scholarly journals Isolation and functional analyses of PvFAD2 and PvFAD3 involved in the biosynthesis of polyunsaturated fatty acids from Sacha Inchi (Plukenetia volubilis)

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9169
Author(s):  
Tianquan Yang ◽  
Xiaojuan Wang ◽  
Tingnan Dong ◽  
Wei Xu ◽  
Aizhong Liu

The development of ω-3 fatty acid-rich vegetable oils is essential to enrich the production of functional foods. Sacha Inchi (Plukenetia volubilis L.) is a unique oilseed crop with much potential. Its seeds contain rich polyunsaturated fatty acids (PUFAs), especially linoleic acid (LA, C18:2) and α-linolenic acid (ALA, C18:3). Endoplasmic reticulum -located ω-6 and ω-3 fatty acid desaturases (FAD) are responsible for the biosynthesis of LA and ALA, respectively, in plant seeds. Here, we isolated two full-length FAD genes from Sacha Inchi, named PvFAD2 and PvFAD3, which encoded predicted amino acid residues of 384 and 379 in protein, respectively. Protein sequence and subcellular localization analysis revealed that they were located in the endoplasmic reticulum (ER). Heterologous expression in Saccharomyces cerevisiae confirmed that PvFAD2 and PvFAD3 could catalyze LA and ALA synthesis, respectively. The stability and catalytic efficiency of the PvFAD3 protein may be closely related to temperature. In transgenic tobacco, using seed-specific expression promoters, PvFAD2 and PvFAD3 significantly promotes the production of LA (from 68% to 70.5%) and ALA (from 0.7% to 3.1%) in seed oil. These results show that PvFAD2 and PvFAD3 do, indeed, function as crucial enzymes for PUFAs biosynthesis, and provide a key gene source for the sustainable production of lipids with tailored fatty acid compositions via genetic engineering in other oil crops.

1978 ◽  
Vol 174 (2) ◽  
pp. 585-593 ◽  
Author(s):  
Catherine T. Hammer ◽  
Eric D. Wills

The fatty acid compositions of the lipids and the lipid peroxide concentrations and rates of lipid peroxidation were determined in suspensions of liver endoplasmic reticulum isolated from rats fed on synthetic diets in which the fatty acid composition had been varied but the remaining constituents (protein, carbohydrate, vitamins and minerals) kept constant. Stock diet and synthetic diets containing no fat, 10% corn oil, herring oil, coconut oil or lard were used. The fatty acid composition of the liver endoplasmic reticulum lipid was markedly dependent on the fatty acid composition of the dietary lipid. Feeding a herring-oil diet caused incorporation of 8.7% eicosapentaenoic acid (C20:5) and 17% docosahexaenoic acid (C22:6), but only 5.1% linoleic acid (C18:2) and 6.4% arachidonic acid (C20:4), feeding a corn-oil diet caused incorporation of 25.1% C18:2, 17.8% C20:4 and 2.5% C22:6 fatty acids, and feeding a lard diet caused incorporation of 10.3% C18:2, 13.5% C20:4 and 4.3% C22:6 fatty acids into the liver endoplasmic-reticulum lipids. Phenobarbitone injection (100mg/kg) decreased the incorporation of C20:4 and C22:6 fatty acids into the liver endoplasmic reticulum of rats fed on a lard, corn-oil or herring-oil diet. Microsomal lipid peroxide concentrations and rates of peroxidation in the presence of ascorbate depended on the nature and quantity of the polyunsaturated fatty acids in the diet. The lipid peroxide content was 1.82±0.30nmol of malonaldehyde/mg of protein and the rate of peroxidation was 0.60±0.08nmol of malonaldehyde/min per mg of protein after feeding a fat-free diet, and the values were increased to 20.80nmol of malonaldehyde/mg of protein and 3.73nmol of malonaldehyde/min per mg of protein after feeding a 10% herring-oil diet in which polyunsaturated fatty acids formed 24% of the total fatty acids. Addition of α-tocopherol to the diets (120mg/kg of diet) caused a very large decrease in the lipid peroxide concentration and rate of lipid peroxidation in the endoplasmic reticulum, but addition of the synthetic anti-oxidant 2,6-di-t-butyl-4-methylphenol to the diet (100mg/kg of diet) was ineffective. Treatment of the animals with phenobarbitone (1mg/ml of drinking water) caused a sharp fall in the rate of lipid peroxidation. It is concluded that the polyunsaturated fatty acid composition of the diet regulates the fatty acid composition of the liver endoplasmic reticulum, and this in turn is an important factor controlling the rate and extent of lipid peroxidation in vitro and possibly in vivo.


2013 ◽  
Vol 141 (3) ◽  
pp. 1732-1739 ◽  
Author(s):  
Rosana Chirinos ◽  
Gledy Zuloeta ◽  
Romina Pedreschi ◽  
Eric Mignolet ◽  
Yvan Larondelle ◽  
...  

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Hung Van Le ◽  
Don Viet Nguyen ◽  
Quang Vu Nguyen ◽  
Bunmi Sherifat Malau-Aduli ◽  
Peter David Nichols ◽  
...  

Agronomy ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 941
Author(s):  
Ewa Szpunar-Krok ◽  
Anna Wondołowska-Grabowska ◽  
Dorota Bobrecka-Jamro ◽  
Marta Jańczak-Pieniążek ◽  
Andrzej Kotecki ◽  
...  

Soybean is a valuable protein and oilseed crop ranked among the most significant of the major crops. Field experiments were carried out in 2016–2019 in South-East Poland. The influence of soybean cultivars (Aldana, Annushka), nitrogen fertilizer (0, 30, 60 kg∙ha−1 N) and inoculation with B. japonicum (control, HiStick® Soy, Nitragina) on the content of fatty acids (FA) in soybean seeds was investigated in a three-factorial experiment. This study confirms the genetic determinants of fatty acid composition in soybean seeds and their differential accumulation levels for C16:0, C16:1, C18:1n9, C18:2, C18:3, and C20:0 as well saturated (SFA), monounsaturated (MUFA), and polyunsaturated (PUFA) fatty acids. Increasing the rate from 30 to 60 kg ha−1 N did not produce the expected changes, suggesting the use of only a “starter” rate of 30 kg ha−1 N. Inoculation of soybean seeds with a strain of Bradyrhizobium japonicum (HiStick® Soy, BASF, Littlehampton, UK and Nitragina, Institute of Soil Science and Plant Cultivation–State Research Institute, Puławy, Poland) is recommended as it will cause a decrease in SFA and C16:0 acid levels. This is considered nutritionally beneficial as its contribution to total fatty acids determines the hypercholesterolemic index, and it is the third most accumulated fatty acid in soybean seeds. The interaction of cultivars and inoculation formulation on fatty acid content of soybean seeds was demonstrated. An increase in the value of C16:0 content resulted in a decrease in the accumulation of C18:1, C18:2, and C18:3 acids. The content of each decreased by almost one unit for every 1% increase in C16:0 content. The dominant effect of weather conditions on the FA profile and C18:2n6/C18:3n3 ratio was demonstrated. This suggests a need for further evaluation of the genetic progress of soybean cultivars with respect to fatty acid composition and content under varying habitat conditions.


2000 ◽  
Vol 70 (1) ◽  
pp. 51-61 ◽  
Author(s):  
L. O. W. McClintont ◽  
A. F. Carson

AbstractThis study investigated the efficiency of growth and the carcass characteristics of 24 Greyface (Border Leicester × Scottish Blackface), 24 Texel (12 purebred and 12 Texel × Texel-Greyface) and 24 Rouge (12 purebred and 12 Rouge × Rouge-Greyface) lambs finished on the same level of feeding. The efficiency of live-weight gain (kg/MJ) was higher in Greyface compared with Texel lambs (P< 0·01). The efficiency of empty body-weight gain (kg/MJ) was higher in Greyface (P< 0·01) and Rouge (P< 0·05) compared with Texel lambs. The efficiency of carcass gains (kg/MJ) tended to be higher in Greyface and Rouge compared with Texel lambs (P= 0·07). The efficiency of non-carcass component gains (kg/MJ) was also higher in Greyface compared with Texel lambs (P0·05). Carcass water, protein, lipid and ash gains did not vary significantly between the genotypes, however carcass energy gain tended to be higher in Greyface and Rouge compared with Texel lambs (P= 0·08). The relative proportions of water, protein, lipid and ash in carcass gains did not vary significantly between the genotypes. At the end of the experiment carcass water content was higher in Texel compared with Greyface lambs (P< 0·05) and carcass ash content was lower in Texel compared with Greyface (P< 0·01) and Rouge (P< 0·05) lambs. The concentration of saturated fatty acids was higher in Greyface compared with Rouge lambs (P< 0·001) and higher in Rouge compared with Texel lambs (P< 0·05). Monounsaturated fatty acid concentrations were higher in Rouge compared with Greyface lambs (P< 0·05) and higher in Texel compared with Rouge lambs (P< 0·001). Polyunsaturated fatty acid concentrations were higher in Rouge and Texel compared with Greyface lambs (P< 0·01). The ratio of n-6:n-3 fatty acids was lower in Rouge compared with Greyface lambs (P< 0·05).The efficiency of empty body gain was higher in male compared with female lambs (P< 0·05). Carcass water (P< 0·01) and protein (P< 0·05) gains were higher in male lambs. At the end of the experiment male carcasses contained a higher content of water (P< 0·05), protein (P< 0·01) and ash (P= 0·07), and a lower lipid (P< 0·05) and energy (P< 0·001) content. Carcass lipids from male lambs contained a higher concentration of polyunsaturated fatty acids (P< 0·001) and tended to contain a lower concentration of saturated fatty acids (P = 0·06).


2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Zhouchen Ye ◽  
Jing Yu ◽  
Wuping Yan ◽  
Junfeng Zhang ◽  
Dongmei Yang ◽  
...  

AbstractCamellia oleifera (C. oleifera) is one of the four major woody oil-bearing crops in the world and has relatively high ecological, economic, and medicinal value. Its seeds undergo a series of complex physiological and biochemical changes during ripening, which is mainly manifested as the accumulation and transformation of certain metabolites closely related to oil quality, especially flavonoids and fatty acids. To obtain new insights into the underlying molecular mechanisms, a parallel analysis of the transcriptome and proteome profiles of C. oleifera seeds at different maturity levels was conducted using RNA sequencing (RNA-seq) and isobaric tags for relative and absolute quantification (iTRAQ) complemented with gas chromatography-mass spectrometry (GC-MS) data. A total of 16,530 transcripts and 1228 proteins were recognized with significant differential abundances in pairwise comparisons of samples at various developmental stages. Among these, 317 were coexpressed with a poor correlation, and most were involved in metabolic processes, including fatty acid metabolism, α-linolenic acid metabolism, and glutathione metabolism. In addition, the content of total flavonoids decreased gradually with seed maturity, and the levels of fatty acids generally peaked at the fat accumulation stage; these results basically agreed with the regulation patterns of genes or proteins in the corresponding pathways. The expression levels of proteins annotated as upstream candidates of phenylalanine ammonia-lyase (PAL) and chalcone synthase (CHS) as well as their cognate transcripts were positively correlated with the variation in the flavonoid content, while shikimate O-hydroxycinnamoyltransferase (HCT)-encoding genes had the opposite pattern. The increase in the abundance of proteins and mRNAs corresponding to alcohol dehydrogenase (ADH) was associated with a reduction in linoleic acid synthesis. Using weighted gene coexpression network analysis (WGCNA), we further identified six unique modules related to flavonoid, oil, and fatty acid anabolism that contained hub genes or proteins similar to transcription factors (TFs), such as MADS intervening keratin-like and C-terminal (MIKC_MADS), type-B authentic response regulator (ARR-B), and basic helix-loop-helix (bHLH). Finally, based on the known metabolic pathways and WGCNA combined with the correlation analysis, five coexpressed transcripts and proteins composed of cinnamyl-alcohol dehydrogenases (CADs), caffeic acid 3-O-methyltransferase (COMT), flavonol synthase (FLS), and 4-coumarate: CoA ligase (4CL) were screened out. With this exploratory multiomics dataset, our results presented a dynamic picture regarding the maturation process of C. oleifera seeds on Hainan Island, not only revealing the temporal specific expression of key candidate genes and proteins but also providing a scientific basis for the genetic improvement of this tree species.


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