• Proceedings of the Korean Society of Crop Science Conference
• /
• 2022.10a
• /
• pp.24-24
• /
• 2022

This present study was to identify a novel candidate gene that contribute to the elevated α-linolenic acid (ALA, ω-3) concentration in PE2166 from mutagenesis of Pungsannamul. Major loci qALA5_1 and qALA5_2 were detected on chromosome 5 of soybean through quantitative trait loci mapping analyses of recombinant inbred lines. With next generation sequencing of parental lines and Pungsannamul, and recombinant analyses, a potential gene, Glyma. 05g221500 (HD) controlling elevated ALA concentration was identified. HD is a homeodomain-like transcriptional regulator that may regulate the expression level of microsomal ω-3 fatty acid desaturase (FAD3) genes responsible for the conversion of linoleic acid into ALA in the fatty acid biosynthetic pathway. In addition, we hypothesized that combination of mutant alleles, HD and either of microsomal delta-12 fatty acid desaturase 2-1 (FAD2-1\ could reduce the ω-6/ω-3 ratio. In populations where HD, and FAD2-1A and FAD2-1B genes were segregated, combination of a hd allele from PE2166 and either of the variant FAD2-1 alleles were sufficient to reduce the ω-6/ω-3 ratio in seeds.

• Korean Journal of Breeding Science
• /
• v.50 no.4
• /
• pp.463-471
• /
• 2018

Perilla is an oilseed crop cultivated in Korea since ancient times. Due to the high ${\alpha}-linolenic$ acid content in perilla, perilla seed oil can easily become rancid. ${\alpha}-Linolenic$ acid is synthesized by two enzymes, endoplasmic reticulum-localized ${\Delta}15$ desaturase (FAD3) and chloroplast-localized ${\Delta}15$ desaturase (FAD7) in vivo. In order to lower the ${\alpha}-linolenic$ acid content of the seed oil without disturbing plant growth, we tried to suppress the expression of only the FAD3 gene using RNA interference, whilst maintaining the expression of the FAD7 gene. Seventeen transgenic plants with herbicide ($Basta^{TM}$) resistance were obtained by Agrobacterium-mediated transformation using hypocotyls of perilla plants. The transgenic plants were firstly confirmed by treatment with 0.3% (v/v) $Basta^{TM}$ herbicide, and the expression of FAD3 was measured by Northern blot analysis. The ${\alpha}-linolenic$ acid content was 10-20%, 30-40%, and 60% in two, seven, and three of the twelve $T_1$ transgenic perilla plants which had enough seeds to be analyzed for fatty acid composition, respectively. Analysis of the fatty acid composition of $T_2$ progeny seeds from $T_1$ plants with the lowest ${\alpha}-linolenic$ acid content showed that the homozygous lines had 6-10% ${\alpha}-linolenic$ acid content and the heterozygous lines had 20-26% ${\alpha}-linolenic$ acid content. It is expected that the reduction in ${\alpha}-linolenic$ acid content in perilla seed oil will prevent rancidity and can be utilized for the production of high-value functional ingredients such as high ${\gamma}-linolenic$ acid.

• KSBB Journal
• /
• v.20 no.2 s.91
• /
• pp.93-99
• /
• 2005

Polyunsaturated fatty acids, that is PUFAs, are important constituents of membranes particularly found in the retina and central nervous system. In microorganism-based PUFAs biosynthesis, the genus Thraustochytrids is well evaluated for their potential as a promising candidate in the practical production of PUFAs, such as AA and DHA. In this study, we attempted to optimize a method of total nucleic acid extraction from this microorganism as a preliminary experiment. Using the extracted nucleic acid and degenerated primers for direct PCR, we isolated a $\Delta5$ desaturase gene that contained 1320-nucleotide and encoded 439 amino acids. This gene exhibited an expected function, when expressed in P. pastoris in the presence of appropriate exogenous substrate, as an evidence for $\Delta5$ desaturase activity (conversion of DGLA to AA). These results and information could provide a basis for the construction of engineered strains suitable for the practical production of PUFAs.

• Asian-Australasian Journal of Animal Sciences
• /
• v.32 no.1
• /
• pp.137-143
• /
• 2019

Objective: The aim of this study was to compare male and female geese of two contrasting genotypes in terms of fatty acid composition, indexes related to human health, lipid metabolism and oxidative stability of the meat. Methods: The experiment was carried out on total of 120 geese of two different genotypes; the native breed Czech goose (CG) and commercial hybrid Novohradska goose (NG). One-d-old goslings were divided into 4 groups according to genotype and sex, and 8 birds from each group were slaughtered at 8 weeks of age. Results: The effects of the interactions between genotype and sex were observed on growth performance and carcass traits. Final body weight (p<0.001), daily weight gain (p<0.001), daily feed intake (p<0.001), slaughter weight (p<0.001), and cold carcass weight (p<0.001) were highest in NG males and lowest in CG females. The meat fatty acid composition results showed effects of both genotype and sex on the total n-6 and the total polyunsaturated fatty acid (PUFA) content, as well as the PUFA n-6/PUFA n-3 ratio. Regarding genotype, the total n-6, the total PUFA content and the PUFA n-6/PUFA n-3 ratio were higher in CG, and higher values were found in females. In terms of the lipid metabolism, ${\Delta}^5-{\Delta}^6$ desaturase (p = 0.006) was higher in males. The meat oxidative stability results revealed an interaction between genotype, sex and storage time (p<0.001). The highest (13.85 mg/kg) malondialdehyde content was measured in the meat of CG females after 5 days of storage and was presumably related to a higher PUFA content. Conclusion: NG had a relatively higher growth rate and meat oxidative stability, whereas the advantage of CG meat is its favourable fatty acid profile characterized by a higher PUFA content.

• Journal of Animal Science and Technology
• /
• v.55 no.2
• /
• pp.147-153
• /
• 2013

Lipid metabolism in mature male mice may be different from immature male mice, but the relationship of lipid metabolism, especially n-6 fatty acid metabolism, and sexual maturation is not clearly established. This study was carried out to elucidate whether sexual maturation may affect the metabolism of functional n-6 fatty acids of lipid components by investigating the composition of fatty acids in the longissimus muscle tissues of mature and immature male mice with GC and analyzing the expression of genes and proteins for synthesis of n-6 fatty acids with real-time PCR and western blotting, respectively. Mature male mice showed significantly higher testosterone level in the sera. Similarly, n-6 fatty acids, levels of linoleic acid (LA 18:2n-6) and total n-6 PUFA (Polyunsaturated fatty acids) were increased, but the levels of ${\gamma}$-linolenic acid (GLA; 18:3n-6), dihomo-${\gamma}$-linolenic acid (DGLA; 20:3n-6) and arachidonic acid (AA; 20:4 n-6) were decreased in the mature male mice. mRNA levels of ${\Delta}5$-desaturase (FASD1) and elongase (ELOVL5) genes related to n-6 fatty acid metabolism increased. However, the level of FADS1 protein only increased in mature male mice. In conclusion, this study suggested that sexual maturation of male mice affected n-6 fatty acid metabolism by stimulating the expression of enzyme FADS1 of n-6 PUFA metabolism.

• Development and Reproduction
• /
• v.22 no.4
• /
• pp.297-307
• /
• 2018

The objective of this study was to evaluate the effects of alpha-linolenic acid (ALA) during in vitro maturation (IVM) on cumulus expansion, nuclear maturation, fertilization capacity and subsequent development in porcine oocytes. The oocytes were incubated with 0, 25, 50, and $100{\mu}M$ ALA. Cumulus expansion was measured at 22 h, and gene expresison and nuclear maturation were analyzed at 44 h after maturation. Then, mature oocytes with ALA were inseminated, and fertilization parameters and embryo development were evaluated. In results, both of cumulus expansion and nuclear maturation were increased in $50{\mu}M$ ALA groups compared to control groups (p<0.05). However, expression of gap junction protein alpha 1 (GJA1, cumulus expansion-related gene), delta-6 desaturase (FADS1, fatty acid metabolism-related gene), and delta-5 desaturase (FADS2) mRNA in cumulus cells were reduced by $50{\mu}M$ ALA treatment (p<0.05). Cleavage rate was enhanced in 25 and $50{\mu}M$ ALA groups (p<0.05), especially, treatment of $50{\mu}M$ ALA promoted early embryo develop to 4 and 8 cell stages (p<0.05). However, blastocyst formation and number of cells in blastocyst were not differ in 25 and $50{\mu}M$ ALA groups. Our findings show that ALA treatment during maturation could improve nuclear maturation, fertilization, and early embryo development through enhancing of cumulus expansion, however, fatty acid metabolism- and cumulus expansion-related genes were down-regulated. Therefore, addition of ALA during IVM of oocytes could improve fertilization and developmental competence, and further studies regarding with the mechanism of ALA metabolism are needed.

• Korean journal of applied entomology
• /
• v.56 no.1
• /
• pp.61-67
• /
• 2017

Two major sex pheromone components (Z-8-dodecenyl acetate and E-8-dodecenyl acetate) are known in the peach fruit moth, Grapholita molesta. From a putative biosynthetic pathway of these sex pheromone components, delta 10 desaturase ($10{\Delta}$ DES) has been proposed to play a crucial role in synthesizing a species-specific stereoisomer of the double bond. However, its molecular identity was not known. This study determined a putative desaturase (Gm-comp1575) as a $10{\Delta}$ DES candidate from G. molesta transcriptome constructed from the sex pheromone gland. Its open reading frame encodes 370 amino acid sequence with a predicted molecular weight at 43.2 kDa and isoelectric point at 8.77. It was predicted to have four transmembrane domains and six glycosylation sites at N-terminal or cytosolic domains. A phylogenetic analysis with its predicted amino acid sequence indicated that Gm-comp1575 is closely related with known $10{\Delta}$ DES genes of other insects. Gm-comp1575 transcript was detected in female adults at sex pheromone gland and other abdominal tissues. RNA interference of Gm-comp1575 significantly reduced attractiveness of virgin females in apple orchard compared to control females. These results suggest that Gm-comp1575 is associated with sex pheromone biosynthesis of G. molesta.

• Journal of Embryo Transfer
• /
• v.30 no.2
• /
• pp.83-89
• /
• 2015

The sexual maturation occurred by the changes of steroid hormones was known to sex-dependently and/or age-dependently regulate the lipid metabolism in various animal species. Our current study demonstrates that lipid and its functional fatty acids can be changed depending on the status of sexual maturation. Of the functional fatty acids, ${\gamma}$-linolenic acid (GLA; 18:3n-6) is an important factor for maintaining human health. The purpose of our study was to investigate the level of GLA in mice with different stages of sexual maturation. To this end, the longissimus muscle (LM) of immature (3-week-old) and mature (7-week-old) female mice was analysed for the fatty acid composition by gas chromatography. Furthermore, both gene and protein level of ${\Delta}6$ desaturase (FADS2) which is involved in GLA metabolism by real time PCR and Western blotting, respectively. Mature females showed greater (P<0.05) serum $17{\beta}$-estradiol (E2) level and LM GLA contents than immature group. The mRNA and protein levels of FADS2, which converts precursor linoleic acid into GLA, were higher (P<0.05) in mature female mice than in immature mice. In conclusion, these results show that sexual maturation of female mice induces GLA and FADS2 contents in LM.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.32 no.2
• /
• pp.269-277
• /
• 2003

Effects of soy protein concentrate (SPC) containing isoflavone and casein diets on plasma phospholipid (PLs)-fatty acid patterns were investigated in 7-& 40- wk old female rats. Diets containing 16% SPC (soy/young:SY, soy/old: SO) and casein (casein/young : CY, casein/ old: CO) supplemented with 0.5% cholesterol were fed for 4 wks. Fatty acid compositions of plasma PLs were determined by TLC and GLC. Compared to the dietary protein effects, age effects on serum lipids were more profound. The levels of total cholesterol (Chol.), triglyceride, HDL-Chol., (LDL+VLDL)-Chol. and atherogenic index (AI) were higher in older groups (OC & OS) than younger groups (YC &. YS). Soy groups had higher Ell)L-Chol. level and lower (LDL+ VLDL)-Chol. and AI, compared with casein groups. The compositions of C22:0, Cl8:1 $\omega$9 and sum of MUFA in plasma PLs were significantly higher in casein group (CY & CO) than soy group (SY & SO), but those of sum of SFA were higher in soy group. The compositions of C22:0, Cl8:1 $\omega$9, C22:1, Cl8:3$\omega$3 and C22:4$\omega$6 were higher and those of C22:6$\omega$3, sum of $\omega$3, Cl8: 2$\omega$6 C20:4$\omega$6, sum of $\omega$6 and sum of PUFA were lower in plasma PLs of younger rats. The average P/S and $\omega$3/$\omega$6 ratio in older group was higher. The $\Delta$-7 desaturation index (16:0⇒16:1$\omega$7) and $\Delta$-9 desaturation index (18:0⇒18:1$\omega$9) were lower in soy group than casein group, while $\Delta$-6 and $\Delta$-5 desaturation index were not affected by dietary protein. The $\Delta$-4 desaturation index (22:4$\omega$6⇒22:5$\omega$6) were higher and elongation index (20:4$\omega$6⇒22:4$\omega$6) were lower in older group. The ratio of the products of $\omega$3 fatty acid series (Cl8:3) was significantly higher in older group, which indicated that age affected the plasma PUFA metabolism. On the other hand, older rats had higher serum cholesterol level compared with younger rats. Taken together, these changes in fatty acid composition might cause minimal changes in tile membrane fluidity induced by the increase serum cholesterol level.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.33 no.10
• /
• pp.1626-1633
• /
• 2004

In recent, the roles of oleic acid, most abundant fatty acid in myelin, were investigated in relation to the brain functions. This study examined the effects of diets either with desirable ratios of $\omega$6/ $\omega$9 and P/M/S (mixed oil-fed group: MO) or with defficient in $\omega$3 series fatty acids (safflower oil-fed group: SO) on the oleic acid composition in RBC and brain synaptosomal, mitochondrial & microsomal phospholipids. The desirable fatty acid composition was computer-searched with different fats and oils to meet right ratios of both $\omega$6/ $\omega$3 and P/M/S. Diets were fed 3 weeks before conception and new-born pups were fed maternal milk from the same mothers and same diets until 9 wks of age. At 3 wks of age, the compositions of oleic acid in brain subcellular fractions and red blood cells were constantly remained whatever the composition of dietary fatty acids. But at 9 wks of age, the composition of oleic acid in synaptosome and mitochondria were significantly higher in MO group than SO group. The composition of oleic acid in milk was significantly higher in MO group than SO group, but in case of SO group, that of oleic acid was increased by 48%, in comparison with dietary fatty acid compositions. -9 desaturase index (18:0\longrightarrow8:1) of brain synaptosome was significantly higher in MO group than SO group at 3 and 9 weeks of ages, but that of brain microsome was higher in SO group than MO group at 9 wks of age. Taken together, the presences of oleic acid in the diet was important to maintain brain functions. The origins of oleic acid in brain may suggests two hypotheses; first, the central nervous system has priority for the uptake of oleic acid, and second the nervous system can synthesize all the oleic acid it needs, independently of its presence in the diet.