Most of the melon(Cucumis melo L.) breeding lines in Korea show andromonoecious (male-perfect flowers) sex expression, which requires laborious hand emasculation to produce the F1 seeds. There is a high demand for developing monoecious (male-female flowers) elite germplasm. The present study was carried out to develop molecular markers for selecting monoecious plants based on the CmACS-7 gene [a locus with 1-aminocyclopropane-1-carboxylic acid synthase(ACS) activity] responsible for ethylene synthesis and sex determination in melon. The full length sequences of the CmACS-7 were cloned from a monoecious inbred ‘Mo23’ and an andromonoecious inbred ‘Am24’. Sequence alignment revealed a major SNP(C170T) in exon1 and 18bp indel in intron4 of the CmACS-7, and a CAPS (SNP-C170T) and SCAR (ID4-18) were developed from the SNP and indel, respectively. A total of 453 F2 plants derived from ‘Mo23’ x ‘Am24’ were determined for their sex expression and genotyped using the SCAR marker. A Mendelian ratio of 3(monoecy): 1(andromonoecy) was observed from the F2 population, and sex type of 449 plants (except for four plants that showed incomplete monoecy) cosegregated with the SCAR marker, demonstrating that CmACS-7 is a single dominant gene conferring monoecy of ‘Mo23’. Allele variation of the CmACS-7 was evaluated by genotyping 114 melon accessions with diverse geographical origins using the CAPS and SCAR. C170T-SNP in exon1 of the CmACS-7 was highly conserved in melon germplasm and perfectly matched with the phenotype, whereas the 18bp-indel mutation in intron4 existed in various forms. The results demonstrated that CAPS marker SNP-C170T can be useful for marker-assisted selection(MAS) of monoecious melon plants

The whitebacked planthopper(WBPH), Sogatella furcifera is a serious pest of rice. The nymphs and adults suck phloem sap which causes reduced plant vigor, stunting, yellowing of leaves, delayed tillering in rice. This study was conducted to identify the optimum screening time for improved WBPH-associated QTL analysis and to develop the markers for use in breeding WBPH resistance. Resistance after 7 days infestation was observed in 100 lines(83.3%), after infestation for 14 days, resistance was observed in 14 lines(11.7%), and after infestation for 21 days, resistance was observed in 10 lines(8.3%). However, no after 14 days infestation was as similar as normal distribution in WBPH resistance. QTLs associated of the resistance detected in four regions on qWBPH1 and qWBPH8 in the intervals marker. After 7 days of infestation, the qWBPH1 was located in the interval RM3482-RM11966 and RM3709-RM11694 with LOD 4.0 and RM3709-RM11694 with LOD 3.5. After 14 days of infestation, The qWBPH1 was located in the interval RM3709-RM11694 with LOD 3.3. and RM3709-RM11694 with LOD 3.3. After 21 days of infestation, The qWBPH8 was located in the interval RM17699 with LOD 3.3. The QTLs on chromosome 1 was the most effective RM11694-RM11669 (LOD 3.3, variance 30%). The resistance lines were collected 10 plants of phenotye variation with genotype. The ratios of coincidence were used to determine resistance in 10 plants with phenotypic variation and a genotype of 8 markers. 3 markers were used: RM3482 on chr.1 represented 100%, RM8235 and RM11694 represented 80%, 90%, respectively, RM17699 on chr.8 represented 80% of the coincident ratio. These selected markers will be useful to rice breeding programs interested in new sources of WBPH resistance

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.

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

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.

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.

Wheat is one of the most important food crops in the world and provides 20% of nutritional requirement to the consumers in developing countries. The consumption per capita of wheat is persistent but the self-sufficiency has decreased owing to the less competitive world market and double cropping system with rice in Korea. This study was conducted to obtain baseline data on the selection lines with early maturity and good agronomic traits. Geumgangmil, a leading Korean variety, was crossed with CIMMYT’s line, and a DH (double haploid) line was released using pollens of Maize in Mexico. The 71 inbred lines were grown in fields at NICS, Miryang. The culm length of inbred lines were normally distributed from 37cm to 106cm, though that of the P1 (Geumgangmil) was 71 and P2 (CIMMYT’s Line) was 77cm. Especially, some lines were very short and measured below 50cm. The spike lengths of most of the lines were longer than that of P1 (7.5cm), but were normally distributed. The number of spikelet was less than that in P2 (19ea.), but was more than P1(10ea.). The heading day of most of the lines was between P1 (11th April) and P2 (25th April) and the maturity day of the lines was similar to P1 (1st June) but 5 lines showed 2~6 days shorter maturity period than that of P1. However, 20 lines were found to have 2~10 days longer maturity period than that of P2.

Fortification with vitamins in crops like rice is a continuing endeavor for geneticists and rice breeders. Tryptophan is one of the essential amino acids needed in human diet. In this study, we developed rice mutant lines using ethyl methane sulfonate (EMS) treatment in Korean cv. Donganbyeo and candidate rice lines were selected by insensitivity to the tryptophan analog, 5-methyltryptophan. One of the mutants has a 20-25 fold higher tryptophan level in mature seeds than wild type. To identify the mutations in anthranilate synthase genes, OASA1 and OASA2 sequences were generated. Moreover, mRNA expression levels of tryptophan biosynthesis related genes were examined. To further qualify the tryptophan fortification in rice, comparative assessment of cooking and eating quality was conducted with mutant lines and wild type. The moisture, viscosity, taste quality, protein content, amylose content and amino acid composition were similar with wild type. However, tryptophan contents in the mutant lines were higher than wild type as we targeted. The mutation present in AS gene of 5MT resistant rice may prove useful for the generation of crops with increased tryptophan contents and the mutation differences in AS sequences can be used for selection of mutant lines with high tryptophan level from large population.

Glucosinolates of Brassica rapa collection from Korea genebank were measured to determine total glucosinolate content and their variation of diverse glucosinolates; Around 100 accessions representing the different morphotypes and geographical origin of Brassica rapa were analysed. The principal component analysis was performed to evaluate the differences among morphotypes using the profiles of 14 glucosinolates identified from the leaves. DMRT test and box plots showed the significant difference between total glucosinolates of subspecies. Most of turnip accessions had higher gluconilates compared to the other type accessions, Chinese cabbage and pak choi. These accessions will be used for GWAS study for glucosinolate. Now they are being finger-printed by genotyping by sequencing (GBS). Among these accession, we selected a turnip accession with high amount of glucosinolate, K0466 and two Chinese cabbage accession with low amount of glucosinolate, K0015 and K0621. To analyse quantitative traits loci (QTL) for glucosinolate synthesis, these three accessions were fixed through microspore culture. Finally, six homozygous lines were selected and were crossed each other to make F1 hybrids. We just harvested F2 seeds and transferred doubled haploid plants to pots. QTL analysis for glucosinolate will be performed these F2 and DH population.

Internode elongation is an important agronomic trait in rice that is associated with lodging, yield, and flooding adaptation. We identified a novel rice mutant line showing shortened uppermost internode among the rice Ac/Ds insertional mutant population and named it shortened uppermost internode 4 (sui4). The phenotypes of F1 plants and F2 plants from the cross of sui4 with its original variety, Dongjin, indicated that the SUI4 gene shows incomplete dominance or semidominance. Because the Ds genotypes did not co-segregate with the sui4 phenotypes, we performed mapping of this gene with 273 F2 plants from a cross between sui4 and Milyang23. Primary mapping revealed that the SUI4 locus was located between the S07012 and S07015 markers on rice chromosome 7. Further fine mapping narrowed down the location of SUI4 to the 1.1-Mbp interval of RM1253-S07015. Using re-sequencing data of this mutant along with its original variety, Dongjin, and five other varieties, we found six sui4 specific SNPs occurred within the genic region of five genes in the fine-mapped interval. Among them, one SNP is in exon, while the other five SNPs are in intron. This SNP in exon occurred in the miR172 binding site of a gene encoding AP2 domain transcription factor, which seems to interrupt suppression of this gene activity by miR172. We isolated the genomic region of this gene from sui4 and transformed the wild type variety, Dongjin. The transgenic plants showed remarkably shortened internodes, which indicates that this AP2 domain transcription factor gene is the SUI4 gene.

An increasing preference for good eating quality of rice among consumers has become one of the important considerations in rice breeding. Amylose content of starch is one of the important factors of rice eating quality. Amylose composition is determined by the relative activity of soluble starch synthase (SSS) and granule-bound starch synthase (GBSS). This study focuses on modifying the expression of SSS1 gene which is responsible for amylopectin and amylose synthesis in rice by using RNA interference (RNAi) and antisense technology. The transgenic rice plants showed various amylose content (9-17%) in rice seed. Candidate rice lines were selected according to PCR, RNA expression and amylose contents analyses. A semi-quantitative RT-PCR was carried out to determine the expression level of SSS1 gene at several time points after the flowering of transgenic plants. Downregulation of SSS1 gene in transgenic rices was evident in the decreasing expression in rice grains over time. Accordingly, SEM micrographs analysis revealed uniform size with smooth curves starch granules in downregulation rice lines, in contrast with the non-uniform granules in wild type.

Onion(Allium cepa L.) is one of the most important crops to the fresh vegetable spices and the food processing industry in Korea. To evaluate genetic variation in onion, major agricultural characteristics and biochemical characteristics such as flowering day, harvest day, brix and so on. Phenolic compound compositions were analyzed for 44 accessions. Accessions in white color was about 92% which is the largest one followed by standard criteria. The sweetness of juice ranged from 6.1 to 19.3(in brix), and Brix range of 117 accessions was from 10.1 to 14.0. High performance liquid chromatography(HPLC) was used to identify the phenolic profile and quantify phenolic content in bulbs: quercetin, quercetin 7, 4’-diglucoside, quercetin 3-glucoside and quercetin 4’-glucoside were detected as major components. The total Quercetin and Quercetin glycoside content ranged between 123.59 and 1155.84 mg/100mg bulb freeze dried weight. The quercetin contents was up to 16.7mg/100g, and 7.41mg/100g in average. It is expected that the result of this study can be used for breeding more competitive species with respect to contents in functional chemicals.

Rice embryo contains valuable materials which are related to human health and industrial material, thus controlling embryo size is more and more important in the field of rice breeding. Especially, main health-aid components such as γ -aminobutyric acid (GABA), tocopherol and vitamins showed positive-correlation with embryo size. We obtained three enlarged embryo character mutants derived from Hwacheongbyeo (Korean japonica cultivar) by treatment of chemical mutagen, N-methyl-N-nitrosourea(MNU). These three mutants were named according to their embryo size as ge-m, ge, ge-s. The result of allelism test between Hwacheongbyeo, ge-m, ge and ge-s represented that the embryo size of ge and ge-s was controlled by the same gene(Giant embryo, GE). Through GE locus sequencing of three mutants, we found that each of ge and ge-s mutant has a point mutation, causing non synonymous amino acid substitution. On the other hand, ge-m mutant, the embryo of which featured intermediate size in between those of Hwacheongbyeo and ge, turned out to be non-allelic to the GE locus, suggesting it is likely a novel gene, which influences rice embryo development through a different mechanism than GE gene. Fine mapping of ge-m is currently in progress. This work was supported by a grant from the Next-Generation BioGreen 21 Program (Plant Molecular Breeding Center No. PJ008125), Rural Development Administration, Republic of Korea.

‘Muhan’ (Hordeum vulgare L.), a new ruminant-palatability forage barley cultivar, was developed by the breeding team at the Department of Rice and Winter Cereal Crop, National Institute of Crop Science, RDA in 2013. It was derived from the cross between SB921011-3B-16-2/IT0188556 and ‘Suwon 365’. Among the cross made in 2002, a promising line, SB02T2097-B-B-B-18, showed good characteristics in potential forage yield in the yield trial tested at Iksan in 2009 to 2010 designated as Iksan 465. The line in the Regional Yield Trials(RYT) tested in eight locations around Korea for three years from 2011 to 2013, and was released as the name of ‘Muhan’. It has the growth habit of group Ⅳ, erect plant type, green leaf and awnless spike. Its average heading and maturing dates were on May 4, and June 4, respectively, with similar to check cultivar ‘Youngyang’. The cultivar had 102cm of culm length, 711 spikes per m2 and it showed better rate of leaf, winter hardiness, and resistance to BaYMV than those of the check cultivar. The average forage yield of ‘Muhan’ was about 12.4 ton ha-1 in dry matter in paddy field. ‘Muhan’ also showed 6.6% of crude protein content, 28.6% of ADF (Acid Detergent Fiber), 50.1% of NDF (Neutral Detergent Fiber), and 66.3% of TDN (Total Digestible Nutrients), including higher grade of silage quality for whole crop barley

‘Jungmo2506’ (Hordeum vulgare L.), a new ruminant-palatability forage barley cultivar, was developed by the breeding team at the Department of Rice and Winter Cereal Crop, National Institute of Crop Science, RDA in 2013. It was derived from the cross between ‘Sunwoo’ and ‘Milyang 34’. Among the cross made in 2002, a promising line, SB02T2020-B-BB- 64, showed good characteristics in potential forage yield in the yield trial tested at Iksan in 2009 to 2010 designated as Iksan 463. The line in the Regional Yield Trials(RYT) tested in eight locations around Korea for three years from 2011 to 2013, and was released as the name of ‘Jungmo2506’. It has the growth habit of group Ⅳ, erect plant type, green leaf and short awn spike. Its average heading and maturing dates were on May 3, and June 2, respectively, with similar to check cultivar ‘Youngyang’. The cultivar had 99cm of culm length, 692 spikes per m2 and it showed better rate of leaf, winter hardiness, and resistance to BaYMV than those of the check cultivar. The average forage yield of ‘Jungmo2506’ was about 11.8 ton ha-1 in dry matter in paddy field. ‘Jungmo2506’ also showed 6.6% of crude protein content, 27.4% of ADF (Acid Detergent Fiber), 49.2% of NDF (Neutral Detergent Fiber), and 67.2% of TDN (Total Digestible Nutrients), including higher grade of silage quality for whole crop barley.

A new peanut(Arachis hypogaea ssp. fastigiata L.) variety “Hwaseon(Milyang50)” with the high yield potential was developed at the Department of Functional Crop, NICS, in Milyang in 2013. This was developed from the crossing line between cultivar “Iksan 31” with large grain and high quality and “SP9614” with short stem. “Hwaseon” which is Shinpung plant type has 41cm of main stem length and 10 branch number per plant. Each pod has two grains with long ellipse shape of brown testa and yield components is composed of 45 mature pods of per plant, 84g of 100-seed weight, 79% of pod shelling ratio in the regional yield trials(RYT). Seed quality showed 47.3% of crude oil and 27.6% of protein content. This variety also showed more resistant to early leaf spot, and stem rot, compared with check variety “Daekwang”. In the regional yield trials “Hwaseon” outyielded check variety by 12% with 4.78 MT/ha for kernel.

High oleate oil enhance flavors and shelf life and decrease coronary heart disease mortality. A new peanut(Arachis hypogaea ssp. fastigiata L.) variety “K-Ol(Milyang56)” with high oleic acid of fatty acid composition was developed at the Department of Functional Crop, NICS, in Milyang in 2013. This variety was developed from the crossing line between cultivar “Baekseon” with large grain and erect plant type and “F435-5” with high oleate and runner type. “K-Ol” which is Shinpung plant type has 43cm of main stem length and 10 branch number per plant. This also show more resistant to late leaf spot, web blotch and stem rot, compared with check variety “Daekwang”. Each pod has two grains with short ellipse shape of brown testa and yield components is composed of 46 mature pods of per plant, 74g of 100-seed weight, 76% of pod shelling ratio in the regional yield trials(RYT). For 3 year regional yield trials the average yield of “K-Ol” had 4.45 MT/ha for kernel similar to that of check variety. Seed quality show 50.7% of crude oil and 82.9% of oleic acid. The oil acidification of “K-Ol” measured by acid value(P-A value) variation for 3 hours at 180 ℃ showed one-ninth as low as ‘Daekwang’ do. This variety is expected to develop new demand area such as the improvement of cardiovascular disorders and shelf-life, and import substitution of olive oil.

A new peanut(Arachis hypogaea ssp. fastigiata L.) variety “Danwon(Milyang51)” with short stem length and lodging resistance was developed at the Department of Functional Crop, NICS, in Miryang in 2013. This variety was developed from the crossing line between cultivar “YG 75” with large grain and fewer branch number and “SP9617” with short stem length. “Danwon” which is Shinpung plant type has 32cm of main stem length and 13 branch number per plant. Each pod has two grains with long ellipse shape of brown testa and yield components is composed of 40 mature pods of per plant, 91g of 100-seed weight, 77% of pod shelling ratio in the regional yield trials(RYT). Seed quality show 48.7% of crude oil and 26.9% of protein content. “Danwon” has resistance to lodging in even double dense planting without yield reduction. This is more resistant to early and late leaf spot, web blotch and stem rot, compared with check variety “Daekwang”. In the regional yield trials “Danwon” outyielded check variety by 6% with 4.52 MT/ha for kernel.

The spikelet number per panicle (SPP) in rice is a quantitative trait controlled by numerous genes. To identify the alleles of these genes, we conducted sequencing of coding region or UTR of 13 genes which are known for controlling SPP trait, subjected to 48 rice cultivars covering 27 Indica, 17Japonica, and 4 Tongil types. The genes were EP3(ERECT PANICLE 3), RCN1(REDUCED CULM NUMBER 1), RCN2(REDUCED CULM NUMBER 2), FZP(FRIZZY PANICLE), LAX1(LAX PANICLE 1), MOC1(MONOCULM 1) APO1(ABERRANT PANICLE ORGANIZATION 1), APO2(ABERRANT PANICLE ORGANIZATION 2), DEP1(DENSE AND ERECT PANICLE 1), FON1(FLORAL ORGAN NUMBER 1), GHD8(GRAIN NUMBER, PLANT HEIGHT, AND HEADING DATE 8), GN1A(GRAIN NUMBER 1A), and HD1(HEADING DATE1). As a result, 1 synonymous SNP was found in EP3, RCN1 and RCN2. In LAX1 and MOC1, 1 non-synonymous SNP was identified. But none of SNP was found from the coding region of FZP. Moreover, 3, 6, 7, 8, 9 and 10 alleles were detected in APO1 and APO2, FON1, GHD8, GN1A, DEP1, and HD1 respectively. The multiple regression analysis revealed that GHD8, DEP1 and HD1 had strong effects on spikelet number of primary branch. In addition, HD1 had strong effects on spikelet number of secondary branch, and for total spikelet number per panicle as well. This work was supported by a grant from the Next-Generation BioGreen 21 Program (Plant Molecular Breeding Center No. PJ008125), Rural Development Administration, Republic of Korea.