Alpha-linolenic acid is an important polyunsaturated fatty acid that exhibits anticancer, anti-inflammatory, and antioxidative effects. In this study, we investigated the protective effects of alpha-linolenic acid on the cell proliferation and differentiation of C2C12 cells under essential amino acid-deficient conditions. Different concentrations of alpha-linolenic acid and essential amino acids were added to the growth and differentiation media. The concentrations of 10 μM of alphalinolenic acid and 2% essential amino acid were chosen for subsequent experiments. Supplementation with alpha-linolenic acid and essential amino acids improved the proliferation and differentiation of C2C12 cells and significantly increased the mRNA levels of catalase, superoxide dismutase, B-cell lymphoma-2, and beclin-1 as well as the protein levels of PPARγ coactivator-1α compared to those in the controls. Moreover, supplementation with alpha-linolenic acid and essential amino acids reduced the levels of phosphorylated H2A.X variant histone, Bcl-2-associated X, p53, and light chain 3 during C2C12 cell proliferation, and increased the expression levels of myogenic factors 4 (myogenin) and 5 during C2C12 cell differentiation. Overall, we determined that alpha-linolenic acid and essential amino acids maintained the cell proliferation and differentiation of C2C12 cells via their anti-oxidative, anti-apoptotic, and anti-autophagic effects.

The aim of present study was to investigate regulatory mechanism of alpha-linolenic acid (ALA) during in vitro maturation (IVM) on nuclear and cytoplasmic maturation of porcine oocytes. Basically, immature cumulus-oocyte complexes (COCs) were incubated for 22 h in IVM-I to which hormone was added, and then further incubated for 22 h in IVM-II without hormone. As a result, relative cumulus expansion was increased at 22 h after IVM and it was enhanced by treatment of ALA compared with control group (p < 0.05). During IVM process within 22 h, cAMP level in oocytes was decreased at 6 h (p < 0.05) and it was recovered at 12 h in ALA-treated group, while oocytes in control group recovered cAMP level at 22 h. In cumulus cells, it was reduced in all time point (p < 0.05) and ALA did not affect. Treatment of ALA enhanced metaphase-I (MI) and MII population of oocytes compared with oocytes in control group at 22 and 44 h, respectively (p < 0.05). Intracellular GSH levels in ALA group was increased at 22 and 44 h after IVM (p < 0.05), whereas it was increased in control group at 44 h after IVM (p < 0.05). In particular, the GSH in ALA-treated oocytes during 22 h of IVM was higher than control group at 22 h (p < 0.05). Lipid amount in oocytes from ALA group was higher than control group (p < 0.05). Treatment of ALA did not influence to absorption of glucose from medium. Cleavage and blastocyst formation of ALA-treated oocytes were enhanced compared with control group (p < 0.05). These findings suggest that supplementation of ALA could improve oocyte maturation and development competence through increasing GSH synthesis, lipid storage, and regulation of cAMP accumulation during early 22 h of IVM, and these might be mediated by cumulus expansion.

This present study was conducted to investigate protective effect of discontinuous Percoll gradient containing alpha-linolenic acid (ALA) before freezing process on viability, acrosome damage, mitochondrial activity, and oxidative stress of frozen-thawed boar spermatozoa. The separation of spermatozoa by discontinuous Percoll gradient was performed by different concentration of Percoll solution (45/90%) containing ALA combined with bovine serum albumin (BSA), and collected sperm in each Percoll layer was cryopreserved. To evaluate viability, acrosome damage, mitochondrial activity, and reactive oxygen species (ROS) level of frozen-thawed sperm, flow cytometry was used. Morphological abnormalities were observed under light microscope. In results, viability of sperm from 90% Percoll layer was higher than control and 45% Percoll group (p < 0.05). Separated sperm in 90% Percoll layer had lower acrosome damage and morphological abnormalities than control as well as viability, whereas 45% Percoll group was higher (p < 0.05). Similar with acrosome damage and abnormalities, mitochondrial activity was slightly enhanced and the population of live sperm with high ROS level was decreased by 90% Percoll separation, however, there was no significant difference. Supplementation of 3 ng/mL ALA into Percoll solution increased sperm viability and decreased population of live sperm with high ROS compared to control (p < 0.05). In conclusion, discontinuous Percoll gradient before freezing process could improve efficiency of cryopreservation of boar sperm through selection of sperm with high freezing resistance, and supplement of ALA during Percoll gradient might contribute suppression of ROS generation via stabilizing of plasma membrane during cryopreservation.

Alpha-linolenic acid (ALA; n-3 18:3), a one of omega-3 fatty acid, is mainly contained in chloroplast of plant and ALA is an essential fatty acid, not synthesized in mammalian body, it must be supplied from foods. Polyspermy is especially high on in vitro fertilization (IVF) in pigs, which is a major obstacle to in vitro embryo production systems. In our previous study, when ALA was supplemented during in vitro maturation (IVM), the methaphase-II rate and gluthathione level was increased. The objective of this study was to evaluate the effects of alpha-linolenic acid (ALA) supplementation during IVM and subsequent of IVF in pigs. The cumulus-oocyte complexes (COCs) were submitted to IVM medium containing 0, 25, 50, and 100 μM ALA for 44 h. After 44 h of IVM, denuded oocytes were co-cultured with spermatozoa during 18 h. After 18 h of in vitro fertilization, oocyte were using aceto-orcein method, to evaluated penetration rate, monospermy (number of monospermy oocytes/total oocytes), and the IVF efficiency (number of monospermy/total penetrated oocytes). In results, 25 and 50 μM ALA groups were significantly increased on penetration rate compared with 100 μM ALA group (p<0.05). Similarly, monospermy rate were significantly increased 25 and 50 μM ALA groups than control group (p<0.05). IVF efficiency was no significant difference between control and ALA treatment groups. Our findings suggested that treatment of ALA supplementation during in vitro maturation (IVM) and subsequent of in vitro fertilization in pigs, ALA can increase IVF efficiency by effectively blocking polyspermy and increasing monospermy some mechanism in porcine oocytes. However, the study of mechanism by which ALA blocks polyspermy are needed, and this study suggests that ALA has a positive effect on in vitro production of porcine oocytes by decreasing polyspermy. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (Ministry of Education) (2016R1D1A1B03931746).

During the freezing and thawing process, fatty acids in the plasma membrane of sperm are released, which results in a functional damage of sperm. Sperm with functional loss due to cryo-damage result in a decrease in fertility. Previous studies have shown that the addition of one of the fatty acid alpha-linolenic (ALA) with carrier proteins improves the stability of plasma membrane and reduces the damage. In this experiment, we focused on the functional aspects of the plasma membrane of sperm and experimented with motility and morphology. For preparation of ALA-carrier protein complex, 3 ng/ml ALA was mixed with 0.7 μg/ml bovine serum albumin (BSA) or 14 ng/ml methyl-β-cyclodextrin (MBCD) in distilled water. The boar semen was purchased from GUMBO Company. Boar semen was cryo-preserved in 20% egg yolk freezing extender containing ALA, BSA, MBCD, ALA+BSA, ALA+MBCD. The frozen boar sperm was thawed at 37.5 ℃ for 45 sec in water-bath. The sperm motility and morphological abnormalities were evaluated under a phase-contrast microscope at 200 × magnification and randomly counts of 200 sperm each sample. In results, motility of frozen-thawed sperm was increased in all treatment groups. In particular, there has been significant improvement in ALA+BSA and ALA+MBCD treatment groups than control (p<0.05). However, there was no significant difference in ALA, BSA and MBCD treatment groups. Morphological normalities in frozen-thawed sperm was reduced in complex treatment groups (p<0.05). However, there was no significant difference in single treatment groups. In both motility and morphology characteristics, ALA+BSA and ALA+MBCD treatment group was higher than all treatment groups. In conclusion, the addition of ALA with carrier proteins during cryopreservation has a positive effect in its functional aspect. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (Ministry of Education) (2016R1D1A1B03931746).

Alpha-linolenic acid (ALA) is one of n-3 polyunsaturated fatty acids and found mainly in the chloroplasts. Many studies have been reported that intracellular reactive oxygen species (ROS) in mammalian oocytes were reduced by supplementation of ALA in in vitro maturation (IVM) medium. Based on these reports, we expected that ALA acts as an antioxidant during IVM of porcine oocytes. Therefore, the objective of this study was to investigate the antioxidant effect of ALA supplementation during IVM in porcine oocytes. The cumulus-oocyte complexes (COCs) were incubated in IVM medium containing 200 μM H2O2 or H2O2 with 50 μM ALA for 44 h. Nuclear maturation stage of oocytes was evaluated using aceto-orcein method. For measurement of oxidative stress state, intracellular ROS and glutathione (GSH) levels were measured using carboxy-DCFDA and cell tracker red, respectively. In results, oocytes in metaphase-II (MII) stage development was significantly reduced in H2O2 group compared to non-treated control group (61.84±1.42% and 80.00%, respectively; p<0.05) and it was slightly recovered by treatment of ALA (69.76±1.67%; p<0.05). The intracellular GSH levels was decreased in H2O2 groups compared with control groups, but it was enhanced by ALA treatment (p<0.05). On the contrary, H2O2 treatment increased intracellular ROS level in oocytes and H2O2-induced ROS was decreased by treatment of ALA (p<0.05). Our findings suggested that ALA treatment under oxidative stress condition improve oocyte maturation via elevated GSH and reduced ROS levels in oocytes. Therefore, these results suggest that ALA have an antioxidative ability and it could be used as antioxidant in in vitro production system of porcine embryo.

This study was conducted to evaluate effect of α-linolenic acid (ALA) and bovine serum albumin (BSA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved using freezing extender containing 3 ng/mL ALA and/or 20 μg/mL BSA. Cryopreserved boar sperms were thawed in 37°C water-bath for 45 sec to analysis. Viability, acrosome reaction, and mitochondrial intact were analyzed using flow cytometry. In results, viability of frozen-thawed boar sperm was significantly higher in only ALA+BSA supplement group than control group (p<0.05), whereas there was no difference either in ALA or BSA supplement. However, acrosome reacted sperm in both of live and all sperm population were significantly decreased in all treatment groups than control (p<0.05). Interestingly, mitochondrial intact of boar sperm was enhanced in ALA and ALA+BSA groups compared with control (p<0.05). In this study, we showed that supplementation of ALA and BSA in freezing extender enhanced the sperm viability, mitochondrial intact and decrease acrosomal membrane damage. In conclusion, our findings suggest that quality of frozen-thawed sperm in mammalians could improve by using of ALA and BSA.

This study investigated the effect of dietary lysine and gamma-linolenic acid(GLA) levels on growth performance, carcass traits, and meat quality in finishing pigs. Pigs were provided with feed containing two different levels of lysine(0.45% and 0.75%) with three different levels of gamma-linolenic acid(0.0, 0.3, and 0.6%). Average daily gain(ADG) was significantly lower (p<0.01) in pigs provided with the lower level of lysine. In contrast, feed/gain(p<0.01), diet cost/gain(p<0.05), and intramuscular fat(p<0.01) were all significantly higher in pigs fed the lower level of lysine. Similarly, meat color scores(CIE L*, a*, and b*) and cooking loss were significantly higher(p<0.01) in pigs fed the lower level of lysine, whereas shear force(kg/2.5 inch2)was not affected by dietary lysine. The addition of GLA had no significant effect on any of the parameters measured. The results indicate that providing pigs with 0.45% lysine in their diet may help to increase intramuscular fat content, allowing the industry to produce pork products that meet consumer needs in Korea.

The aim of this study was to evaluate effect of α-linolenic acid (ALA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved in 20% egg yolk freezing extender containing ALA (0, 3, 5, and 10 ng/mL) with 0.05% ethanol. The frozen-boar spermatozoa were thawed at 37.5°C for 45 sec in water-bath. The spermatozoa samples were evaluated the plasma membrane integrity, acrosome reaction, and mitochondrial integrity using flow cytometry. In results, population of live sperm with intact plasma membrane was significantly higher in control and 3 ng/mL ALA treatment group than ethanol group (p<0.05). In contract, dying sperms were higher in ethanol group than 3 ng/mL ALA treatment (p<0.05). Acrosomal membrane damage in all sperm population was reduced in 3 ng/mL ALA groups compared with ethanol treatment (p<0.05). However, acrosome damage in live sperm population was no significant difference among the all treatment groups. Mitochondrial integrity was not influenced by ALA treatments in both of live and all sperm population. In conclusion, this results show that supplement of ALA during the cryopreservation process could reduce the membrane damages including plasma and acrosomal membrane, whereas ALA did not influence to mitochondria in boar spermatozoa. Therefore, these results suggest that ALA can protect against the membrane damage derived cryo-stress, and cryopreservation efficiency of boar semen would be improved by use of ALA.

The sexual maturation occurred by the changes of steroid hormones was known to sex-dependently and/or agedependently 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, γ- 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 Δ6 desaturase (FADS2) which is involved in GLA metabolism by real time PCR and Western blotting, respectively. Mature females showed greater (P<0.05) serum 17β -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.

This study examined the effects of different levels of evening primrose oil(EPO) on the accumulation of γ-fatty acids in broiler meat. Six hundred one-day-old male chicks (Ross strain) from commercial broilers were divided randomly into 6 groups × 4 repeat pens. The broilers were fed experimental diets containing 4.0% tallow(control), 0.5% EPO, 0.7% mixed oil(EPO 70 : soy bean oil 30), 1.5% EPO, 3.0% EPO or 4.0% EPO for 35 days. There was a significant difference in body weight gain between the control and treatment groups except for the 0.5% EPO group (p<0.05). There was a significant difference in the percentage of thigh and breast weight against the carcass weight between control and treatment groups except for the 0.5% EPO group in the thigh and 0.5% EPO and 4.0% EPO groups in the breast weight (p<0.05). The saturated fatty acid levels of the skin and breast muscle lipid of the broilers fed diets containing EPO were significantly lower than that of the control group (p<0.05), while the level of unsaturated fatty acid was significantly higher than that of the control group (p<0.05). The γ-fatty acid(GLA, gamma-linolenic acid, 18:3n-6) level was particularly higher in the chicken meat lipids from the broilers fed EPO than in the control group (p< 0.05). This shows that feeding EPO to chicks can produce novel functional broiler meat that is enriched in gamma-linolenic acid.

The objective of this study was to determine the effect of dietary oils on the levels of the γ-linolenic acid in chicken meat lipids. Three hundred ten five, 1-d old, male, Ross strain, broiler chicks were fed for 35 d to compare diets containing evening primrose oil(EPO) and hemp seed oil(HO) to a control diet. Fatty acid composition of lipid from chicken skin, thigh and breast muscle were determined at the end of the trial. The level of γ-linolenic acid of lipids from chicken meat fed diets containing EPO or HO was significantly higher than that of the control group(p<0.05). The level of γ-linolenic acid of lipids from chicken skin was highest in the group, which had been fed the EPO 0.85%, followed in order by EPO 0.7%, 0.5%, EPO mixed oil, HO and HO mixed oil. There was a significant difference in the level of γ-linolenic acid of chicken skin between the control and treatment groups(p<0.05). The level of γ-linolenic acid of lipids from chicken thigh muscle was also similar to skin, and significantly higher than that of the control group(p<0.05). The level of γ-linolenic acid of lipids from chicken breast muscle was highest in the group, which had been fed the EPO 0.5%, followed in order by EPO 0.7%, 0.85%, HO 0.5% and HO mixed oil. There was a significant difference in the level of γ-linolenic acid of chicken breast muscle between the control and treatment groups(p<0.05).

The objective of this study was to determine the effect of dietary oils on the levels of the γ-linolenic acid in chicken meat lipids. Three hundred ten five, 1-d old, male, Ross strain, broiler chicks were fed for 35 d to compare diets containing evening primrose oil(EPO) and hemp seed oil(HO) to a control diet. Fatty acid composition of lipid from chicken skin, thigh and breast muscle were determined at the end of the trial. The level of γ-linolenic acid of lipids from chicken meat fed diets containing EPO or HO was significantly higher than that of the control group(p〈0.05). The level of γ-linolenic acid of lipids from chicken skin was highest in the group, which had been fed the EPO 0.85%, followed in order by EPO 0.7%, 0.5%, EPO mixed oil, HO and HO mixed oil. There was a significant difference in the level of γ-linolenic acid of chicken skin between the control and treatment groups(p〈0.05). The level of γ-linolenic acid of lipids from chicken thigh muscle was also similar to skin, and significantly higher than that of the control group(p〈0.05). The level of γ-linolenic acid of lipids from chicken breast muscle was highest in the group, which had been fed the EPO 0.5%, followed in order by EPO 0.7%, 0.85%, HO 0.5% and HO mixed oil. There was a significant difference in the level of γ-linolenic acid of chicken breast muscle between the control and treatment groups(p〈0.05).

The purpose of this study was to determine the effect of dietary γ-linolenic acid on plasma lipid metabolism and anti thrombotic activity in male Sprague Dwaley Strain rats. Rats weighing an average of 100~120g were fed a experimental diets containing 5% lard (saturated fatty acids), corn oil(linoleic acid), evening promise oil(EPO, 9% γ-linolenic acid) or borage oil(BO, 24% γ-linolenic acid) for 3Odays, respectively. Though there were no significant difference in the food intake among the groups, the body weight gain of the BO group was significantly lower than that of other group. The spleen weight of the lard group was significantly lower than that of other group. The bleeding time of the BO group was significantly longer than that of other group. The blood clotting time was significantly tended to long in EPO and BO groups compared with lard group. The plasma triacylglyceride and total cholesterol concentration were high in order of lard, com oil, EPO and BO, groups and there were significant differences among the groups. The plasma HDL-C concentrations were high in order of BO, EPO, com oil and lard groups and there were significant differences among the groups. The plasma LDL-C concentrations were significantly the highest in lard group, but the lowest in BO group. These data indicate that γ-linolenic acid has a antithrombotic activity, and decrease the plasma triacylglyceride, total cholesterol and LDL-C concentrations in rats.

100개의 토양시료로부터 120종의 곰팡이를 분리하여 이 중 GLA 생성능이 가장 우수한 균주를 선발하여 Fusarium 속으로 동정하였다. Fusarium 속이 생산하는 균체량과 지방질 함량은 각각 620㎎/100㎖, 63.5㎎/100㎖ 이었으며 전체 지방산 중 GLA가 차지하는 함량은 약 10.2% 이었다.

In order to investigate of the Influence of Mg2+, Ca2+ on α-linolenic acid converted into the eicosapentaenoic acid(EPA) and docosahexaenoic acid(DHA) forming in plasma lipid and in liver microsomes of rabbit, the animals were fed on the perila oil rich α-linolenic acid or sardine oil rich EPA and DBA diet for 4 weeks were examined. In plasma, liver lipid, Mg2+ was influenced on arachidonic acid(AA), EPA, DHA formative from α-linolenic acid in perilla oil, but stearic acid was increased, Ca2+ was Influenced on stearic acid increased and DHA was decreased. In phospholipid, Mg2+, Ca2+ was influenced on stearic acid increased and DHA was decreased in perilla oil.

To investigate the influence of saturated fats, α-linolenic acid, EPA and DHA on the lipid hydroperoxide concentration and fatty acid composition in liver microsomes and in plasma lipid of rabbits, the animals were fed on the perilla oil rich α-linolenic acid or sardine oil rich EPA and DHA diet for four weeks Were examined. The fatty acid composition of plasma lipid and liver microsomes of rabbits fed on the perilla oil diet was an accumulation of arachidonic acid(AA) 20:4 n-6, eicosapentaenoic acid(EPA) 20:5 n-3, and docosahexaenoic acid(DHA) 22:6 n-3, The fatty acid composition of plasma lipid and liver microsomes of rabbits fed on the sardine oil was an accumulation of α-linolenic acid(LNA) 18:3 n-3, and arachidonic acid(AA) 20:4. The p/s ratio of rabbits fed on the perilla oil diet changed from 7.4 to 2.27 for plasma lipid and 2.47 for liver microsomes. The concentration of lipid hydroperoxide was 3.48 nmol MDA/ml and 4.35 nmol MDA/ml for plasma lipid and liver microsomes, respectively, in perilla oil diet. The lipid hydroperoxide liver was 4.22 nmol MDA/ml and 67 nmol MDA/ml for plasma lipid and liver microsornes in sardine oil diet.

The aim of this study was to determine the effect of additional alpha-linolenic acid (ALA) supplementation during in vitro maturation (IVM) and culture (IVC) on nucleic maturation and embryo development of pigs. Cumulus-oocyte complexes (COCs) were incubated in IVM medium containing different concentration of ALA (25, 50 and 100 μM) for 44 h. After in vitro maturation, nuclear maturation of oocytes were evaluated by aceto-orcein stain. Mature oocytes with 50 μM ALA were fertilized and cultured in IVC medium with ALA (25, 50 and 100 μM) during early-embryogenesis (48 hours after fertilization). Then, embryos were cultured with 25 μM ALA during early embryogenesis and/or late embryogenesis (120 hours after early-embryogenesis). In results, oocyte maturation were significantly increased by 50 μM ALA treatment groups compared with control groups (p<0.05). Treatment of 25 μM ALA during early-embryogenesis enhanced cleavage rate of embryo compared with other groups (p<0.05), whereas formation and total cell number of blastocyst had no significant difference. Similarly, cleavage rate of embryos were increased by 25 μM ALA supplement during early- or late-embryogenesis than ALA treatment both stage of embryogenesis (p<0.05), but did not influence to blastocyst formation. Interestingly, total cell number of blastocyst were enhanced in ALA treatment group during early-embryogenesis. These findings indicated that ALA supplement enhance the nuclear maturation of oocyte and embryo development, however, excessive ALA could negatively influence. Therefore, we suggest that ALA is used for improvement of in vitro production of mammalian embryo and further study regarding with functional mechanism of ALA is needed.

Soybean has around 20% oil in total seed compound. Fatty acid concentration of soybean oil is about 12% palmitic acid, 4% stearic acid, 23% oleic acid (ω-9), 54% linoleic acid(ω-6) 54% and 8% linolenic acid(ω-3). To improve oxidative stability and quality of oil, the breeding programs mainly focused on reducing saturated fatty acids, increasing oleic acid and reducing linolenic acid in soybean oil. In plant oil, an essential fatty acid omega-3 fatty acid is in the form of α-linolenic acid (ALA) therefore, increasing ALA in soybean oil became one of the breeding goals for human health. In our research group, we have two breeding programs for concentration of ALA in soybean oil. Wild soybeans have almost twice ALA than that in cultivated soybeans. Introgression of alleles from wild soybean to cultivated soybean may lead to the increase of ALA in soybean seed oil for various applications. We developed several backcross populations by elite cultivars x wild soybean to select high ALA with good agronomic traits. In the case of low linolenic acid program, we developed an EMS (ethyl methane sulfonate) mutation population to select low ALA concentration line and found a mutant line with low ALA in seed oil. The scheme for developing high ALA concentration from wild soybean and molecular characterization for low ALA line will be discussed.

Scientific studies have shown that essential fatty acidintake can have a dramatic impact on human health. Soybean [Glycine max(L.) Merr.] oil from current commercial cultivars typically containsaround 8%linolenic acid (18:3) known as omega-3 fatty acid. Omega-3 fatty acid plays an important role to prevent cardiovascular disease and cancer. Relatively high 18:3 content in seed oil is a trait of the wild soybean (Glycine soja Sieb. and Zucc.) ancestor of modern soybean cultivars. Wild soybean is native to Korean peninsula and recently thousands of wild soybeans collected by soybean researchers in Korea. The objective of this study were to determine the linolenic acid content for wild soybean collection and to determine the stability of linolenic acid content derived from wild soybean over environments. Fatty acid profile for 1,806 wild soybean accessions collected from South Korea was determined by GC. The range of linolenic acid was 7.3 to 23.7% with an average 15.6%. We developed a recombinant inbred population from a cross PI483463 (wild soybean with 15% 18:3) and Hutcheson (cultivar with 8% 18:3). Three RILs, RIL156, RIL159 and RIL166, with high linolenic acid content (over 14%), parents and Williams 82 as checks were grown in nine environments over 2008-2011. Results showed that the content of linolenic acid for the PI483463, Hutcheson, and Williams 82 ranged from 14.8 to 17.1, 8.5 to 9.7, and 6.9 to 8.4 % and averaged 15.4, 9.2 and 8.0%, respectively. However selected RILs 156, 159, and 166 ranged from 10.7 to 15.7, 14 to 15.8, and 14.8 to 15.8, and averaged 13.9, 14.9, and 15.2, respectively. Among the tested accessions, RIL166 was the most stable with the lowest range and CV, and had a relatively lower stability coefficient value than other genotypes. Genes related to high linolenic acid from wild soybean may be useful in developing higher linolenic acid soybean genotypes and would broaden the use of soybean in food applications to improve human nutrition and health.