새로운 유전자원을 창출하기 위해 풍산나물콩에 EMS를 처리한 후 돌연변이 집단을 육성하고, 돌연변이 계통 중 지방산 함량에 대한 변이체를 찾아 콩 유전육종 재료를 선발하고자 실시한 결과를 요약하면 다음과 같다.M2 3,744개체중 형태적 변이를 보이는 1,000개체를 선발하여 M4 세대에서 난쟁이형 (3.3%), 엽형변이 (2.6%), 엽록소결핍 (1.5%), 꽃색변이 (1.1%), 엽형변이를 보이는 난쟁이형 (0.2%)의 변이가 관찰되었다. 야생형인 풍산나물콩의지방산 함량은 palmitic acid 11.6%, stearic acid 3.4%, oleicacid 25.3%, linoleic acid 52.0%, linolenic acid 8.1%를 나타내었다. M4 892 개체의 종자지방산 함량을 분석한 결과palmitic acid, stearic acid, oleic acid, linoleic acid, linolenicacid 함량은 각각 7.4~19.7%, 2.2~13.0%, 14.7~49.0%, 31.8~63.9%, 3.9~15.9%의 범위를 나타내었고, 10.8%, 3.8%25.3%, 52.0%, 8.1%의 평균을 보였다. 각각의 지방산 별로 선발된 돌연변이 개체들은 다음과 같다. 고 palmitic acid함량을 나타내는 PE1542 (17.1%), PE3058 (17.0%), 고 stearicacid 함량을 나타내는 PE977 (12.7%)와 저 stearic acid 함량을 나타내는 PE2166 (1.9%), 고 oleic acid 함량을 나타내는PE450 (44.4%), PE2742 (47.7%), PE3058 (33.4%), 저 linolenicacid 함량을 나타내는 PE594 (4.6%), PE1690 (3.7%)와 고linolenic acid 함량을 나타내는 PE2166 (12.6%) 등이 선발되었다.

한국자원 128개와 중국자원 214개 및 미국자원 37개를 포함한 총 379개의 국내외 콩 유전자원에 대한 지방 함량과 지방산 조성을 평가하고 유용자원을 선발하여 고품질 콩 품종 개발의 기초자료로 활용코자 수행되었다. 한국자원과 중국자원은 농촌진흥청 농업유전자원센터로부터, 미국자원은 경북대학교 식물유전육종연구실로부터 분양 받았다. 콩 유전자원의 지방함량은 평균 19.1%이었고, 12.7∼26.0%의 범위이었고, 포화지방산인 palmitic acid의 함량은 평균 10.6%이었고 범위는 4.1∼18.1%, stearic acid의 함량은 평균 2.9%이었고 범위는 1.9∼12.7%이었다. 불포화지방산인 oleic acid의 함량은 평균 28.2%이었고 범위는 15.3∼56%, linoleic acid의 함량은 평균 51.2%이었고 범위는 24.4∼68%, linolenic acid의 함량은 평균 7.1%이었고 범위는 3.4∼11.1%이었다. linoleic (ω-6)/linolenic(ω-3) acid 함량의 비율은 평균 7.5이었고 범위는 3.8∼17이었다. 포화지방산이 평균 13.5%이었고 범위는 6.7∼21.7%, 그리고 불포화지방산의 함량은 평균 86.5%이었고, 범위는 78.3∼93.4%이었다. 고지방품종 개발을 위한 지방함량이 25%이상인 고지방함량 자원으로는 중국자원인 Suinong 14, Dongnong 434 및 Dongnong 46의 3개이었다. Oleic acid 함량이 50%이상인 고 oleic acid함량 자원으로는 미국자원인 KLG12073과 KLG12074, 중국자원인 Jilin 14 및 한국자원인 Kwangan과 Bongeui의 5개이었고, linoleic acid의 함량이 60%이상인 고 linoleic acid 함량 자원으로는 미국자원 KLG12083과 KLG12081을 포함한 8개이었다. Linolenic acid의 함량이 4%이하인 저 linolenic acid 함량 자원으로는 미국자원 KLG12093와 KLG12096을 포함한 7개이었다. Linoleic acid(ω-6)/linolenic acid(ω-3)의 비율이 4.5이하로 낮은 자원은 미국자원 KLG12074 (3.8)와 KLG12073 (3.8), 한국자원 Kwangan (3.8)와 Bongeui (4.5) 및 중국자원 Dongnong 47 (4.3)의 5개이었다.

The wild relatives of soybean [Glycine soja Sieb. and Zucc.] have curly/wavy nature whereas cultivated varieties are upright. Such morphological characteristics have agronomic importance too. To investigate the molecular mechanism of development contributing to coiled morphology, screening was carried out to look for Arabidopsis mutants in activation tagging lines obtained by activation T-DNA treatment that have curly/wavy morphology. A mutant named Coiled Branch 1 (cbr1), is found to have a wavy and curly morphology with coiling branches. Plasmid rescue and genomic southern blot analysis revealed the site of T-DNA insertion in the genome. RT-PCR was performed to monitor expression levels of the genes adjacent to the T-DNA integration sites, and showed the activation of an E3 ubiquitin ligase gene. Database search showed that the gene with the RING domain belongs to a family of E3 ubiquitin ligases. Complementation test by overexpression and RNA interference of the gene was also carried out. The complementation test results showed that the novel gene activation tagging affected the cbr1 mutant phenotypes. Ubiquitylation has been linked virtually to every cellular process including plant development. E3 ubiquitin ligase has been reported to recognize target proteins that are to be ubiquinated for further degradation by the proteasome complex. Further, more detailed studies are needed to identify the specific substrate(s) of the novel E3 ubiquitin ligase gene.

Soybean [Glycine max (L.) Merr.] seeds are abundant in high-quality proteins and fats. In addition, soybean seeds are also rich in secondary metabolites, such as isoflavones, lecithin, and saponins. Triterpene saponins are major components of these physiologically active metabolites in soybean seeds. Soybean saponins are classified as group A and DDMP saponins. Among them group A saponins are undesirable component of food products due to bitterness and astringency and also cause foaming in tofu production. Whereas, DDMP saponins and their derivatives are less bitter and astringent and beneficial to human health when consumed as regular diet. Therefore, reducing the group A saponins or increasing the DDMP saponins are required to improve the food quality. The present study focused to identify and characterize the gene which is encoding a protein responsible for biosynthesis of DDMP saponins. EMS mutant lines (sg-7-1 & sg-7-2) which lack DDMP saponins were developed. The breeding cross has been made with these two mutants with two cultivars, Pungsannamul and Wooram to study the segregation and genetic linkage analysis, respectively. The segregation analysis showed that the mutant phenotype is controlled by single recessive gene. TLC analysis for phenotyping F2 population of Wooram X sg-7-1 showed mutant, wild and heterozygous types. To surprise two more patterns were detected and they were named as strange type1 (ST1) and strange type2 (ST2). Further, SSR marker analysis will be carried out to locate the gene which encoding a protein responsible for biosynthesis of DDMP saponins.

Soybean germplasm have diverse accessions with great variation in their ability to survive and reproduce under salt stress conditions. In general, cultivated soybeans are more sensitive to salt stress than their wild relatives, however exceptions are found in both the groups. These variations in response to salt stress makes soybean germplasm an interesting collection of genetic resources to be explored for the identification of salt-tolerance genes, and their mechanism of action. Here, in this report we presented a data showing differential response of selected accessions of both cultivated and wild soybeans to salt stress. Two modes of salt treatment; gradual salt stress (GS) as well as salt shock (SS) were used in this study. The GS was found more effective in finding the difference in response of soybean accessions to salt stress. Various genetic marker based methods are in use to identify and isolate the potential genes contributing to the salt tolerance in soybean. Even then there is a paucity of knowledge on the key genes contributing to the salt tolerance in soybean. We expect that a recently developed functional screen based method, like yeast based functional screen, using cDNA library generated from different salt tolerant accessions of soybean could lead to identification of novel genes responsible for salt tolerance in soybean. Also, we propose for the use of RNA isolated from different stages of GS and SS for making cDNA library to be used for functional screening.

Soybean [Glycine max (L.) Merr.] have a variety of flower colors which are controlled by six different genes (W1,W2,W3,W4,Wm, and Wp). Among these genes, mutation in W3 gene causes near white flowers in the background of w4 genotype whereas the genotype W3w4 does purple throat flowers. Earlier studies showed that dihydroflavonol 4-reductase1 (DFR1) gene was closely linked to the flower color variants for W3 locus. In order to find out the W3 gene responsible for w3 phenotype, we first, studied the candidate gene Glyma14g07940 (DFR1) which is having 100% similarity with DFR probe sequence. Sequence analysis of DFR1 between W3 and w3 soybeans showed one base substitution in exon 6 of w3 mutant soybean resulting in one amino acid change in the amino acid sequence. However, comparison of amino acid sequences of DFR proteins from various crop plants showed that there is no functional change in the protein. Besides, the promoter analysis showed that, 311 bp of indel was traced in 5’-upstream promoter region of DFR1 gene in the w3 mutant. Here, we show that the near white or purple throat phenotypes in G. max is associated with existence or nonexistence of indel at 5’- upstream promoter region and low or high expression of DFR1, respectively. These results suggest that w3 phenotype may be caused by certain regulator of DFR1 gene located near or distant from DFR1 in G. max. In further study, we need to check the correlation between promoter indel with W3 expression level through GUS analysis.

Soybean [Glycine max (L.) Merr.] is a major agricultural crop widely used for providing human and animal food owing to its high protein and oil content. For this reason, they have been consumed in Asia and world greatly and demand is ever increasing. Soybean is classified as a moderately salt-sensitive crop and its production is greatly affected due to increasing salinity stress. About 8 % of the world’s total land is salt-affected. In Korea, around 9 % of total agricultural land (approximately 130,000ha) was reclaimed since 1960's. In order to meet the demand for soybean and to solve arable land shortage problem, it is unavoidable to cultivate soybean in salt-affected soils. Fortunately, soybean germplasm has been shown to have salt-tolerant phenotypes, which have been used to identify the salt-tolerant genes. GmCHX1, a novel ion transporter, is one of the genes known to confer salt tolerance in soybeans. Present study was conducted to understand the effects of sequence variations of GmCHX1, on salt tolerance in wild and cultivated soybeans. A total of 1026 (301 lines of G. max and 725 lines of G. soja) lines were phenotyped for salt tolerance in greenhouse conditions. At the V1-V2 growth stage, the plants were treated with 100mM NaCl solution for two weeks and thereafter the response was measured depending on leaf scorch score (1-health, 3-mid, 5-dead). About 20 lines found to show tolerance to saline conditions and were selected for sequence analysis of GmCHX1. Most of the haplotypes detected in this study corresponded with the haplotype patterns in previous studies. However, several lines showed different patterns of polymorphism in the coding region, suggesting that sequencing of more lines and analysis for the polymorphism in GmCHX1 is needed in order to identify new haplotypes that could confer greater salt tolerance.

Foxglove aphid, Aulacorthum solani (Kaltenbach), is a Hemipteran insect that infected a wide variety of plants worldwide and caused serious yield losses in crops. The objective of this study was to identify the putative QTL for foxglove aphid resistance in wild soybean, PI 366121, (Glycine soja Sieb. and Zucc.). One hundred and forty one F2-derived F8 recombinant inbred lines developed from a cross of susceptible Williams 82 and resistant PI 366121, were used. The phenotyping of antibiosis and antixenosis was done through choice and no-choice assays with total plant damage (TPD) and primary infestation leaf damage (PLD); a genome-wide molecular linkage map was constructed with 504 single nucleotide polymorphism markers utilizing a GoldenGate assay. Using inclusive composite interval mapping analysis for foxglove aphid resistance, one major candidate QTL on chromosome 7 and 3 minor QTL regions on chromosome 3, 6 and 18 were identified. The major QTL on chromosome 7 showed both antixenosis and antibiosis resistance responses. However, the minor QTLs showed only antixenosis resistance response. The major QTL mapped to a different chromosome than the previously identified foxglove aphid resistance QTL, Raso1, from the cultivar Adams. Also, the responses to the Korea biotype foxglove aphid were different for Raso1, and the gene from PI 366121 against the Korea biotype foxglove aphid were different. Thus the foxglove aphid resistance gene from PI 366121 was determined to be an independent gene to Raso1 and designated to Raso2. This result could be useful in breeding for new foxglove aphid resistant soybean cultivars.

Shoot-fresh-weight (SFW) is one of the parameters, used to estimate the total plant biomass yield in soybean. Understanding the genetic and molecular basis of SFW could help increase the total biomass production. In this particular study, we identified QTLs associated with SFW in a Recombinant Inbred Line (RIL) population derived from interspecific cross of PI483463 and Hutcheson. A total of 551 (535 SNP and 16 SSR) markers, were found to be polymorphic between the parental lines and were used to screen the RILs to develop the genetic map. Linkage analysis and QTL mapping were performed using with the software QTL IciMapping version 4.0, with the minimum LOD score of 3.0 and estimating the likelihood of a QTL and its corresponding effects at every 1cM. QTLs with LOD value > threshold LOD, as determined by 1000 permutation tests at p > 0.05 were considered as significant QTLs. The analysis identified a total of 5 QTLs associated with shoot fresh weight over two environments, with the phenotypic variation (PV) ranging from 6.34 to 21.32%, and the additive effect from -0.54 to 0.33. Among these QTLs, qFW1314_19_1 had the largest LOD scores, with PV of 21.32%. Interestingly, three QTLs, qFW2013_19_1, qFW2014_19_1, and qFW1314_19_1 identified on chromosome 19(L), showed negative additive effects, indicating the contribution from the wild parent PI483463. The QTLs identified in this study can be the targets to identify the candidate genes for the SFW and can help in developing cultivars with increased biomass potential.

Foxglove aphid, Aulacorthum solani (Kaltenbach), is a Hemipteran insect that infected a wide variety of plants worldwide and caused serious yield losses in crops. The foxglove aphid resistance gene, Raso2 was previously mapped from PI 366121 (Glycine soja Sieb. and Zucc.) to a 26cM marker interval on soybean chromosome 7. The development of additional genetic markers, which are mapped closer to Raso2 were required to accurately position the gene to improve the effectiveness of marker assisted selection. The objective of this study was to narrow down the putative QTL region, which is responsible to foxglove aphid resistance in PI366121 using recently developed high-density 180K Axiom SoyaSNP genotyping array. One hundred and forty one F8-derived F12 recombinant inbred lines developed from a cross of susceptible Williams 82 and resistant PI 366121, were used to generate a fine map of Raso2 interval. The phenotyping of antibiosis and antixenosis was done through choice and no-choice assays with total plant damage (TPD) and primary infestation leaf damage (PLD). The composite interval mapping analysis showed that the physical interval between two flanking makers, which was corresponding to Raso2, was narrowed down to 500kb on the Williams 82 genome assembly (Glyma2.0), instead of 4Mb in the previous report using Goldengate assay. In the Raso2 interval, there are about 60 candidate genes, including 4 of NBS-containing putative R genes. This result could be useful in breeding for new foxglove aphid resistant soybean cultivars.

Seed weight (SW), often expressed as 100-seed weight (HSW), is an important yield component in soybean and has been found to show positive correlation with seed yield. It is shown to behave as a quantitative trait controlled by many loci that are largely unclear. In this study, we represent the identification of chromosomal regions controlling the seed weight in soybean. We used a Recombinant Inbred Line (RIL) population, consisting of 188 lines derived from a cross of a wild soybean PI483463 (HSW: 0.85g) and a cultivated soybean cultivar Hutcheson (HSW: 14.05g) to identify the chromosomal regions controlling the SW trait. The population, along with parental samples and check, William82 (HSW: 21.2g) was grown for four years and phenotype data was recorded postharvest. A total of 535 SNP and 16 SSR markers, polymorphic between the parents were employed to genotype the RILs using Golden gate assay to develop the linkage map. Whole genome QTL scanning identified a total of 17 QTLs, spanning 10 chromosomes for the 100-seed weight. All these QTLs explained phenotypic variation (PV) in the range of 3.77 to 12.33%. Of the 17 QTLs, 2 QTLs qSWA1-1 and qSWD2-1, found to be the consistent QTLs, expressing in all the four environments. The QTL qSWD2-1 explained highest contribution to the total PV with 10.04 -12.23 %. The remaining 15 QTLs were identified in at least one environment with PV ranging up to 10.39%. The findings from this study will provide useful information to understand the genetic and molecular basis of SW and facilitate further genomic research leading to the yield improvements in soybean.

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.

A total area of reclaimed land in Korea is about 135,100 ha, which occupies 9 % of total arable land. Soybean is one of the most important crop in Korea and demand for the crop is increasing, while the country’s self-sufficiency is very low, around 9 %. If it’s possible to cultivate soybean in reclaimed land, it would increase self-sufficiency of the soybean. However, there are difficulties to cultivate soybean in reclaimed land because of excessive level of salinity in the soil, to prevent this barrier in saline soils, it is necessary to develop salt tolerant soybean cultivar. This research was conducted to select salt tolerant lines derived from PI 483463 (salt tolerant wild soybean accession). The F1 (Hutcheson × PI 483463) was backcrossed with Hutcheson and Wooram (salt sensitive soybean cultivar). For marker assisted selection and salt reaction phenotyping, randomly selected BC1F1 seeds from two backcross populations were planted in 11 cm tall tray. At the V1 growth stage, DNA was sampled with FTA card. The genomic DNA and SSR marker, BARCSOYSSR_03_1348, were used for PCR amplification and the result was checked through electrophoresis. The trays with BC1F1 plants were immersed in 100 mM NaCl solution up to the bottom third of the trays directly after the DNA extraction. After two weeks, phenotype was measured depending on leaf scorch degree. Through this research, 25 dominant homozygote lines and 22 heterozygote lines from Hutcheson backcross population and 28 dominant homozygote lines and 37 heterozygote lines from Wooram backcross population were selected. These lines will be used for developing soybean with salt tolerance

Soybeans have been the favored livestock forage for centuries. However, little studies have been succeed in estimating forage quality of soybean by near-infrared reflectance spectroscopy (NIRS). To establish NIR equations for soybean forage quality, 353 forage soybean samples, including an 181 recombinant inbred line population derived from PI 483463 (G. soja) ´ Hutcheson (G. max), 104 cultivated soybeans (G. max) and 68 wild soybeans (G. soja) were used to develop NIR for four quality parameters: crude protein (CP), crude fat (CF), neutral detergent fiber (NDF), and acid detergent fiber (ADF). Two NIR spectroscopy equations developed for CP and CF (2,5,5,1; multiple scatter correction [MSC]) and for NDF and ADF (1,4,4,1; MSC) were the best prediction equations for estimating these parameters. The coefficients of determination in external validation set (r2) were 0.934 for CF, 0.909 for CP, 0.767 for NDF, and 0.748 for ADF. The relative predictive determinant (RPD) ratios for MSC (2,5,5,1) calibration indicate that the CP (3.34) and CF (3.45) equations were acceptable for quantitative prediction of soybean forage quality, whereas the NDF (2.34) and ADF (1.97) equations were useful for screening purposes. The NIR calibration equations developed in this study will be useful in predicting the contents of forage qualities and in breeding soybean for forage

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.

Soybean is desirable as a forage crop because of it has high protein and oil concentration. Wild soybean, a progenitor of cultivated soybean, has a softer stem and higher protein content in seed than cultivated soybean. There is little information on yield and forage quality for wild soybean and its derivatives. The objective of this study was to determine the forage yield and quality of wild soybeans and selected soybeans derived from a cross G. max ×G. soja. Forage yield and quality were assessed for three grain soybean cultivars, three wild soybeans and three selected lines from G. max×G. soja. Forage quality attributes such as crude protein (CP), crude fat (CF), neutral detergent fiber (NDF), acid detergent fiber (ADF), digestible dry matter (DDM), dry matter intake (DMI) and relative feed value (RFV) were determined at the R2, R4 and R6 developmental stages. Forage yield and CF were highest at stage R6 in G. max, G. soja and selected G. max×G. soja lines. CP content was similar between R2 and R4 but increased sharply after R4 and peaked at R6 in G. max and selected lines from G. soja×G. max. On the other hand, CP content was similar between R4 and R6 stage in wild soybeans. Generally, NDF and ADF were highest at stage R4 but decreased at stage R6. DDM, DMI, and RFV increased between R4 and R6. These results suggest that R6 was the optimal harvest stage to provide forage of highest quality and yield. A study was conducted in 2011 to evaluate forage yield and quality at stage R6 in 25 lines from PI483463 (G. soja)×Hutcheson (G. max) and four cultivated grain soybeans. Hutcheson had the highest forage yield with 24.7t/ha infresh weight (FW) among grain soybeans. Line W11 had the highest forage yield(25.7t/ha,FW) among G. soja×G. max selections and four other lines had similar forage yield compared to Hutcheson. Generally the 25 lines from this G. max×G. soja cross had thinner main stems and branches than cultivated soybeans. When the 25 lines were evaluated for their feed quality as per forage grade by AFGC, nine lines rated prime grade and all 25 lines were classified as forage Grade 1. Results of this study indicate crosses between wild and cultivated soybean show promise for improving soybean as a forage crop.