Soybean [Glycine max. (L.) Merr] is one of the most important legumes in the world. However, soybean varieties are sensitive to flooding stress and their seed yields are substantially reduced in response to the flooding stress. 192 soybean germplasm collection was screened to identify flooding tolerant germplasm at an early vegetative growth stage (V1). Soybean plants at V1 stage were waterlogged for 4 to 10 days. To evaluate flooding tolerance, survival rate were investigated as a time dependent manner. Jangbaegkong, Danbaegkong, Sowonkong, Socheong2 and Suwon269 showed flooding tolerance, while Shillog, T201, T181, NTS1116 and HP-963 showed flooding sensitivity. We also investigated effects of flooding stress on soybean morphology. The adventitious root development was greatly increased in flooding tolerant plants compared to it in flooding sensitive plants. In addition, root length and root number were analyzed. The significant reduction of root length and root number was observed in flooding sensitive plants. Thus, these results indicate that the morphological changes in roots are important for acclimation to flooding stress. Taken together, the relationship between the morphological changes in the roots and flooding tolerance may be useful in selecting a flooding tolerant soybean germplasm.

Receptor mediated signal carriers play a critical role in regulation of plant defense and development. Rapid Alkalization Factor (RALF) is an important signaling family which has a role in plant growth and development. However, only few RALF polypeptides have been identified till date, mainly because of enormous efforts required for their isolation or identify their gene through mutational analysis. In this study, an extensive database search yield 39, 43, 34 and 23 potential RALF genes in Arabidopsis, rice, corn and soybeans, respectively. RALF genes are highly conserved across the plant species. A comprehensive analysis including the chromosomal location, gene structure, subcellular location, conserved motif, protein structure and promoter analysis was performed. RALF genes from four plants under study were divided in 7 groups based on phylogenetic analysis. In silico expression analysis of these genes, using microarray and EST data, reveled that these genes exhibit a variety of expression pattern. Furthermore, RALF genes showed distinct expression pattern under nitricoxide (NO) stress in Arabidopsis. This suggests a role of RALF genes in plant defense regulation. Our comprehensive analysis of RALF genes is a valuable resource that further elucidates the roles of RALF family members in plant growth and development. In addition, comparative genomics analyses deepen our understanding of the evolution of RALF gene family and will contribute to further genetics and genomics studies of other monocot and dicot plant species.

Preharvest sprouting resistance (PHS) causes the reduction of grain yield and also affects the quality of grains, resulting significant economic losses. PHS and its related traits were evaluated and observed in wide range among the 137 diverse rice accessions. To mine the associated signals for PHS resistance, genome wide association study (GWAS) was performed using phenotype data and whole genome resequencing data of 137 diverse rice accessions. This study not only could detect the previously identified dormancy and PHS associated genes but also explore the new candidate genes associated with the PHS and related traits. An example of them is seed dormancy 4 (Sdr4) gene which was found to be associated with germination % at day 14 (D14). This study provided the potential associated candidate genes which might be very useful to improve the PHS resistance in future rice breeding.

A sugary mutant with low total starch and high sugar content was compared with its wild type Sindongjin for grain-filling caryopses. In the present study, developing seeds of Sindongjin and sugary mutant from the 11th day after flowering (DAF) were subjected to RNA sequencing (RNA-Seq). A total of 30,385 and 32,243 genes were identified in Sindongjin and sugary mutant. Transcriptomic changes analysis showed that 7,713 differentially expressed genes (DEGs) (log2 Fold change ≥1, false discovery rate (FDR) ≤ 0.001) were identified based on our RNA-Seq data, with 7,239 genes up-regulated and 474 down-regulated in the sugary mutant. A large number of DEGs were found related to metabolic, biosynthesis of secondary metabolites, plant-pathogen interaction, plant hormone signal transduction and starch/sugar metabolism. Detailed pathway dissection and quantitative real time PCR (qRT-PCR) demonstrated that most genes involved in sucrose to starch synthesis are up-regulated, whereas the expression of the ADP-glucose pyrophosphorylase small subunit (OsAGPS2b) catalyzing the first committed step of starch biosynthesis was specifically inhibited during the grain-filling stage in sugary mutant. Further analysis suggested that the OsAGPS2b is a considerable candidate gene responsible for phenotype of sugary mutant.

Salt toxicity is the major factor limiting crop productivity in saline soils. Rice is an important staple food crop of nearly half of the world population and is well known to be a salt sensitive crop. The completion and enhanced annotations of rice genome sequence has provided the opportunity to study functional genomics of rice. With the rapid development of the biotechnology techniques, we can use more accurate and reliable methods to study the mechanism and function in different stress conditions. In present study, 295 rice accessions of diverse origin were re-sequenced and used for genome-wide association study (GWAS) with several germination-related traits, including germination percentage (GP), germination energy (GE), germination rate (GR), germination index (GI), salt tolerance index (STI) in salt tolerant germination stage. Phenotyping of the rice accessions were carried out at 200mM NaCl to screen salt tolerance levels. GWAS was applied to detect the associated genes related to salt tolerance in rice germination stage. Variations and haplotypes of the associated genes were detected and correlation between the phenotypes and genotypes were validated using qRT-PCR.

Bakanae disease is one of the most serious and oldest problems of rice production, which was first described in 1828 in Japan (Ito and Kimura 1931). This disease may infect rice plants from the pre-emergence stage to the mature stage, with severe infection of rice seeds resulting poor germination or withering (Iqbal et al. 2011). Under favorable environmental conditions, infected plants have the capacity to produce numerous conidia that subsequently infect proximate healthy plants, resulting in major yield loss (Ou 1985). One hundred sixty nine NILs, YR28297 (BC6F4) generated by five backcrosses of Shingwang with the genetic background of susceptible japonica variety, Ilpum were used for QTL analysis. Rice bakanae disease pathogen, CF283, was mainly used in this study and inoculation and evaluation of bakanae disease was performed with the method of the large-scale screening method developed by Kim et al. (2014). A major QTL for resistance against bakanae disease on chromosome 1 was identified using SSR marker, RM9, which explaining 65 % of the total phenotype variation. The major QTL designated as qBK1 and mapped to a 4.4 Mbp region between RM24 (19.30 Mb) and RM11295 (23.72 Mb). The results of this study are expected to provide useful information toward developing resistant rice lines to this detrimental fungal disease.

OsLPS is pollen specific gene that express at late stage of pollen development in rice. Based on microarray database, promoter region of two genes Os03g0106900 and Os03g0106500 were identified. The sequence of 2287bp and 2468bp upstream region of these genes were amplified and designated as OsLPS10 and OsLPS11. These promoters were fused with GUS-GFP reporter gene in a destination vector, pKGWFS7 and introduced into rice (Dongjin cultivar) and Arabidopsis (Col-0). The results of GUS assay showed different pattern of gene expression in pollen of rice and Arabidopsis. In Arabidopsis, the OsLPS10 gene strongly activated in young anther and not expressed in mature pollen. Pollen development analysis revealed GUS expression was detected at unicellular stage and strongest at the bicellular pollen developmental stage. No GUS signal was recorded in mature pollen. In case of OsLPS11, no GUS signal was detected in during pollen development of inflorescent. By contrast, in rice, the GUS expression pattern of OsLPS10 and OsLPS11 exhibited similar. GUS expression was first detectable in the anthers of spikelets at the bicellular stage and intensity increased in tricellular and mature pollen. The GUS signal was not detected in the anthers in unicellular microspores in both genes, OsLPS10 and OsLPS11. The results suggested that these genes were different activity in heterologous plant system, monocot and dicot. Complementation analysis and Cis-regulatory elements will be examined to illuminate the characteristic of these genes

Based on the results of microarray analysis we selected ten candidate genes that express in pollen at the early pollen developmental stage. By PCR amplification, the promoter region of these genes were amplified from rice genomic DNA (Nipponbare) and cloned into the destination pKGWFS7 vector via an entry vector, pDONR201. The characteristic of promoters were evaluated in Arabidopsis thaliana (Col-0) through GUS expression analysis. Fifty T2 plants respectively from each promoter were tested. Whole inflorescence of individual plant was stained with 1mM X-Gluc solution to observe tissue-specific GUS expression patterns. The results showed that all 10 promoters activated in pollen tissues. Among them six promoters expressed at the early developmental stage (unicellular) of pollen and the others expressed at both early (unicellular) and late pollen developmental stage (mature pollen). The results indicated that these promoters would be potential applicable for the studies of pollen function. Currently, we are performing these promoters analysis in rice transgenic plants as well as molecular characterization.

As one of the most important crop, rice is not only a staple food of half world’s population but a wonderful model plant, which has been leading the evolution and functional genomics study. The next-generation sequencing technology are expediting rice genomic study, by providing a simple but powerful way. In this study, we re-sequenced a core collection of 137 rice accessions from all over the world along with 158 Korean breeding varieties. Finally, 6.3G uniquely mapped reads were obtained, and about 10 million SNPs and ~1.2 million InDels were identified with average sequencing depth of 7.5X. These will help us to maximize our germplasm utilization and assists all the deep research in population dynamics and functional studies. Here, we’d like to show the approaches applied to resequencing data mining and on-going activities.

2-acetyl-1-pyrroline (2AP) was widely known as the principal aroma compound, it development in rice has been reported due to the loss of function of betaine aldehyde dehydrogenase gene (badh2) on chromosome 8. In previous study, a lot of haplotypes have been found of this gene, while only limited haplotypes have been proved as functional mutations. A total of 137 core set accessions, and additional 45 germplasms have been employed in this study. Finally, two new mutations have been found (3bp deletion in exon12 and C/A SNP in exon 10), and 23 haplotypes have been detected, most of them had strong relations with aroma formation. According to the sequence results, five functional markers have been developed, the markers showed a highly efficient in discriminating the special aromatic rice varieties, and displayed perfect co-segregation with the trait of fragrance in F2 population. Those new markers developed in the present study would be useful in molecular breeding of fragrant rice varieties. Based on the haplotypes, the further research is in progress.

In order to select a rice population with useful trait such as arsenic tolerance for crop improvement, we have developed 3000 M7 Targeting Induced Local Lesions IN Genomes (TILLING) lines by gamma ray (GR) irradiation treatment to a rice variety (cv. Donganbyeo). A total of 2 M7 lines exhibited the arsenic (AsV) tolerant phenotype (hereafter, named Arsenic Tolerant TILLING line 1 and 2, and designed as ATT1 and 2), in which the shoots and roots length of ATT lines were significantly longer than those of wild type (WT) during As(V) treatment. To survey the DNA polymorphism of these plants, we conducted the Whole genome resequencing with 10x coverage in ATT lines. By comparative analysis among ATT lines, we have identified the common DNA polymorphism such as 11,817 SNPs (49.83% in ATT1 and 48.35% in ATT2) and 30,618 InDels (86.72% in ATT1 and 86.23% in ATT2). Also, these mutants were showed the close relationships more than WT. To further study the changed amino acids of genes, we commonly identified the 758 genes for non-synonymous SNPs and 249 genes for changed codon InDels. These genes were mainly exhibited the enriched GO functions such as catalytic activity, nucleic acid binding and transferring phosphorus-containing groups. To determine the genes associated with arsenic-related mechanism in DNA polymorphism of ATT lines, we have retrieved the two structurally altered genes (Os11g47870 and Os03g19900) for metalloid As(V) detoxification toward induced genes in response to arsenic treatments by public microarray datasets. We suggest that As(V) tolerant phenotypes of ATT lines are certainly affected by structurally altered genes associated with phosphorus transferring and As(V) detoxification during GR treatment

Capsinoids, low-pungent compounds, have the same biological effects as capsaicinoids such as anticancer and anti-obesity. A precursor of capsinoids, vanillyl alcohol, is known to be produced by mutations in the p-aminotransferase (p-AMT) gene. In the previous study, SNU11-001 (C. chinense) containing high levels of capsinoids was found in germplasm collections of Seoul National University. We found that this collection has a unique mutation in the p-AMT gene. In order to develop a cultivar containing high capsinoids contents, marker-assisted foreground and background selection were performed in this study. Backcrossing is an effective breeding method for introducing useful traits to an elite cultivar. Compared to conventional backcrossing, marker-assisted backcrossing (MABC) is extremely useful for recovery of a recurrent parent’s genetic background. To obtain background selection markers, a total of 412 single nucleotide polymorphism (SNP) markers was screened to obtain polymorphic SNP markers between ‘Takanotsume (C. annuum)’ and ‘SNU11-001’. Of the 412 SNP markers, 96 polymorphic SNP markers evenly distributed in pepper genome were finally selected. Plants carrying the pAmt/pamt genotype were selected from a BC1F1 population using SCAR markers derived from the unique p-AMT mutation of SNU11-001. BC1F1 plants carrying the pAmt/pamt genotype were subjected to background selection. Multiple genotype analysis was done using Fluidigm platform (BioMark). Once we obtain plants carrying most similar genetic background to recurrent parent, capsinoids contents will be measured and another round of MABC be done to obtain plants containing high levels of capsinoids

Anaerobic germination (AG) is plays important role in submergence resistance which is an important trait for rice production in flood-prone lowland areas. Slow seed germination and delayed seedling establishment due to flooding become major problem for modern sowing methods such as direct seeding and environmental friendly good agricultural practices especially using young seedling age. In total, 137 diverse rice accessions were evaluated for anerobic germination ability. Submergence 1 (SUB1) which is induced ethylene response factors is suggestive because genes belonging to this gene family play a crucial role in rice tolerance to submergence. In this study, haplotype variations of three AG related genes, SUB1 (SUB1A, SUB1B, SUB1C) were examined using whole-genome resequencing data of 137 accessions of rice core set. The new SNPs and InDels found in the exon of the sub1 loci would be useful in developing markers to screen the varieties with strong anaerobic germination ability in the future molecular breeding

Chloroplast (cp) DNA sequence data are a versatile tool for plant identification, barcoding and establishing genetic relationships among plant species. Different chloroplast loci have been utilized to infer evolutionary relationship of plant species. Although the overall structure of the chloroplast genome is generally well conserved, a number of mutations have been observed. Thus, documentation of chloroplast sequence variation has also been an valuable asset in plant population and evolutionary studies for over two decades. Recently, advance in chloroplast genome assembly from whole genome NGS data has become available. In the present study, chloroplast variations among 295 diverse origin accessions were detected based on Oryza rufipogon, which thought to be the progenitor of cultivated rice.. Variation calling was carried out using the whole genome re-sequencing data of those accessions along with the five rice reference cp genomes. Phylogenetic and evolution analysis of the six references and 295 accessions were performed using the whole reference genome sequence and the variation data, respectively. Also, nucleotide polymorphisms of 295 rice accessions were validated by using previously characterized 50 ecotypes. Differential SNP frequency across the rice cp genome suggests a regional dependent preferential high variation occurrence during the evolution of chloroplast

Resveratrol rice Iksan526 was developed by overexpession of T-DNA (RB::P-Ubi::RS::T-NOS::P-35S::PAT::T-35S::LB) in rice variety Dongjin. To confirm one locus insertion of T-DNAs, Mendelian genetic analysis was carried out on selection marker bar gene and objective RS gene separately by using a F2 population derived from a cross of Dongjin/Iksan526 (T6). A total of 450 four-leaf-old plants from F2 population were treated by 0.3% basta, and a phenotypic separation ratio of 3:1 (321 survival: 129 dead, p>0.90) complied with Mendelian inheritance indicating one locus insertion of bar gene. Genotypic separation was analyzed by using PCR with specific primers for 300 plants, which were selected from 321 survival plants after phenotypic separation. Results revealed a ratio 1:2 of homologous to heterozygous (92:208, p>0.90), which further confirmed one locus insertion of RS gene. In addition, comparison on agronomic traits and resveratrol contents between transgenic rice and the donor variety were launched to evaluate the phenotypic performance over multi-generations (years).

Two carotenoid biosynthetic genes, phytoene synthase (Psy) and carotene desaturase (CrtI) linked via synthetic 2A sequence under control of CaMV 35S promoter (two T0 plants 5 and 6) or β- conglycinin promoter (three T0 plants 7, 13 and 16) were transformed into soybean variety Kwangan. After agronomic and phenotypic selection at early generations, T5 progeny of PAC soybean were analyzed by Southern blot to confirm T-DNA copy numbers. A total of 27 homologous lines derived from one of three T0 plants (line 7 under the control of β- conglycinin promoter) with one copy T-DNA insertion, were separated and planted into greenhouse. Flanking sequence analysis was carried out on one of homologous line 6-2-3 and results indicated the T-DNA was intergenic inserted into chromosome 14 from 10,873,131 to 10,872,998 base of soybean chromosome. T-DNA insertion structure, flanking sequence and inserted gene expressions need to be analyzed in the further study.

Rice is the major food for half of the world population. The nutrition component in rice is critical for improvement of people’s health. Vitamin E serves as important antioxidant by quenching the free radical intermediates and thus protects the cell membrane. Because of the high nutritional value and the benefits of vitamin E in human health, increasing the tocochromanol content of major agricultural crops has long been in the focus of breeding programs and genetic engineering approaches. The key genes involved in tocopherol biosynthesis have been elucidated in Arabidopsis and other model organisms. Quantitative trait locus (QTL) study performed in Arabidopsis suggested that some of these key genes and a few additional loci contribute to natural tocopherol variations. Identifying such genetic variations in rice, enrich our understanding of the genetic mechanisms controlling tocopherol variation, which can be directly applied to rice breeding programs. In this study, we used genome-wide association mapping with high-resolution density SNPs of rice core set to identify natural allelic variations, which contribute to tocopherol increase in rice

Mungbean (Vigna radiata) is a fast-growing, warm-season legume crop that is primarily cultivated in developing countries of Asia. We constructed a draft genome sequence of mungbean to facilitate genome research into the subgenus Ceratotropis and to enable a better understanding of the evolution of leguminous species. The draft genome sequence covers 80% of the estimated genome, of which 50.1% consists of repetitive sequences. In total, 22,427 high confidence protein-coding genes were predicted. Based on the de novo assembly of additional wild mungbean species, the divergence of what was eventually domesticated and the sampled wild mungbean species appears to have predated domestication. Moreover, the de novo assembly of a tetraploid Vigna species (Vigna reflexo-pilosa var. glabra) provided genomic evidence of a recent allopolyploid event. To further study speciation, we compared de novo RNA-seq assemblies of 22 accessions of 18 Vigna species and protein sets of Glycine max and Cajanus cajan. The species tree was constructed by a Bayesian Markov chain Monte Carlo method using highly confident orthologs shared by all 24 accessions. The present assembly of V. radiata var. radiata will facilitate genome research and accelerate molecular breeding of the subgenus Ceratotropis.

본 연구는 강낭콩(Phaseolus vulgaris, PV)의 종피색에 따른 항산화 활성 차이를 알아보고자 수행하였다. 유전자원 에 따른 16가지의 강낭콩을 적색 3종, 황색 1종, 적색얼룩무늬 3종, 흰색 얼룩무늬 1종, 회갈색얼룩무늬 2종, 흰색 2종, 회갈색 2종, 흑색2종으로 총 8가지 색으로 나누어 진행하였다. 연구결과를 요약하면 16가지 강낭콩을 콩 전체, 콩 종피 그리고 종피을 제외한 콩으로 분리하여 각각 80% 메탄올에 48시간 추출하였다. 추출 후 원액을 필터하여 실험에 사용하였다. 강낭콩에서 종피가 차지하는 비율은 6.93～10.20%이었으며, 총 폴리페놀 함량은 종피에서 7.05～180.33mg GAE/mL, 콩 전체에서 17.96～61.23mg GAE/mL, 그리고 콩 알맹이 16.58～22.24mg GAE/mL이었 다. 종피 추출물에서 PV-151(적색얼룩무늬)이 180.33mg/mL로 가장 높은 폴리페놀 함량을 보였고, PV-132(회갈 색)에서 164.67mg/mL, PV-78(회갈색얼룩무늬)에서 158.36mg/mL, PV-133(회갈색얼룩무늬)에서 158.19mg/mL, PV-126(적색)에서 147.93mg/mL, PV-343(흑색)에서 121.49mg/mL으로 나타났다. 흰색 강낭콩은 폴리페놀 함량이 미비하였다. DPPH 라디칼 소거능은 종피 추출물 2.5배 희석 하였을 때, PV-151(적색얼룩무늬), PV-343(흑색), PV-78(회갈색얼룩무늬), PV-132(회갈색)에서 90%이상 소거능을 보였으며 농도 의존적으로 활성이 감소하는 것 을 확인하였다. ABTS 라디칼 소거능은 종피 추출물을 10배 희석 하였을 때, PV-78(회갈색얼룩무늬)가 88.16%, PV-132(회갈색)가 81.29%, PV-151(적색얼룩무늬)가 95.26%, PV-343(흑색)이 80.11%의 소거능을 보였다. 이상의 결과로 보아 색이 다른 강낭콩 종피마다 가지고 있는 폴리페놀 함량이 다르며 각각 다른 항산화 활성을 나타냈다. 적색얼룩무늬, 회갈색얼룩무늬, 회갈색 그리고 흑색처럼 강낭콩 종피의 색이 진할수록 총 폴리페놀 함량과 더불 어 항산화 활성이 높은 것으로 나타났다.

국내에서 육성된 콩 91품종에 대하여 SSR마커를 이용하여 품종판별의 기초자료로 이용하고자 다형성이 높은 5개 의 Primer(Sat_043, Sat_036, Sat_022, Sat_088, Satt045)를 이용하여 5단계로 판별하였다. 5개마커의 총 대립인자수 는 64개이었고, 범위는 10-15개이었으며, 평균대립인자수는 12.8개이었다. 판별 1단계의 Sat_043으로 판별 하였을 때 육성품종 91품종 중에서 부석콩(172bp)의 1품종이 판별되었고, 판별 2단계의 Sat_036으로 판별하였을 때 호장 콩(90bp)등 34품종이, 3단계의 Sat_022로 단경콩(230bp)등 29품종이, 4단계의 Sat_088로신팔달콩(160bp)등 12품 종이, 5단계의 Satt_045로 새별콩(132bp) 등 6품종이 판별되었으며, 82품종이 판별되었다. 판별되지 않은 9품종은 형태적특성에 의하여 서로간에 판별되었다.