetic tree of 13 aligned sequences clustered into two clusters and the amino acids varied between the species evaluated.

Multi-omics integration to explore the molecular insight into the volatile organic compounds in watermelon

A range of volatile organic compounds played an important role in the formation of watermelon fruit aroma, while due to the low content and difficulty in detection, it is often neglected in watermelon breeding programs, resulting in a decline in fruit flavor. VOCs in the flesh of 194 watermelon accessions and seven cultivars at four developmental stages were determined by SPME-GC–MS. Ten metabolites with significant differences in the natural population and positive accumulation during fruit development are considered to be the key metabolite related to watermelon fruit aroma. And the link between metabolite and, flesh color and sugar content by correlation analysis was established. The results of the genome-wide association study showed that (5E)-6,10-dimethylundeca-5,9-dien-2-one, and 1-(4-methylphenyl) ethanone were colocalized with watermelon flesh color on chromosome 4, which may be regulated by LCYB and CCD. (E)-4-(2,6,6-trimethylcyclohexen-1-yl)but-3-en-2-one is the VOC produced by the cleavage of carotenoids, which has a positive correlation with the sugar content of the fruit, and the candidate gene Cla97C05G092490 on chromosome 5 may interact with PSY to influence the accumulation of this metabolite. In addition, Cla97C02G049790 (enol reductase), Cla97C03G051490 (omega-3 fatty acid desaturase gene), LOX, and ADH may play important roles in the synthesis of fatty acids and their derived VOCs. Taken together, our findings provide molecular insights into the accumulation and natural variation of VOCs in watermelon, and give data support for breeding watermelon cultivars with better flavor.

Modulation of GmFAD3 expression alters abiotic stress responses in soybean.

This study focused on enhancing resilience of soybean crops to drought and salinity stresses by overexpression of GmFAD3A gene, which plays an important role in modulating membrane fluidity and ultimately influence plants response to various abiotic stresses. Fatty acid desaturases (FADs) are a class of enzymes that mediate desaturation of fatty acids by introducing double bonds. They play an important role in modulating membrane fluidity in response to various abiotic stresses. However, a comprehensive analysis of GmFAD3 in drought and salinity stress tolerance in soybean is lacking. We used bean pod mottle virus (BPMV)-based vector for achieving rapid and efficient overexpression as well as silencing of Omega-3 Fatty Acid Desaturase gene from Glycine max (GmFAD3) to assess the functional role of GmFAD3 in abiotic stress responses in soybean. Higher levels of recombinant BPMV-GmFAD3A transcripts were detected in overexpressing soybean plants. Overexpression of GmFAD3A in soybean resulted in increased levels of jasmonic acid and higher expression of GmWRKY54 as compared to mock-inoculated, vector-infected and FAD3-silenced soybean plants under drought and salinity stress conditions. The GmFAD3A-overexpressing plants showed higher levels of chlorophyll content, efficient photosystem-II, relative water content, transpiration rate, stomatal conductance, proline content and also cooler canopy under drought and salinity stress conditions as compared to mock-inoculated, vector-infected and FAD3-silenced soybean plants. Results from the current study revealed that GmFAD3A-overexpressing soybean plants exhibited tolerance to drought and salinity stresses. However, soybean plants silenced for GmFAD3 were vulnerable to drought and salinity stresses.

Small RNA profiling for identification of microRNAs involved in regulation of seed development and lipid biosynthesis in yellowhorn

BackgroundYellowhorn (Xanthoceras sorbifolium), an endemic woody oil-bearing tree, has become economically important and is widely cultivated in northern China for bioactive oil production. However, the regulatory mechanisms of seed development and lipid biosynthesis affecting oil production in yellowhorn are still elusive. MicroRNAs (miRNAs) play crucial roles in diverse aspects of biological and metabolic processes in seeds, especially in seed development and lipid metabolism. It is still unknown how the miRNAs regulate the seed development and lipid biosynthesis in yellowhorn.ResultsHere, based on investigations of differences in the seed growth tendency and embryo oil content between high-oil-content and low-oil-content lines, we constructed small RNA libraries from yellowhorn embryos at four seed development stages of the two lines and then profiled small RNA expression using high-throughput sequencing. A total of 249 known miRNAs from 46 families and 88 novel miRNAs were identified. Furthermore, by pairwise comparisons among the four seed development stages in each line, we found that 64 miRNAs (53 known and 11 novel miRNAs) were differentially expressed in the two lines. Across the two lines, 15, 11, 10, and 7 differentially expressed miRNAs were detected at 40, 54, 68, and 81 days after anthesis, respectively. Bioinformatic analysis was used to predict a total of 2654 target genes for 141 differentially expressed miRNAs (120 known and 21 novel miRNAs). Most of these genes were involved in the fatty acid biosynthetic process, regulation of transcription, nucleus, and response to auxin. Using quantitative real-time PCR and an integrated analysis of miRNA and mRNA expression, miRNA-target regulatory modules that may be involved in yellowhorn seed size, weight, and lipid biosynthesis were identified, such as miR172b-ARF2 (auxin response factor 2), miR7760-p3_1-AGL61 (AGAMOUS-LIKE 61), miR319p_1-FAD2–2 (omega-6 fatty acid desaturase 2–2), miR5647-p3_1-DGAT1 (diacylglycerol acyltransferase 1), and miR7760-p5_1-MED15A (Mediator subunit 15a).ConclusionsThis study provides new insights into the important regulatory roles of miRNAs in the seed development and lipid biosynthesis in yellowhorn. Our results will be valuable for dissecting the post-transcriptional and transcriptional regulation of seed development and lipid biosynthesis, as well as improving yellowhorn in northern China.

Reference Genes and Expression Analysis of Seed Desaturases Genes in Soybean Mutant Accessions.

Soybean oil is the second most-produced vegetable oil worldwide. To enhance the nutritional quality and oxidative stability of soybean oil, many soybean breeding programs are trying to increase oleic acid content and reduce linoleic and linolenic acid contents. The fatty acid profile of soybean oil is controlled by many genes, including those which code for omega-3 and omega-6 desaturases. Mutations in GmFAD2-1 and GmFAD3 genes are widely studied and their combinations can produce soybean oil with high oleic and low linoleic and linolenic content. However, few studies evaluate the effect of these mutations on gene expression. Therefore, the present study sought to identify reference genes, evaluate the expression of GmFAD2-1 and GmFAD3 seed desaturase genes in thirteen wild-type and mutated soybean accessions, and associate the expression patterns with fatty acid composition and with the GmFAD2-1 and GmFAD3 genotypes. GmCONS7 and GmUKN2 were identified as the best reference genes for combined use to normalize data. The GmFAD2-1A mutation of PI603452 accession was associated with a decrease in gene expression of GmFAD2-1A; however, downregulation may not be due to the truncated enzyme structure alone. These results suggested that there are factors other than GmFAD2-1A and GmFAD2-1B that have a considerable effect on oleic content, at least in soybeans with mutations in these two genes.

Dynamics of sugars, endogenous hormones, and oil content during the development of Camellia oleifera fruit

Camellia oleifera Abel (‘Huashuo’) is an important woody oil species with high economic value. Low yield is the main factor restricting the rapid development of the tea-oil industry. The dynamics of sugars, oil, endogenous hormones, and the genes involved in these pathways, as well as the correlation with oil synthesis-related factors in Huashuo, were investigated. The results showed that oil content was positively correlated with sucrose and starch content but was negatively correlated with reducing sugar content. Pyruvate kinase (PK) and fructose-1,6-bisphosphate aldolase (FBA) of the glycolysis pathway could be beneficial for oil biosynthesis. Higher levels of acetyl CoA carboxylase (ACC), stearoyl-ACP desaturase (SAD), and WRINKLED1 (WRI1), as well as lower levels of omega-6 fatty acid desaturase 2 (FAD2) and lipoxygenase 5 (LOX5) could be responsible for the oil production. The higher expression of gibberellin 20 oxidase (GA20ox), lower expression of gibberellic acid insensitive (GAI), and the increase in 9-cis-epoxycarotenoid dioxygenase (NCED) at the mRNA level could contribute to oil accumulation. The results of this study could provide a reference for increasing the yield of plants related to Huashuo.

Shotgun proteomics of peach fruit reveals major metabolic pathways associated to ripening

BackgroundFruit ripening in Prunus persica melting varieties involves several physiological changes that have a direct impact on the fruit organoleptic quality and storage potential. By studying the proteomic differences between the mesocarp of mature and ripe fruit, it would be possible to highlight critical molecular processes involved in the fruit ripening.ResultsTo accomplish this goal, the proteome from mature and ripe fruit was assessed from the variety O’Henry through shotgun proteomics using 1D-gel (PAGE-SDS) as fractionation method followed by LC/MS-MS analysis. Data from the 131,435 spectra could be matched to 2740 proteins, using the peach genome reference v1. After data pre-treatment, 1663 proteins could be used for comparison with datasets assessed using transcriptomic approaches and for quantitative protein accumulation analysis. Close to 26% of the genes that code for the proteins assessed displayed higher expression at ripe fruit compared to other fruit developmental stages, based on published transcriptomic data. Differential accumulation analysis between mature and ripe fruit revealed that 15% of the proteins identified were modulated by the ripening process, with glycogen and isocitrate metabolism, and protein localization overrepresented in mature fruit, as well as cell wall modification in ripe fruit. Potential biomarkers for the ripening process, due to their differential accumulation and gene expression pattern, included a pectin methylesterase inhibitor, a gibbellerin 2-beta-dioxygenase, an omega-6 fatty acid desaturase, a homeobox-leucine zipper protein and an ACC oxidase. Transcription factors enriched in NAC and Myb protein domains would target preferentially the genes encoding proteins more abundant in mature and ripe fruit, respectively.ConclusionsShotgun proteomics is an unbiased approach to get deeper into the proteome allowing to detect differences in protein abundance between samples. This technique provided a resolution so that individual gene products could be identified. Many proteins likely involved in cell wall and sugar metabolism, aroma and color, change their abundance during the transition from mature to ripe fruit.

In Silico Analysis of Partial Fatty Acid Desaturase 2 cDNA From Reutealis trisperma (Blanco) Airy Shaw.

Reutealis trisperma oil is a new source for biodiesel production. The predominant fatty acids in this plant are stearic acid (9%), palmitic acid (10%), oleic acid (12%), linoleic acid (19%), and α-eleostearic acid (51%). The presence of polyunsaturated fatty acids (PUFAs), linoleic acid, and α-eleostearic acid decreases the oxidation stability of R. trisperma biodiesel. Although several studies have suggested that the fatty acid desaturase 2 (FAD2) enzyme is involved in the regulation of fatty acid desaturation, little is known about the genetic information of FAD2 in R. trisperma. The objectives of this study were to isolate, characterize, and determine the relationship between the R. trisperma FAD2 fragment and other Euphorbiaceae plants. cDNA fragments were isolated using reverse transcription polymerase chain reaction (PCR). The DNA sequence obtained by sequencing was used for further analysis. In silico analysis identified the fragment identity, subcellular localization, and phylogenetic construction of the R. trisperma FAD2 cDNA fragment and Euphorbiaceae. The results showed that a 923-bp partial sequence of R. trisperma FAD2 was successfully isolated. Based on in silico analysis, FAD2 was predicted to encode 260 amino acids, had a domain similarity with Omega-6 fatty acid desaturase, and was located in the endoplasmic reticulum membrane. The R. trisperma FAD2 fragment was more closely related to Vernicia fordii (HM755946.1).

Effects of Salicylic Acid on Fatty Acid Gene Expression in Carthamus tinctorious L. cv. Dinçer under Pendimethalin Stress

Pendimethalin is a member of the dinitroaniline class herbicide. It used to control most annual grasses and many annual broad-leaved weeds. Salicylic acid acts as an endogenous signal molecule in charge of inducing environmental stress tolerance in plants. Omega-3 fatty acid desaturase is a key enzyme for α-linolenic acid biosynthesis. Here, we searched to understand the beneficial impacts of salicylic acid on fatty acid desaturase gene (FAD3 and FAD7) expression during pendimethalin stress in safflower (Carthamus tinctorious cv. “Dinçer). In this study, 0.004 and 0.01 M pendimethalin was applied to safflower plants grown under controlled climatic conditions in 36 pots. 0.05 mM salicylic acid was applied to the samples where pendimethalin was applied. After application, gene expression analyze were performed using quantitative polymerase chain reaction. The floor change between the test and control groups was calculated with the formula 2 ^ (delta delta CT). The change between gene expression levels was evaluated by t test (p <0.05). FAD3 and FAD7 expression levels decreased at low pendimethalin concentration. A down-regulation in FAD3 expression was observed in high pendimethalin stress, while an up-regulation in FAD7 expression was detected. Salicylic acid had a ameliorative effect on the negative effect of pendimethalin stress on FAD3 and FAD7 gene expression.

Transcriptome analysis of unsaturated fatty acids biosynthesis shows essential genes in sprouting of Acer truncatum Bunge seeds

The seeds of Acer truncatum Bunge (ATB) are rich in unsaturated fatty acids (UFA), especially nervonic acid, an important nutrient for the growth and maintenance of the brain. Therefore, the oil of ATB seeds has been approved as a New Resource Food in China. In the present study, changes of transcriptomic profiling and contents of UFA during the sprouting of ATB seeds were analyzed. The results of RNA-seq-based transcriptome analysis showed that the transcription of genes in various metabolic pathways, including the biosynthesis pathway of UFA, were significantly altered during seed sprouting. The gas chromatography-mass spectrometry assay consistently showed that the content of palmitoleic acid, palmitic acid, stearic acid, gondoic acid, arachidic acid, behenic acid, nervonic acid and lignoceric acid increased 77.9, 20.9, 39.6, 26.3, 95.6, 80.0, 41.7 and 141%, respectively, while the content of oleic acid decreased 49.2%, after sprouting. The whole coding sequence regions of genes involved in the UFA biosynthesis were cloned and sequenced, and the alignment analysis predicted that omega-6 fatty acid desaturase and short-chain type dehydrogenase/reductase might have important roles in the biosynthesis of UFA during the sprouting. The results suggested that nervonic acid could be enriched in ATB seeds via sprouting-stimulation, which had not been previously observed.

Fat-1 transgenic zebrafish are protected from abnormal lipid deposition induced by high-vegetable oil feeding.

High dietary concentration of vegetable oil, particularly those rich in n-6 polyunsaturated fatty acids (PUFAs), can induce negative physiological effects including excessive lipid deposition in teleost fish. Omega-3 desaturase (Fat-1) of Caenorhabditis elegans is able to convert n-6 PUFAs to n-3 PUFAs and thus induces a low n-6/n-3 PUFAs ratio alleviating lipid deposition. In this study, we investigated the effects of dietary n-6 PUFAs on lipid metabolism of fat-1 transgenic zebrafish (Tg:fat-1), to explore the role of fat-1 in fish lipid metabolism. We first generated Tg:fat-1 zebrafish and assayed the effects of a low-fat diet (LFD) and a high-fat diet (HFD) prepared from soybean oil. Wild type zebrafish (WT) fed with HFD (HFD-WT) exhibited increased obesity and lipid deposition, especially in the abdominal cavity and liver. These defects were absent from HFD-Tg:fat-1. For each diet group, Tg:fat-1 exhibited significantly decreased levels of almost all hepatic lipid classes compared with WT. Expression levels of lipid synthesis-related genes and lipid deposition-related genes were markedly lower in the liver of HFD-Tg:fat-1 compared with HFD-WT. In contrast, the steatolysis-related genes significantly upregulated in HFD-Tg:fat-1. Then expression profiles of mitochondrial energy metabolism-related genes and ATP contents in the livers from LFD-WT, LFD-Tg:fat-1, HFD-WT, and HFD-Tg:fat-1 were determined. Our findings suggest that fat-1 protects fish from abnormal lipid deposition induced by high-vegetable oil feeding, through endogenously converting n-6 PUFAs to n-3 PUFAs. KEY POINTS: • fat-1 transgenic zebrafish (Tg:fat-1) can endogenously convert n-6 PUFAs to n-3 PUFAs. • Tg:fat-1 avoid serious abnormal lipid deposition induced by high-vegetable oil feeding. • fat-1 transgenosis effectively improved lipid metabolism and mitochondrial energy metabolism in zebrafish.

Transcriptome Analysis of High-NUE (T29) and Low-NUE (T13) Genotypes Identified Different Responsive Patterns Involved in Nitrogen Stress in Ramie (Boehmeria nivea (L.) Gaudich).

Nitrogen-use efficiency (NUE) has significant impacts on plant growth and development. NUE in plants differs substantially in physiological resilience to nitrogen stress; however, the molecular mechanisms underlying enhanced resilience of high-NUE plants to nitrogen deficiency remains unclear. We compared transcriptome-wide gene expression between high-NUE and low-NUE ramie (Boehmeria nivea (L.) Gaudich) genotypes under nitrogen (N)-deficient and normal conditions to identify the transcriptomic expression patterns that contribute to ramie resilience to nitrogen deficiency. Two ramie genotypes with contrasting NUE were used in the study, including T29 (NUE = 46.01%) and T13 (NUE = 15.81%). Our results showed that high-NUE genotypes had higher gene expression under the control condition across 94 genes, including frontloaded genes such as GDSL esterase and lipase, gibberellin, UDP-glycosyltransferase, and omega-6 fatty acid desaturase. Seventeen stress-tolerance genes showed lower expression levels and varied little in response to N-deficiency stress in high-NUE genotypes. In contrast, 170 genes were upregulated under N deficiency in high-NUE genotypes but downregulated in low-NUE genotypes compared with the controls. Furthermore, we identified the potential key genes that enable ramie to maintain physiological resilience under N-deficiency stress, and categorized these genes into three groups based on the transcriptome and their expression patterns. The transcriptomic and clustering analysis of these nitrogen-utilization-related genes could provide insight to better understand the mechanism of linking among the three gene classes that enhance resilience in high-NUE ramie genotypes.

Exogenous Abscisic Acid Supplementation at Early Stationary Growth Phase Triggers Changes in the Regulation of Fatty Acid Biosynthesis in Chlorella vulgaris UMT-M1.

Abscisic acid (ABA) has been known to exist in a microalgal system and serves as one of the chemical stimuli in various biological pathways. Nonetheless, the involvement of ABA in fatty acid biosynthesis, particularly at the transcription level in microalgae is poorly understood. The objective of this study was to determine the effects of exogenous ABA on growth, total oil content, fatty acid composition, and the expression level of beta ketoacyl-ACP synthase I (KAS I) and omega-3 fatty acid desaturase (ω-3 FAD) genes in Chlorella vulgaris UMT-M1. ABA was applied to early stationary C. vulgaris cultures at concentrations of 0, 10, 20, and 80μM for 48h. The results showed that ABA significantly increased biomass production and total oil content. The increment of palmitic (C16:0) and stearic (C18:0) acids was coupled by decrement in linoleic (C18:2) and α-linolenic (C18:3n3) acids. Both KAS I and ω-3 FAD gene expression were downregulated, which was negatively correlated to saturated fatty acid (SFAs), but positively correlated to polyunsaturated fatty acid (PUFA) accumulations. Further analysis of both KAS I and ω-3 FAD promoters revealed the presence of multiple ABA-responsive elements (ABREs) in addition to other phytohormone-responsive elements. However, the role of these phytohormone-responsive elements in regulating KAS I and ω-3 FAD gene expression still remains elusive. This revelation might suggest that phytohormone-responsive gene regulation in C. vulgaris and microalgae as a whole might diverge from higher plants which deserve further scientific research to elucidate its functional roles.

Exogenous methylglyoxal enhances the reactive aldehyde detoxification capability and frost-hardiness of wheat

Cold-acclimation is essential for the development of adequate frost-hardiness in cereals and therefore sudden freezes can cause considerable damage to the canopy. However, timely adding of an appropriate signal in the absence of cold acclimation may also harden wheat for the upcoming freeze. The feasibility of the promising signal molecule methylglyoxal was tested here for such applications and the signal mechanism was studied in bread wheat (Triticum aestivum L.) and durum wheat (Triticum turgidum L. ssp. durum). Spraying with 10 mM methylglyoxal did not decrease the fresh weight and photosynthetic parameters in most wheat varieties at growth temperature (21 °C). Photosynthetic parameters even improved and chlorophyll content increased in some cases. Increased transcript level of glutathione-S-transferases and omega-3 fatty acid desaturases was detected by qPCR 6 h after the last methylglyoxal spray. Aldo-keto reductase and glyoxalase enzyme activities, as well as sorbitol content of wheat plants increased 24 h after the last 10 mM methylglyoxal spray in most of the cultivars. These mechanisms may explain the increased freezing survival of methylglyoxal pretreated wheat plants from less than 10% to over 30%. Our results demonstrate that exogenous methylglyoxal treatment can be safely added to wheat plants as preparatory treatment without detrimental effects but inducing some of the stress-protective mechanisms, which contribute to frost-hardiness.

Influence of nitrogen availability on biomass, lipid production, fatty acid profile, and the expression of fatty acid desaturase genes in Messastrum gracile SE-MC4.

In this study, the effects of limited and excess nitrate on biomass, lipid production, and fatty acid profile in Messastrum gracile SE-MC4 were determined. The expression of fatty acid desaturase genes, namely stearoyl-ACP desaturase (SAD), omega-6 fatty acid desaturase (ω-6 FAD), omega-3 fatty acid desaturase isoform 1 (ω-3 FADi1), and omega-3 fatty acid desaturase isoform 2 (ω-3 FADi2) was also assessed. It was found that nitrate limitation generally increased the total oil, α-linolenic acid (C18:3n3) and total polyunsaturated fatty acid (PUFA) contents in M. gracile. The reduction of nitrate concentration from 1.76 to 0.11mM increased the total oil content significantly from 32.5 to 41.85% (dry weight). Palmitic (C16:0) and oleic (C18:1) acids as the predominant fatty acids in this microalgae constituted between 82 and 87% of the total oil content and were relatively consistent throughout all nitrate concentrations tested. The expression of SAD, ω-6 FAD, and ω-3 FADi2 genes increased under nitrate limitation, especially at 0.11mM nitrate. The ω-3 FADi1 demonstrated a binary up-regulation pattern of expression under both nitrate-deficient (0.11mM) and -excess (3.55mM) conditions. Thus, findings from this study suggested that limited or excess nitrate could be used as part of a cultivation strategy to increase oil and PUFA content following media optimisation and more efficient culture methodology. Data obtained from the expression of desaturase genes would provide valuable insights into their roles under excess and limited nitrate conditions in M. gracile, potentially paving the way for future genetic modifications.

Effects of Excess and Limited Phosphate on Biomass, Lipid and Fatty Acid Contents and the Expression of Four Fatty Acid Desaturase Genes in the Tropical Selenastraceaen Messastrum gracile SE-MC4.

In this study, the effects of limited and excess phosphate on biomass content, oil content, fatty acid profile and the expression of three fatty acid desaturases in Messastrum gracile SE-MC4 were determined. It was found that total biomass (0.67-0.83gL-1), oil content (30.99-38.08%) and the duration for cells to reach stationary phase (25-27days) were not considerably affected by phosphate limitation. However, excess phosphate slightly reduced total biomass and oil content to 0.50gL-1 and 25.36% respectively. The dominant fatty acids in M. gracile, pamitic acid (C16:0) and oleic acid (C18:1) which constitute more than 81% of the total fatty acids remained relatively high and constant across all phosphate concentrations. Reduction of phosphate concentration to 25% and below significantly increased total MUFA, whereas increasing phosphate concentration to ≥ 50% and ≥ 100% significantly increased total SFA and PUFA content respectively. The expression of omega-3 fatty acid desaturase (ω-3 FADi1, ω-3 FADi2) and omega-6 fatty acid desaturase (ω-6 FAD) was increased under phosphate limitation, especially at ≤ 12.5% phosphate, whereas levels of streoyl-ACP desaturase (SAD) transcripts were relatively unchanged across all phosphate concentrations. The first isoform of ω-3 FAD (ω-3 FADi) displayed a binary upregulation under limited (≤ 12.5%) and excess (200%) phosphate. The expression of ω-6 FAD, ω-3 FAD and SAD were inconsistent with the accumulation of oleic acid (C18:1), linoleic acid (C18:2) and alpha-linolenic acid (C18:3), suggesting that these genes may be regulated indirectly by phosphate availability via post-transcriptional or post-translational mechanisms.

ITRAQ-based proteomics reveals key role of γ-aminobutyric acid (GABA) in regulating drought tolerance in perennial creeping bentgrass (Agrostis stolonifera)

γ-Aminobutyric acid (GABA), a non-proteinaceous amino acid, modulates plant growth and stress tolerance. However, the potential role of GABA in regulating key metabolic pathways and stress-defensive proteins against drought in plants has never been explored. Creeping bentgrass (Agrostis stolonifera) plants were pretreated with or without GABA and then subjected to water stress for 8 days in controlled growth chambers (23/19 °C, day/night). Physiological analysis showed that elevated endogenous GABA level via exogenous GABA application significantly mitigated water stress damage to creeping bentgrass, as manifested by increased leaf relative water content, water use efficiency, osmotic adjustment (OA), photochemical efficiency (Fv/Fm), net photosynthetic rate, and reduced oxidative damage. iTRAQ-based proteomics found that enhanced chaperones accumulation, carbohydrates, amino acids, and energy metabolism played important roles in protein protection, OA, energy maintenance, and metabolic balance, which is important adaptive response to drought stress in creeping bentgrass. The GABA further promoted energy production and conversion, antioxidant defense, and DHN3 accumulation that were essential for energy requirement, ROS-scavenging, and the prevention of cell dehydration in leaf during drought stress. In addition, GABA-treated plants maintained significantly higher abundance of dicarboxylate transporter 2.1, ATP-dependent zinc metalloprotease, receptor-like protein kinase HERK1, o-acyltransferase WSD1, omega-6 fatty acid desaturase, and two-component response regulator ORR21 than untreated plants under drought stress. The result provides new evidences that GABA-induced drought tolerance is possibly involved in the improvement of nitrogen recycling, protection of photosystem II, mitigation of drought-depressed cell elongation, wax biosynthesis, fatty acid desaturase, and delaying leaf senescence in creeping bentgrass.