scholarly journals Cloning and Functional Analysis of the FAD2 Gene Family from Desert Shrub Artemisia sphaerocephala

2019 ◽  
Author(s):  
Xiumei Miao ◽  
lijing zhang ◽  
Xiaowei Hu ◽  
Shuzhen Nan ◽  
Xiaolong Chen ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Linoleic Acid ◽  
Gene Family ◽  
Family Members ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
Acid Biosynthesis ◽  
Desert Shrub ◽  
Fad2 Gene ◽  
Artemisia Sphaerocephala

Abstract Background: Linoleic acid is an important polyunsaturated fatty acid, required for all eukaryotes. Microsomal delta-12 (Δ12) oleate desaturase (FAD2) is a key enzyme for linoleic acid biosynthesis. Desert shrub Artemisia sphaerocephala is rich in linoleic acid, it has a large FAD2 gene family with twenty-six members. The objective of this work is to study the difference and potentially functionality of AsFAD2 family members. Results: Full-length cDNAs of twenty-one AsFAD2 genes were obtained from A. sphaerocephala. The putative polypeptides encoded by AsFAD2 family genes showed a high level of sequence similarity and were relatively conserved during evolution. The motif composition was relatively conservative. Quantitative real-time PCR analysis revealed that the AsFAD2-1 gene was strongly expressed in developing seeds, which may be closely associated with the high accumulating ability of linoleic acid in A. sphaerocephala seeds. Although different AsFAD2 family members showed diverse response to salt stress, the overall mRNA levels of the AsFAD2 family genes was stable, suggesting that the AsFAD2 family may play an important role in the adaptation of A. sphaerocephala to the harsh desert environment. Transient expression of AsFAD2 genes in the Nicotiana benthamiana leaves revealed that the encoded proteins were all located in the endoplasmic reticulum. Heterologous expression in Saccharomyces cerevisiae suggested that only three AsFAD2 enzymes, AsFAD2-1, -10, and -23, were Δ12 oleate desaturases, which could convert oleic acid to linoleic acid, whereas AsFAD2-1 and AsFAD2-10 could also produce palmitolinoleic acid. Conclusions: This research reported the cloning, expression studies, subcellular localization and functional identification of the large AsFAD2 gene family. These results should be helpful in understanding of fatty acid biosynthesis in A. sphaerocephala, and can be applied in the study of plant fatty acids traits.

scholarly journals Cloning and Functional Analysis of the FAD2 Gene Family from Desert Shrub Artemisia sphaerocephala

2019 ◽  
Author(s):  
Xiumei Miao ◽  
lijing zhang ◽  
Xiaowei Hu ◽  
Shuzhen Nan ◽  
Xiaolong Chen ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Linoleic Acid ◽  
Gene Family ◽  
Family Members ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
Acid Biosynthesis ◽  
Desert Shrub ◽  
Fad2 Gene ◽  
Artemisia Sphaerocephala

Abstract Background: Linoleic acid is an important polyunsaturated fatty acid, required for all eukaryotes. Microsomal delta-12 (Δ12) oleate desaturase (FAD2) is a key enzyme for linoleic acid biosynthesis. Desert shrub Artemisia sphaerocephala is rich in linoleic acid, it has a large FAD2 gene family with twenty-six members. The aim of this work is to unveil the difference and potentially functionality of AsFAD2 family members. Results: Full-length cDNAs of twenty-one AsFAD2 genes were obtained from A. sphaerocephala. The putative polypeptides encoded by AsFAD2 family genes showed a high level of sequence similarity and were relatively conserved during evolution. The motif composition was also relatively conservative. Quantitative real-time PCR analysis revealed that the AsFAD2-1 gene was strongly expressed in developing seeds, which may be closely associated with the high accumulating ability of linoleic acid in A. sphaerocephala seeds. Although different AsFAD2 family members showed diverse response to salt stress, the overall mRNA levels of the AsFAD2 family genes was stable. Transient expression of AsFAD2 genes in the Nicotiana benthamiana leaves revealed that the encoded proteins were all located in the endoplasmic reticulum. Heterologous expression in Saccharomyces cerevisiae suggested that only three AsFAD2 enzymes, AsFAD2-1, -10, and -23, were Δ12 oleate desaturases, which could convert oleic acid to linoleic acid, whereas AsFAD2-1 and AsFAD2-10 could also produce palmitolinoleic acid. Conclusions: This research reported the cloning, expression studies, subcellular localization and functional identification of the large AsFAD2 gene family. These results should be helpful in understanding fatty acid biosynthesis in A. sphaerocephala, and has the potential to be applied in the study of plant fatty acids traits.

scholarly journals Cloning and Functional Analysis of the FAD2 Gene Family from Desert Shrub Artemisia sphaerocephala

2019 ◽  
Author(s):  
Xiumei Miao ◽  
lijing zhang ◽  
Xiaowei Hu ◽  
Shuzhen Nan ◽  
Xiaolong Chen ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Linoleic Acid ◽  
Gene Family ◽  
Family Members ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
Acid Biosynthesis ◽  
Desert Shrub ◽  
Fad2 Gene ◽  
Artemisia Sphaerocephala

Abstract Background: Linoleic acid is an important polyunsaturated fatty acid, required for all eukaryotes. Microsomal delta-12 (Δ12) oleate desaturase (FAD2) is a key enzyme for linoleic acid biosynthesis. Desert shrub Artemisia sphaerocephala is rich in linoleic acid, it has a large FAD2 gene family with twenty-six members. The aim of this work is to unveil the difference and potentially functionality of AsFAD2 family members. Results: Full-length cDNAs of twenty-one AsFAD2 genes were obtained from A. sphaerocephala. The putative polypeptides encoded by AsFAD2 family genes showed a high level of sequence similarity and were relatively conserved during evolution. The motif composition was also relatively conservative. Quantitative real-time PCR analysis revealed that the AsFAD2-1 gene was strongly expressed in developing seeds, which may be closely associated with the high accumulating ability of linoleic acid in A. sphaerocephala seeds. Although different AsFAD2 family members showed diverse response to salt stress, the overall mRNA levels of the AsFAD2 family genes was stable. Transient expression of AsFAD2 genes in the Nicotiana benthamiana leaves revealed that the encoded proteins were all located in the endoplasmic reticulum. Heterologous expression in Saccharomyces cerevisiae suggested that only three AsFAD2 enzymes, AsFAD2-1, -10, and -23, were Δ12 oleate desaturases, which could convert oleic acid to linoleic acid, whereas AsFAD2-1 and AsFAD2-10 could also produce palmitolinoleic acid. Conclusions: This research reported the cloning, expression studies, subcellular localization and functional identification of the large AsFAD2 gene family. These results should be helpful in understanding fatty acid biosynthesis in A. sphaerocephala, and has the potential to be applied in the study of plant fatty acids traits.

scholarly journals Cloning and functional analysis of the FAD2 gene family from desert shrub Artemisia sphaerocephala

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Xiumei Miao ◽  
Lijing Zhang ◽  
Xiaowei Hu ◽  
Shuzhen Nan ◽  
Xiaolong Chen ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Linoleic Acid ◽  
Gene Family ◽  
Fatty Acid Biosynthesis ◽  
Family Members ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
Acid Biosynthesis ◽  
Desert Shrub ◽  
Artemisia Sphaerocephala

Abstract Background Linoleic acid is an important polyunsaturated fatty acid, required for all eukaryotes. Microsomal delta-12 (Δ12) oleate desaturase (FAD2) is a key enzyme for linoleic acid biosynthesis. Desert shrub Artemisia sphaerocephala is rich in linoleic acid, it has a large FAD2 gene family with twenty-six members. The aim of this work is to unveil the difference and potentially functionality of AsFAD2 family members. Results Full-length cDNAs of twenty-one AsFAD2 genes were obtained from A. sphaerocephala. The putative polypeptides encoded by AsFAD2 family genes showed a high level of sequence similarity and were relatively conserved during evolution. The motif composition was also relatively conservative. Quantitative real-time PCR analysis revealed that the AsFAD2–1 gene was strongly expressed in developing seeds, which may be closely associated with the high accumulating ability of linoleic acid in A. sphaerocephala seeds. Although different AsFAD2 family members showed diverse response to salt stress, the overall mRNA levels of the AsFAD2 family genes was stable. Transient expression of AsFAD2 genes in the Nicotiana benthamiana leaves revealed that the encoded proteins were all located in the endoplasmic reticulum. Heterologous expression in Saccharomyces cerevisiae suggested that only three AsFAD2 enzymes, AsFAD2–1, − 10, and − 23, were Δ12 oleate desaturases, which could convert oleic acid to linoleic acid, whereas AsFAD2–1 and AsFAD2–10 could also produce palmitolinoleic acid. Conclusions This research reported the cloning, expression studies, subcellular localization and functional identification of the large AsFAD2 gene family. These results should be helpful in understanding fatty acid biosynthesis in A. sphaerocephala, and has the potential to be applied in the study of plant fatty acids traits.

Insights into the novel members of the FAD2 gene family involved in high-oleate fluxes in peanut

Genome ◽  
2015 ◽  
Vol 58 (8) ◽  
pp. 375-383 ◽  
Author(s):  
Yun Wang ◽  
Xingguo Zhang ◽  
Yongli Zhao ◽  
C.S. Prakash ◽  
Guohao He ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Linoleic Acid ◽  
Gene Family ◽  
Gene Networks ◽  
Genetic Manipulation ◽  
Oil Quality ◽  
Fatty Acid Desaturase ◽  
Sequence Comparisons ◽  
Developing Seeds ◽  
Fad2 Gene

The FAD2 gene family is functionally responsible for the conversion of oleic acid to linoleic acid in oilseed plants. Multiple members of the FAD gene are known to occur in several oilseed species. In this study, six novel full-length cDNA sequences (named as AhFAD2-1, -2, -3, -4, -5, and -6) were identified in peanut (Arachis hypogaea L.), an analysis of which revealed open reading frames of 379, 383, 394, or 442 amino acids. Sequence comparisons showed that AhFAD2-1 and AhFAD2-2 shared 76% identity, while AhFAD2-2, -3, and -4 displayed highly significant homology. There was only 27% identity overlap between the microsomal ω-6 fatty acid desaturase and the chloroplast ω-6 fatty acid desaturase encoded by AhFAD2-1, -2, -3, -4, and AhFAD2-5, -6, respectively. The phylogeny tree of FAD2 transcripts showed five major groups, and AhFAD2-1 was clearly separated from other groups. Analysis of AhFAD2-1 and AhFAD2-2 transcript distribution in different peanut tissues showed that the AhFAD2-1 gene showed upward of a 70-fold increase in expression of fatty acid than the AhFAD2-2 gene in peanut developing seeds, while the AhFAD2-2 gene expressed most abundantly in peanut flowers. Because the AhFAD2-1 gene played a major role in the conversion of oleic to linoleic acid during seed development, the identification of this novel member in this study would facilitate the further genetic manipulation of peanut oil quality. The implications of overall results also suggest that there may be more candidate genes controlling levels of oleate acid in developing seeds. Results also may be due to the presence of complex gene networks controlling the fluxes between the endoplasmic reticulum and the chloroplast within the peanut cells.

scholarly journals Specialized Functions of Olive FAD2 Gene Family Members Related to Fruit Development and the Abiotic Stress Response

2019 ◽  
Vol 61 (2) ◽  
pp. 427-441 ◽  
Author(s):  
M Luisa Hernández ◽  
M Dolores Sicardo ◽  
Patricia M Arjona ◽  
José M Martínez-Rivas
Keyword(s):  
Gene Expression ◽  
Linoleic Acid ◽  
Gene Family ◽  
Olive Oil ◽  
Membrane Lipids ◽  
Lipid Analysis ◽  
Family Members ◽  
Functional Expression ◽  
Abiotic Stress Response ◽  
Fad2 Gene

Abstract Three different cDNA sequences, designated OepFAD2-3, OepFAD2-4 and OepFAD2-5, encoding three microsomal oleate desaturases (FAD2) have been isolated from olive (Olea europaea cv. Picual). Sequence analysis and functional expression in yeast of the corresponding cDNAs confirm that they encode microsomal oleate desaturases. Gene expression and lipid analysis indicate that these three genes are not involved in the linoleic acid present in seed lipids, while OeFAD2-5, together with OeFAD2-2, contributes mostly to the linoleic acid present in the mesocarp and, therefore, in the olive oil. Our results have also shown that olive FAD2-3, FAD2-4 and FAD2-5 gene expression is not only spatially and temporally regulated in olive fruit, but also is cultivar-dependent, as well as regulated by water regime, temperature, light and wounding. All these data suggest specialized physiological roles for the olive FAD2 gene family members with respect to both aspects of the biosynthesis of the linoleic acid, either present in storage lipids that constitute the olive oil or being part of membrane lipids, which are involved in the response to abiotic stresses, and highlight the differences on FAD2 gene regulation between oilseeds and oil fruits.

Differential regulation of fatty acid elongation enzymes in brown adipocytes implies a unique role for Elovl3 during increased fatty acid oxidation

2005 ◽  
Vol 289 (4) ◽  
pp. E517-E526 ◽  
Author(s):  
Andreas Jakobsson ◽  
Johanna A. Jörgensen ◽  
Anders Jacobsson
Keyword(s):  
Fatty Acid ◽  
Gene Family ◽  
Regulatory Element ◽  
Saturated Fatty Acids ◽  
Acid Oxidation ◽  
Peroxisome Proliferator ◽  
Mrna Levels ◽  
Cold Stimulation ◽  
Brown Adipocytes ◽  
Brown Adipose

The expression of the Elovl3 gene, which belongs to the Elovl gene family coding for microsomal enzymes involved in very long-chain fatty acid (VLCFA) elongation, is dramatically increased in mouse brown adipose tissue upon cold stimulation. In the present study, we show that the cold-induced Elovl3 expression is under the control of peroxisome proliferator-activated receptor-α (PPARα) and that this regulation is part of a fundamental divergence in the regulation of expression for the different members of the Elovl gene family. In cultured brown adipocytes, a mixture of norepinephrine, dexamethasone, and the PPARα ligand Wy-14643, which rendered the adipocytes a high oxidative state, was required for substantial induction of Elovl3 expression, whereas the same treatment suppressed Elovl1 mRNA levels. The nuclear liver X receptor (LXR) has been implicated in the control of fatty acid synthesis and subsequent lipogenic processes in several tissues. This regulation is also exerted in part by sterol regulatory element-binding protein (SREBP-1), which is a target gene of LXR. We found that stimulation of Elovl3 expression was independent of LXR and SREBP-1 activation. In addition, exposure to the LXR agonist TO-901317 increased nuclear abundance of LXR and mature SREBP-1 as well as expression of the elongases Lce and Elovl1 in a lipogenic fashion but repressed Elovl3 expression. A functional consequence of this was seen on the level of esterified saturated fatty acids, such as C22:0, which was coupled to Elovl3 expression. These data demonstrate differential transcriptional regulation and concomitantly different functional roles for fatty acid elongases in lipid metabolism of brown adipocytes, which reflects the metabolic status of the cells.

scholarly journals Maternal conjugated linoleic acid modulates TAG metabolism in adult rat offspring

2017 ◽  
Vol 118 (11) ◽  
pp. 906-913 ◽  
Author(s):  
Jimena Lavandera ◽  
Carolina D. Gerstner ◽  
Juliana Saín ◽  
Ana C. Fariña ◽  
Marcela A. González ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Fatty Acid ◽  
Linoleic Acid ◽  
Conjugated Linoleic Acid ◽  
Fatty Acid Synthase ◽  
Regulatory Element ◽  
Male Rat ◽  
Mrna Levels ◽  
Fat Depots ◽  
Rat Offspring

AbstractConjugated linoleic acid (CLA) might regulate the lipid depots in liver and adipose tissue. As there is an association between maternal nutrition, fat depots and risk of offspring chronic disease, the aim was to investigate the effect of maternal CLA consumption on TAG regulation and some inflammatory parameters in adult male rat offspring receiving or not receiving CLA. Female Wistar rats were fed control (C) or CLA-supplemented (1 %, w/w) diets during 4 weeks before and throughout pregnancy and lactation. After weaning, male offspring of CLA rats were fed C or CLA diets (CLA/C and CLA/CLA groups, respectively), whereas C male rat offspring were fed a C diet (C/C group) for 9 weeks. Serum TAG levels were increased in the CLA/CLA and CLA/C groups, associated with a reduction of lipoprotein lipase activity and weights of adipose tissue. The liver TAG levels were decreased in the CLA/CLA group, related to a significant reduction of fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC) and glucose-6-phosphate dehydrogenase enzyme activities, as well as to the mRNA levels of FAS, ACC, stearoyl-CoA desaturase-1 and sterol regulatory element-binding protein-1c. Even though normal TAG levels were found in the liver of CLA/C rats, a reduction of lipogenesis was also observed. Thus, these results demonstrated a programming effect of CLA on the lipid metabolic pathways leading to a preventive effect on the TAG accretion in adipose tissue and the liver of male rat offspring. This knowledge could be important to develop some dietary strategies leading to a reduced incidence of obesity and fatty acid liver disease in humans.

scholarly journals Abscisic Acid Improves Linoleic Acid Accumulation Possibly by Promoting Expression of EgFAD2 and Other Fatty Acid Biosynthesis Genes in Oil Palm Mesocarp

2021 ◽  
Vol 12 ◽  
Author(s):  
Peng Shi ◽  
Wei Hua ◽  
Yin Min Htwe ◽  
Dapeng Zhang ◽  
Jun Li ◽  
...  
Keyword(s):  
Abscisic Acid ◽  
Fatty Acid ◽  
Linoleic Acid ◽  
Oil Palm ◽  
Fatty Acid Biosynthesis ◽  
Enrichment Analysis ◽  
Linoleic Acid Content ◽  
Pathway Enrichment Analysis ◽  
Acid Biosynthesis ◽  
Palm Fruit

Abscisic acid plays an important role in fruit development. However, the effect of ABA on fatty acid biosynthesis in oil palm is still unknown. In this study, ABA treatments (CK, A1–A4) were applied to oil palm fruit at 16 WAP (weeks after pollination), and fatty acids in the mesocarp at 24 WAP were analyzed by GC-MS. Results showed that linoleic acid content under treatment A2 (20 μM ABA) was significantly higher (slightly increased by 8.33%) than the control. Therefore, mesocarp samples of A2, and the control at 16, 20, and 24 WAP was sampled for RNA-Seq. KEGG pathway enrichment analysis showed that 43 genes were differentially expressed in the fatty acid biosynthesis pathway, of which expression of EgFAD2 (unigene 105050201) under 20 μM ABA treatment was 1.84-fold higher than in the control at 20 WAP. Further sequence analysis found that unigene 105050201 had more ABA-responsive elements (ABRE), complete conserved domains, and a C-terminal signaling motif among two FAD2 copies. Furthermore, WGCNA and correlation analysis showed co-expression of EgFAD2 (unigene 105050201) with transcription factors (TFs) (WRI1, AP2-EREBP, bZIP, bHLH, C2C2-Dof, MYB, NAC, and WRKY), ABA signaling genes (PYR, PP2C, SnRK, and ABI5), and other genes involved in fatty acid biosynthesis (FATA, FATB, LACS, SAD, Oleosins, and so on). These results indicated that ABA treatment promoted the expression of FAD2 and other genes involved in fatty acid biosynthesis, which possibly resulted in the accumulation of linoleic acid. This study will be helpful for understanding the possible mechanisms through which ABA affects fatty acid biosynthesis and their accumulation in the mesocarp of oil palm.

scholarly journals Identification and expression analysis of bovine ANGPTL gene family

2010 ◽  
Vol 56 (4) ◽  
pp. 445-453 ◽  
Author(s):  
Zhengbing Guan ◽  
Guolin Cai ◽  
Junyong Sun ◽  
Jian Lu
Keyword(s):  
Gene Family ◽  
Expression Analysis ◽  
Biological Activities ◽  
Expression Patterns ◽  
Family Members ◽  
Coiled Coil ◽  
Amino Acid Sequences ◽  
Pcr Analysis ◽  
Important Species ◽  
Real Time Quantitative Pcr

Abstract Encoded by seven genes, angiopoietin-like (ANGPTL) family members structurally similar to the angiogenic regulating factor angiopoietin are known to possess biological activities in angiogenesis and metabolism. Here we reports for the first time the identification and expression analysis of all the seven members of bovine ANGPTL gene family, which were designated bANGPTL1 to bANGPTL7 in order. The seven bANGPTL genes consist of 4-9 exons, span 3800-43000 bp and are located on different chromosomes. The deduced amino acid sequences of the members all possess an N-terminal coiled-coil domain and a C-terminal fibrinogen-like domain, both characteristics of angiopoietins. Phylogenetic analysis showed that the 32 identified ANGPTL homologs from 9 species could be classified into two major groups. Real-time quantitative PCR (Q-PCR) analysis revealed that the bANGPTL family members have different expression patterns. This study will be helpful for investigation on the biological role of the bANGPTL family in this economically important species. Furthermore, it provides an insight into the molecular evolution of the emerging ANGPTL family.

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