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| Confirmation of introduced genes from PfFAD3-1 transformants (T 0 ) using PCR. Genomic DNA was extracted from Pb-con:PfFAD3-1 (A), Pphas : PfFAD3-1 (B), and P35S:PfFAD3-1 (C) T 0 transgenic leaf tissues. PfFAD3-1, PfFAD3-1 gene; Bar, Bar gene; Pb-con, b-conglycinin promoter; Pphas, phaseolin promoter; P35S, CaMV 35S promoter; NT, non-transgenic plant; #1-#21 (part A), Pb-con:PfFAD3-1 transgenic lines (T 0 ); #1-#7 (part B), Pphas : PfFAD3-1 transgenic lines (T 0 ); #1-#17 (part C), P35S:PfFAD3-1 transgenic lines (T 0 ).
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Soybean is a major crop that is used as a source of vegetable oil for human use. To develop transgenic soybean with high α-linolenic acid (ALA; 18:3) content, the FAD3-1 gene isolated from lesquerella (Physaria fendleri) was used to construct vectors with two different seed-specific promoters, soybean β-conglycinin (Pβ-con) and kidney bean phaseoli...
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... tissues from the 21 Pb-con:PfFAD3-1, 7 Pphas : PfFAD3-1, and 17 P35S:PfFAD3-1 transgenic plants (T 0 ) were used to confirm the integration of the transgene with PCR using PfFAD3-1 and Bar primers that amplified 1,146 and 548 bp DNA fragments, respectively. In addition, the DNA regions of the b-conglycinin, phaseolin, and CaMV 35S promoters were amplified as 548, 1,543, and 953 bp fragments, respectively ( Figure 2). The results of the PCR analysis for the 21 Pb-con: PfFAD3-1 transgenic lines confirmed the insertion of the transgene and b-conglycinin promoter sequence in all lines except line #15 (missing a region of the PfFAD3-1 gene) ( Figure 2A). ...
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... addition, the DNA regions of the b-conglycinin, phaseolin, and CaMV 35S promoters were amplified as 548, 1,543, and 953 bp fragments, respectively ( Figure 2). The results of the PCR analysis for the 21 Pb-con: PfFAD3-1 transgenic lines confirmed the insertion of the transgene and b-conglycinin promoter sequence in all lines except line #15 (missing a region of the PfFAD3-1 gene) ( Figure 2A). All seven putative Pphas : PfFAD3-1 transgenic plants successfully represented the transgene and phaseolin promoter ( Figure 2B). ...
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... results of the PCR analysis for the 21 Pb-con: PfFAD3-1 transgenic lines confirmed the insertion of the transgene and b-conglycinin promoter sequence in all lines except line #15 (missing a region of the PfFAD3-1 gene) ( Figure 2A). All seven putative Pphas : PfFAD3-1 transgenic plants successfully represented the transgene and phaseolin promoter ( Figure 2B). Among the 17 P35S:PfFAD3-1 transgenic lines, only line #7 was missing regions of both the PfFAD3-1 gene and CaMV 35S promoter ( Figure 2C). ...
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... seven putative Pphas : PfFAD3-1 transgenic plants successfully represented the transgene and phaseolin promoter ( Figure 2B). Among the 17 P35S:PfFAD3-1 transgenic lines, only line #7 was missing regions of both the PfFAD3-1 gene and CaMV 35S promoter ( Figure 2C). The selectable marker (Bar gene) was introduced in all PfFAD3-1-transformed soybean plants. ...
Citations
... Oleic acid is a monounsaturated fatty acid that promotes cardiac health by lowering low-density lipoprotein (LDL) cholesterol levels (Dhaka et al. 2011). Soybean oil also contains smaller amounts of alpha-linolenic acid (omega-3), another essential fatty acid known for its potential anti-inflammatory and cardiovascular benefits (Yeom et al. 2020). The balanced ratio of omega-3 to omega-6 fatty acids in soybeans contributes to its reputation as a healthy heart oil (Wijendran and Hayes 2004). ...
Owing to its high nutritional content of protein, oil, fatty acids, and sugars, soybean (Glycine max L.), one of the most significant legume crops, is utilized worldwide as food, feed, and fuel for daily life applications. The seeds, leaves, branches, roots, and pods of soybean contain essential bioactive compounds, including flavonoids, isoflavonoids, and other specialized metabolites, that play important roles in plant growth, development, and stress responses. In recent years, significant progress has been made in increasing soybean production. Therefore, here, we summarize the most recent breakthroughs in metabolite profiling and bioactive compound identification in soybeans to inform future digital breeding approaches. In addition to classical metabolite investigations, the discovery of metabolites involved in the dehydration response was made through a recent study that examined the regulatory network of metabolites and plant hormone genes. This review aimed to provide a metabolic perspective on soybeans that will benefit soybean production. The findings of this review will facilitate the development of novel soybean cultivars containing highly valuable metabolites for digital breeding.
... Previous studies reported that the desaturase genes play an important role in plant growth and development. They are used to synthesize unsaturated fatty acids in many planta, such as rice [31], walnuts [54], rape [55], and soybeans [56]. And, these desaturase genes, especially FAD2 and FAD3, also showed a positive response to abiotic stresses such as cold in rice, wheat, or olives [57][58][59][60]. ...
Rice is an important crop in the word, and fat is one of the main important nutrient components of rice. The lipid content and fatty acid composition of grains significantly influences the quality of rice. In this study, 94 homozygous recombination inbred lines (RILs) were developed and the crude fat content of them displayed a normal distribution ranging from 0.44% to 2.62%. Based on their taste quality, a positive association between fat content and eating quality was revealed. Then, two lines (FH and FL) were selected with similar agronomic characteristics and different lipid content and taste quality for RNA sequencing analysis, and a total of 619 differentiable expressed genes were detected, primarily enriched in metabolic pathways such as starch and sucrose metabolism, fatty acid metabolism, and amino acid metabolism. The expression of two genes related to fatty acid synthesis and elongation was significantly up-regulated, while the expression of three genes related to fatty acid degradation was significantly down-regulated in FH grains. By using liquid chromatography, the relative levels of palmitic acid and oleic acid were discovered significantly higher in FH grains. Additionally, the comparative genomic analysis was conducted to visualize genomic differences of five genes. Ultimately, two genes (Os07g0417200 and Os12g0102100) were selected to be the key gene to affect the lipid metabolism, especially for the synthesis of unsaturated fatty acids, significantly changing the eating quality of rice. These results provide a theoretical basis for improving the taste quality of rice.
... ALA, conversely, is less common: it is contained in the chloroplasts of green leafy vegetables and in walnuts, seeds of chia, flax, perilla and rape [3]. Other important sources of ALA are hemp seed oil and soybean oil (ALA content about 17% and 8%, respectively) [4,5]. However, despite various easily accessible soybean-derived products have an unchanged content of FAs (e.g., tempeh, tofu, etc.), others have a reduced content of FAs (e.g., soy milk, textured vegetable proteins, etc.) compared to the original source [6,7]. ...
Massive changes have occurred in our diet. A growing consumption of vegetal oils rich in omega-6 (ω-6) and a depletion of omega-3 (ω-3) fatty acids (FAs) in our food has led to an imbalance between ω-3 and ω-6. In particular, eicosapentaenoic (EPA)/arachidonic acid (AA) ratio seems to be an indicator of this derangement, whose reduction is associated to the development of metabolic diseases, such as diabetes mellitus. Our aim was therefore to investigate the literature on the effects of ω-3 and ω-6 FAs on glucose metabolism. We discussed emerging evidence from pre-clinical studies and from clinical trials. Notably, conflicting results emerged. Source of ω-3, sample size, ethnicity, study duration and food cooking method may be responsible for the lack of univocal results. High EPA/AA ratio seems to be a promising indicator of better glycemic control and reduced inflammation. On the other hand, linoleic acid (LA) appears to be also associated to a minor incidence of type 2 diabetes mellitus, although it is still not clear if the outcome is related to a reduced production of AA or to its intrinsic effect. More data derived from multicenter, prospective randomized clinical trials are needed.
... Attempts have been made to engineer ALA using seed-or endosperm-specific expression of FAD3 in plants with high-LA precursor, but most studies were performed not on oil crops (Liu et al. 2012). For oil crops, there are only four reports on ALA engineering in soybean and cottonseed, by over-expressing bifunctional Δ12/ω3 fatty acid desaturase from fungus Fusarium moniliforme (Damude et al. 2006), FAD3 from Arabidopsis thaliana (Eckert et al. 2006), FAD3-1 from Lesquerella (Yeom et al. 2020) and ...
α-Linolenic acid (ALA, 18:3Δ9,12,15) is an essential fatty acid for human, since it is the precursor for the biosynthesis of omega-3 long-chain polyunsaturated fatty acids (LC-PUFA). Modern people generally suffer from deficiency of ALA, since most staple food oils are low or lack in ALA content. Biotechnological enrichment of ALA in staple oil crops is a promising strategy. Among known oil crops, chia (Salvia hispanica) has the highest ALA content in its seed oil. In this study, the FAD2 and FAD3 genes from chia were engineered into a staple oil crop, oilseed rape (Brassica napus), via Agrobaterium tumefaciens-mediated transformation of their LP4-2A fusion gene construct driven by the seed-specific promoter PNapA. In seeds of T0, T1 and T2 lines, the average ALA contents were 20.86, 23.54 and 24.92%, which were 2.21, 2.68 and 3.03 folds of the non-transformed controls (9.42, 8.78 and 8.22%), respectively. The highest ALA levels of seeds of T0, T1 and T2 plants, were 38.41, 35.98 and 39.19% respectively, which were 4.10-4.77 folds of the respective controls. FA-pathway enzyme genes BnACCD, BnFATA, BnSAD, BnSCD, BnDGAT1, BnDGAT2 and BnDGAT3 as well as positive regulatory genes BnWRI1, BnLEC1, BnL1L, BnLEC2, BnABI3, BnbZIP67 and BnMYB96 were all significantly up-regulated. In contrast, BnTT1, BnTT2, BnTT8, BnTT16, BnTTG1 and BnTTG2, encoding negative regulators of oil accumulation but positive regulators of secondary metabolism, were all significantly down-regulated. These mean that foreign ShFAD2-ShFAD3 fusion gene directly and indirectly remodeled both positive and negative loci of the whole FA-related network in transgenic B. napus seeds.
... 12−14 A few cases on engineering ALA were performed mainly on nonoil crops, soybean and cottonseed. 15 Endosperm-specific expression of soybean FAD3-1 and rice FAD3 genes in rice increased the seed ALA content from 12.83 to 46.40%, compared with the control with 1.68%. 3 However, engineered ALA production in rice and tobacco using the overexpression of endogenous or heterogeneous FAD3/FAD7 gene achieved just a slight increase of the ALA level in seeds. ...
Modern people generally suffer from α-linolenic acid (ALA) deficiency, since most staple food oils are low in ALA content. Thus, the enhancement of ALA in staple oil crops is of importance. In this study, the FAD2 and FAD3 coding regions from the ALA-king species Perilla frutescens were fused using a newly designed double linker LP4-2A, driven by a seed-specific promoter PNAP, and engineered into a rapeseed elite cultivar ZS10 with canola quality background. The mean ALA content in the seed oil of PNAP:PfFAD2-PfFAD3 (N23) T5 lines was 3.34-fold that of the control (32.08 vs 9.59%), with the best line being up to 37.47%. There are no significant side effects of the engineered constructs on the background traits including oil content. In fatty acid biosynthesis pathways, the expression levels of structural genes as well as regulatory genes were significantly upregulated in N23 lines. On the other hand, the expression levels of genes encoding the positive regulators of flavonoid-proanthocyanidin biosynthesis but negative regulators of oil accumulation were significantly downregulated. Surprisingly, the ALA level in PfFAD2-PfFAD3 transgenic rapeseed lines driven by the constitutive promoter PD35S was not increased or even showed a slight decrease due to the lower level of foreign gene expression and downregulation of the endogenous orthologous genes BnFAD2 and BnFAD3.
... Physaria fendleri FAD3-1 (PfFAD3-1) was used to increase ALA levels in camelina seeds. PfFAD3-1 was chosen because soybean has a low 18:3 content; however, the expression of PfFAD3-1 in soybean significantly increased the 18:3 content by up to 52.4% 14,25,26 . P. fendleri is a plant species belonging to the Brassicaceae family, with oil containing 60% lesquerolic acid, which is a hydroxy fatty acid 27,28 . ...
... This similarity in the increase of 18:3 and decrease of 18:2 suggests that the conversion of 18:2 FA to 18:3 FA in camelina by PfFAD3-1 is both specific and efficient. In a previous study, when PfFAD3-1 was transformed into soybeans under the control of the seed-specific promoter, β-conglycinin, the ALA content increased by up to 52% in T1 seeds and up to 42% in T2 seeds 26 . In soybean, PfFAD3-1 expression increased ALA content under the control of the phaseolin promoter and 35S promoter; however, the increase in ALA content was the greatest under the control of β-conglycinin 26 . ...
... In a previous study, when PfFAD3-1 was transformed into soybeans under the control of the seed-specific promoter, β-conglycinin, the ALA content increased by up to 52% in T1 seeds and up to 42% in T2 seeds 26 . In soybean, PfFAD3-1 expression increased ALA content under the control of the phaseolin promoter and 35S promoter; however, the increase in ALA content was the greatest under the control of β-conglycinin 26 . PfFAD3-1 expression in soybeans did not result in significantly different oil content from that of the WT; however, it increased both seed size and weight 26 . ...
Camelina (Camelina sativa) is an oil crop with a short growing period, resistance to drought and cold, low fertilizer requirements, and can be transformed using floral dipping. Seeds have a high content of polyunsaturated fatty acids, especially ɑ-linolenic acid (ALA), at 32–38%. ALA is an omega-3 fatty acid that is a substrate for eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in the human body. In this study, ALA content was further enhanced by the seed-specific expression of Physaria fendleri FAD3-1 (PfFAD3-1) in camelina. The ALA content increased up to 48% in T2 seeds and 50% in T3 seeds. Additionally, size of the seeds increased. The expression of fatty acid metabolism-related genes in PfFAD3-1 OE transgenic lines was different from that in the wild type, where the expression of CsFAD2 decreased and CsFAD3 increased. In summary, we developed a high omega-3 fatty acid-containing camelina with up to 50% ALA content by introducing PfFAD3-1. This line can be used for genetic engineering to obtain EPA and DHA from seeds.
... High oleic acid and low linolenic acid soybeans were produced by incorporating two loci for low linolenic acid including mutant GmFAD3A and GmFAD3C (Lakhssassi et al., 2017. In contrast to this, a heterologous gene PfFAD3-1 from Physaria fendleri was expressed in soybean seeds to produce oil reach in ω-3ALA (Yeom et al., 2020) keeping in mind the importance of ALA in producing eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in the human body. Further, the soybean oil enriched in EPA and DHA could be used in manufacturing food and feed products. ...
... High oleic acid and low linolenic acid soybeans were produced by incorporating two loci for low linolenic acid including mutant GmFAD3A and GmFAD3C (Lakhssassi et al., 2017. In contrast to this, a heterologous gene PfFAD3-1 from Physaria fendleri was expressed in soybean seeds to produce oil reach in ω-3ALA (Yeom et al., 2020) keeping in mind the importance of ALA in producing eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in the human body. Further, the soybean oil enriched in EPA and DHA could be used in manufacturing food and feed products. ...
Several million hectares of economically cultivated transgenic crops are currently available. There have been field trials of transgenics from at least 200 species, oil plants (including both annual and perennial) belonging to different plant families. Crops that produce oil seeds are crucial to the agricultural economy. They have a significant role in industrial uses and the human diet, but they are also becoming more significant as a fossil fuel replacement for supplying energy. A number of significant developments in genetic engineering and the identification of gene targets for increasing seed oil content in oilseed crops have occurred during the past 20 years. These developments will help the successful development of new generation high yielding oil crops. A significant portion of the most recent data comes from the examination of patent databases, an excellent source of data on commercial priorities. The timeline for the introduction of these transgenes into breeding populations and their eventual release as novel varieties is also covered in the review.
... For example, a commercial soybean oil with high oleate levels may be superior to conventional oil in terms of oxidative stability [1]. Similarly, increasing the level of omega-3 linolenic and lowering the content of omega-6 linoleic in soybean meet the target for the prevention of obesity and cardiovascular diseases [8]. The demand of vegetable oils is growing rapidly, and the current annual consumption of 200 million metric tons may be doubled by 2050, which will impose a great burden on the limited availability of arable land [9]. ...
... The conversion of 18:2 to 18:3 is catalyzed by omega-3 fatty acid desaturase (FAD3) in the ER [48]. 18:3 is essential for the prevention of cardiovascular and cerebrovascular diseases in humans and animals [8]. In the soybean genome, four genes encoding FAD3 desaturases are designated as GmFAD3A (GmFAD3-1b), GmFAD3B (GmFAD3-1a), GmFAD3C (GmFAD3-2a) and GmFAD3E [32,70]. ...
... To optimize the omega-6/omega-3 ratio in soybean or other oil crops, the introduction of exogenous FAD3s from high-18:3-TAG plants, such as Physaria fendleri or peony trees, provides approaches to substantially increase linolenic acid proportions [74,75]. Seed-specific expression of PfFAD3-1 (driven by the promoter of soybean β-conglycinin) resulted in 42% linolenic acid content in soybean seeds, while only about 10% accumulated using a 35S constitutive promoter construct [8]. PfFAD3 lines showed a significant improvement in seed yield and seed size, probably as a consequence of increasing adaptability to stress tolerance [8]. ...
Soybean is a major oil crop and is also a dominant source of nutritional protein. The 20% seed oil content (SOC) of soybean is much lower than that in most oil crops and the fatty acid composition of its native oil cannot meet the specifications for some applications in the food and industrial sectors. Considerable effort has been expended on soybean bioengineering to tailor fatty acid profiles and improve SOC. Although significant advancements have been made, such as the creation of high-oleic acid soybean oil and high-SOC soybean, those genetic modifications have some negative impacts on soybean production, for instance, impaired germination or low protein content. In this review, we focus on recent advances in the bioengineering of soybean oil and its effects on agronomic traits.
... Therefore, the cv. Kwangan was broadly cultivated in Korea and used as a well-established parental plant to develop the Biotech soybean (Kim et al., 2012;Yeom et al., 2020). However, the molecular response of cv. ...
Soybean ( Glycine max (L) Merr.) provides plant-derived proteins, soy vegetable oils, and various beneficial metabolites to humans and livestock. The importance of soybean is highly underlined, especially when carbon-negative sustainable agriculture is noticeable. However, many diseases by pests and pathogens threaten sustainable soybean production. Therefore, understanding molecular interaction between diverse cultivated varieties and pathogens is essential to developing disease-resistant soybean plants. Here, we established a pathosystem of the Korean domestic cultivar Kwangan against Pseudomonas syringae pv. syringae B728a. This bacterial strain caused apparent disease symptoms and grew well in trifoliate leaves of soybean plants. To examine the disease susceptibility of the cultivar, we analyzed transcriptional changes in soybean leaves on day 5 after P. syringae pv. syringae B728a infection. About 8,900 and 7,780 differentially expressed genes (DEGs) were identified in this study, and significant proportions of DEGs were engaged in various primary and secondary metabolisms. On the other hand, soybean orthologs to well-known plant immune-related genes, especially in plant hormone signal transduction, mitogen-activated protein kinase signaling, and plant-pathogen interaction, were mainly reduced in transcript levels at 5 days post inoculation. These findings present the feature of the compatible interaction between cultivar Kwangan and P. syringae pv. syringae B728a, as a hemibiotroph, at the late infection phase. Collectively, we propose that P. syringae pv. syringae B728a successfully inhibits plant immune response in susceptible plants and deregulates host metabolic processes for their colonization and proliferation, whereas host plants employ diverse metabolites to protect themselves against infection with the hemibiotrophic pathogen at the late infection phase.
