Mutagenesis approach in combination with whole genome sequencing has become an import role in genetic and molecular biological study and breeding of crop plants. In this study, we screened the fast neutron M4 10,000 soybean mutant plants based on morphological phenotypes of agronomically important traits and characterized the mutant of interest using resequencing. Fast neutron radiation has been known to be a very effective mutagen to cause large deletion in genome. The screened mutant showed abnormal phenotypes in plant heights, seed sizes, color of leaves, number of leaves, maturity and number of branches etc. Among them, the mutant displaying short plant height and bush type of growth habit was selected for identification of the altered genomic regions. Analysis of deletion sites of genome in interesting soybean mutant was performed using next generation sequencer Illumina Hi-seq. Mutant sequence reads generated by paired-end shotgun library were mapped on a draft soybean reference soybean (G. max cv. Williams 82). The paired-end DNA sequences of 21.6 Gb produced by Illumina Hi-seq produced 21 fold sequence depth. Among the predicted deletion sites, total 3 deletion regions confirmed by PCR. Glyma03g02390 gene and Glyma03g03560 gene were involved in the deletion regions. Glyma03g02390 gene was related to AMP binding, catalytic activity, cofactor binding and metabolic process of cell growth and Glyma03g03560 gene was concerned to oxygen binding, defense response to bacterium, and especially process of indole acetic acid (IAA) biosynthesis. These genes detected in this mutant will be studied about their molecular function in stunted phenotype.

NABIC(National Agricultural Biotechnology Information Center) established integrated management system of agricultural omics information to achieve the agricultural bio-information resources of Korea. The amount of bio-information is enormously increasing due to emergence of NGS(Next Generation Sequencing) technology. We has building, maintaining and providing agricultural bio-information databases and information services. Various data type for submission is available such as genome, proteome, transcriptome, metabolome, molecular marker, etc. We issue the submission confirmation which is available for research achievement. Currently, the amount of data submitted on our system is 4.3Tb. We are planning to integrate genome annotation system and NGS analysis system this year. The Agricultural Omics Information Submission System is available through web site(http://nabic.naas.go.kr/submission).

Although a great deal of rice proteomic research has been conducted, there are relatively few studies specifically addressing the rice grain proteome. The existing rice grain proteomic research has focused on the identification of deferentially expressed proteins. Here, we performed comparative shotgun proteomic analysis of rice grain development to construct an in-depth proteome reference map, to reveal the expression patterns of the identified proteins, and to detect proteins that are expressed deferentially during grain development. A Korean rice variety, Ilpumbyeo was used. Proteins were extracted from rice grains 10, 20, and 30 days after flowering, as well as from mature grains. The protein expression patterns were revealed by a quantitative shotgun proteoemic analysis. By merging all of the identified proteins in this study, we identified 4,172 non-redundant. A Genome Ontology category enrichment analysis for the 4,172 proteins revealed that 52 categories were enriched, including the carbohydrate metabolic process, transport, localization, lipid metabolic process, and secondary metabolic process. The relative abundances of the 1,784 reproducibly identified proteins were compared to detect 484 differentially expressed proteins during rice grain development. Clustering analysis and Genome Ontology category enrichment analysis revealed that proteins involved in the metabolic process were enriched through all stages of development, suggesting that proteome changes occurred even in the desiccation phase. Interestingly, enrichments of proteins involved in protein folding were detected in the desiccation phase and in fully mature grain.

The amount of genetic variability of a species is essential for its survival and adaptation in different environments, and studies of genetic diversity using molecular markers are necessary to understand the genetic structure of a population and to orientate effective strategies of germplasm conservation. The aim of current study was to determine the SSR markers that can be used rapidly and reliably to evaluated the pepper of Bulgaria landraces, and applied the markers to assement of introduce genetic diversity of the pepper germplasm. We used 22 polymorphic microsatellite markers to analysis of genetic diversity within 61 pepper collection of Bulgaria landraces germplasm, all SSR primers pairs produced 80 polymorphic and reproducible amplification fragments. An average value of polymorphic information contents (PIC) were 0.334 with a range of 0.061 to 0.63. The mean values of observed (HO) and expected heterozygosity (HE) were 0.383 and 0.154, respectively, indicating a considerable amount of polymorphism within this collection. A genetic distance-based phylogeny grouped into three distinct groups, which was the landrace, moderate and wilde type, genetic distance (GD) value was 0.540. An average day of flowering time was 53 days with a range of 45 to 60 days. The everage od fruit length and width were 9.38cm with a range 2.1 to 23.6cm, and 3.51cm with a range 0.6 to 8.9cm, respectively. Molecular data were complemented with morphological measurements according to the descriptor list for the pepper collection of Bulgaria landraces germplasm.

The application of next generation sequencing technologies allows us to discover the high levels of DNA polymorphism throughout a genome, e.g., single nucleotide polymorphisms (SNPs), and insertions and deletions (InDels). We performed whole-genome resequencing of a Korean rice cultivar (cv. Donganbyeo) and then obtained the sequences of covered 366,042,872 bp (96.63%) with average mapped read depth of 34.17 on 382,788,128 bp of the Japanese cultivar genome (cv. Nipponbare). We characterized the polymorphisms of 173,711 SNPs, 295,334 insertions and 40,642 deletions based on the comparison of both genomes. About 11.5% and 17.8% of the annotated total SNPs were presented in the regions of 1kb upstreams and genes, respectively. The annotated InDels in gene regions were similar with 15.5% insertion (4,588) and 15.9% (5,100) deletions, but not in 1kb upstream regions with 9.0% insertion (2,662) and 14.3% deletions (5,100). In addition, the Korea rice genome sequences were mapped on individual chromosome, resulted that SNPs were shown with different frequencies from each chromosome. The InDels distributions on individual chromosomes exhibited similar pattern as compared to those of SNPs. Some gene families such as NB-ARC (NB-LRR), F-box, RLK (serine/threonine protein kinase) and Zinc-finger (RING) for SNPs occurred the similar pattern with those of Arabidopsis. These results might be useful for better understanding the genome structure and genetic diversity of the Korean rice cultivars.

We have identified ATTIRTA1 transposon, a kind of mariner-type DNA transposon from Brassica rapa genome. A total of 811 inverted-terminal repeat, ITR consisting of the both terminal on ATTIRTA1 transposon were found from B. rapa v1.1 sequence. Among them 616 ITR were paired by two in each transposon, indicating three quarters of the transposon exists in original form. Around 10 percentage of the transposon, 82 ITR was located in gene, expecially only in intron. Using these ATTRRTA1, we developed a display system modified from AFLP technique and applied for this system to analyze genetic diversity of Korea Brassica rapa core collection. The collection includes 220 accessions representing the different morphotypes and geographical origin. The analysis of population structure revealed five subgroups and the clustering patterns matched well with their morphological traits. ATTIRTA1 transposon display seems useful marker system for studying genetic relationships. Presently we have profiled the components and contents of glucosinolate in the core collection to analyze genome wide association. This collection will be helpful to identify agriculturally desirable traits from other supspecies.

Sorghum is the fifth most important cereal in the world as one of the staple food. For the use of natural dye, we have done some researches about sorghum red pigments extracted from stalk and leaves on its physiochemical properties, extracting methods and applications. The researches involved maximum extraction of sorghum pigment and analysis of its processing condition. Total polyphenol and tannin contents were measured by varieties and different plant parts. The stabilities of pigment by irradiation and heat treatment for processing were measured by colorimeter and thermal gravimetric analysis (TGA). In addition, hybrid nano-silica composites with sorghum pigment were made by combining with polyvinyl alcohol, polyvinyl acetate and sodium silicate. Water silica hybrids with sorghum pigment were performed by emulsion treatment. Nano-silica particles were identified and measured their size to be about 200 ~ 400 nm by SEM analysis.

In plant, senescence is associated with various aspects of the final stage of leaf development, nutrient relocation from leaves to reproducing seeds and stress resistance, and yield which is the most important trait in crops. Thus, the increase of knowledge on the regulatory processes of plant senescence will allow us to manipulate senescence for agronomic benefit in the future. of genetic studies have been conducted with mutants, where most of studies were focused on the delayed senescence mutants which are associated with positive factors on senescence by treating EMS to Koshikari, we induced a mutant showing early senescence phenotype, which possibly enable us to identify a negative factor of senescence. The appearance of the mutant is identical before booting stage and then the mutant showed senescence phenotype rignt before booting stage whereas Koshikari have health green leaves. The clumn length of the mutant is 98cm and the panicle length is 23cm as same as those of Koshikari. The chlorophyl contents of the mutant leaves, measured by SPAD, decreased during senescence. The soluble protein contents in the mutant leaves also decreased but no differences in the constitution reolved 1D-SDS-PAGE was detected. However, an additional shotgun proteomic approach to detect the differences of the protein constitutions during the senescence in the mutant leaves will be conducted.

Two sugary mutants, Hwacheong sugary-1 (Hsu1) and Hwacheong sugary-2 (Hsu2) were obtained by chemical mutagenesis from japonica cultivar, Hwacheongbyeo. Sugary mutants exhibited wrinkled and translucent grain with high soluble sugar content. In addition, amber-colored endosperm of sugary mutants was loosely packed due to abnormal starch granules compared to densely packed wild-type. Especially, the grain of Hsu2 mutant was less wrinkled than that of Hsu1, thus Hsu2 can be polished easily. Previous studies reveal that su1 mutant was resulted from mutation in gene for a debranching enzyme, isoamylase but the sequence of the mutated gene has not been identified. To identify the sequence of sugary genes, the map-based cloning strategy was applied. The genetic study revealed that the phenotype of Hsu2 mutant was controlled by two recessive genes. Interestingly, one of the genes was located on chromosome 8 at the position of isoamylase which was known as su-1. This indicates that mutation in isoamylase gene causes sugary endosperm characteristics. However we found different mutation points between the Hsu1 and Hsu2. The point mutation in Hsu1 was occurred at 10th exon whereas the other mutation related with Hsu2 was occurred at 15th exon. As mentioned above, the Hsu2 mutant has less wrinkled shape and less soluble sugar content than the Hsu1 mutant. Thus, we hypothesize that the other gene controlling Hsu2 mutant phenotype may have a role in weakening the effect of the su-1. Further study on the other gene associated with the Hsu2 phenotype is in progress.

Chinese cabbage (Brassica rapa ssp. pekinensis) is one of the most important vegetables and widely cultivated in Asia countries including Korea and China. Recently, whole genome sequence and full-length cDNA information of this species became available, which are encouraging genetic studies of this species to characterize agricultural important traits. Orange-colored (Or) cultivar of Chinese cabbage has inner leaves in orange, whereas other cultivars generally cultivated have yellow (Ye)- or white-colored inner leaves. In this study, we investigated phenotypes and carotenoid biosynthesis genes related to color variation in the Or cultivar. Firstly we compared the carotenoid content and composition between the Or and Ye cultivars by HPLC analysis. The inner leaves of Or cultivar contained approximately 9-fold high β-carotene content, whereas content of both lutein and violaxanthin was decreased to less than 30%, compared to Ye cultivar. Or cultivar was segregated with ratio of 3:1 in F2 population derived from crossing between Or and Ye inbred lines, indicating that Or phenotype is controlled by single recessive gene. To identify this gene, we investigated the expression of several genes involved in carotenoid biosynthesis by RT-PCR analysis. Among genes tested, two encoding putative carotenoid isomerase (CRTISO) and phytoene desaturase (PDS) were identified to show different expression between Or and Ye cultivars. Through further analysis of genomic DNA regions of these two genes, we could expect that several mutations such as InDel and base-substitution occurred and then affected expression of these genes in Or cultivar. In this presentation, I will introduce more detailed results for Or cultivars.

Japonica rice cultivars exhibit high susceptibility to bacterial blight(BB) disease due to genetic vulnerability in Korea. Korean japonica resistant rice cultivars mainly possess one of the genes, Xa1 or Xa3 for BB resistance. These resistance genes are becoming susceptible to K3a, resulting in serious rice yield reduction. This study was carried out to confirm the effect of xa13 gene pyramiding for developing of japonica rice cultivars resistant to BB pathogen breaking down Xa1 or Xa3. IRBB4 conferring Xa4 gene was resistant to K1(HB01013), K2(HB01014), K3(HB01015), and moderately resistant to K3a(HB01009). IRBB5 having xa5 gene was resistant to K1, K2, K3, and K3a. The recessive gene xa13 was resistant to K1 race but susceptible to K2, K3, and K3a. But Xa21 gene is susceptible to predominant K1 race but resistant to other races such as K2, K3, and K3a. Two genes Xa3 and xa13 were susceptible and Xa4 gene was moderate resistant to 24 isolates. xa5 and Xa21 genes were resistant to all isolates including K3a. When xa13 gene combined Xa4, xa5 and Xa21 genes, effect of xa13 gene pyramiding showed higher resistant reaction than that having singe gene out of Xa4, xa5, and Xa21. The order of resistance against 24 isolates breaking down Xa3 gene was IRBB55(xa13+Xa21) > IRBB53 (xa5+xa13) > IRBB51 (Xa4+xa13).

‘Jungmo2502’ awnless barley line (Hordeum vulgare L.) developed at National Institute of Crop Science, RDA in 2011. The Jungmo2502 derived from three-way cross between Millyang100 and F1 (Samheung/Suwon300) in 2000. Subsequent generation were handled by the bulk method in a pedigree selection program. A promising line showed high yield as well as lodging resistance in the yield trial tested at Iksan in 2007 to 2008, designated as Iksan 448. The line in the regional yield trials(RYT) tested in seven locations around Korea for three years from 2009 to 2011. The Jungmo2502 is carrying the growth habit of Ⅳ, green leaf and stem similar to the check cultivar ′Youngyang′. Its heading date was May 3, and maturing time was May 30 in paddy field, which was 3 days later respectively than check cultivar. It was 98 cm of culm length, 607 spikes per m2 and showed more rate of leaf blade, winter hardiness, and resistance to BaYMV than the check cultivar. Average dry matter yield was similar with the check cultivar in paddy field (about 11 ton ha-1). It also showed good qualities as 8.7% of crude protein, 21.9% of ADF (acid detergent fiber), 40.7% of NDF (neutral detergent fiber), and 71.6% of TDN (total digestible nutrients), including high grade of silage for whole crop barley.

Development of transgenic plant with desirable traits to cultivated plant is one of the important procedures in plant molecular breeding. However, applicable assessment of transgenic plant in laboratorial scale is not much except cultivating transgenic plant for a whole life in field condition. Here, we analyzed chlorophyll fluorescence in three transgenic soybean lines with AtMYB44 transcription factor for assessment of photosynthetic activity under abiotic stresses such as drought. Soybean varieties used in this study were ‘Bert’ and ‘Bert’ derived three transgenic soybeans, ‘AtMYB44 CM35101’, ‘AtMYB44 CM2471’, and ‘AtMYB44 CM4481’. Analyzed five different chlorophyll fluorescence variables are maximum PSII quantum yield (QY_max), steady state PSII quantum yield (QY_Lss), steady state non-photochemical quenching (NPQ_Lss), coefficient of photochemical quenching in steady-state (Qp_Lss), and fluorescence declineratio in steady-state (Rfd_Lss). To determine main chlorophyll fluorescence variable affected by abiotic stress, principal component analysis (PCA) was conducted with five chlorophyll fluorescence variables measured from four varieties. QY_Lss and NPQ_Lss were main chlorophyll fluorescence variables to evaluate abiotic stress, particularly in drought stress. In comparison with transgenic soybean lines based on chlorophyll fluorescence variables, ‘AtMYB44 CM2471’ and ‘AtMYB44 CM4481’ are more tolerant to drought than the others. Interestingly, three transgenic soybean lines which have a same AtMYB44 gene with different regions of chromosome revealed the quite different responses of chlorophyll fluorescence to drought treatment.

Pollen development in flowering plants is regulated by a comprehensive pattern of genes. One way to produce hybrid rice based on nuclear male sterility is to find out firstly the potential promoters that function specifically in anthers since it is a specific site for transcription initiation and play key roles for the spatial and temporal expression of the genes. To implement this objective, we were selected promoter region of 16 genes based on the expression pattern of microarray and then those were introduced into the promoterless final destination vector which containing the GFP and GUS reporters genes. The resulting twelve vectors were transformed into monocotyledonous rice (Oryza sativa L) and a dicotyledonous Arabidopsis as heterologous system. Minimum 20 plants for each vector were analyzed by histochemical GUS assay at the flowering stage in Arabidopsis. 9 vectors out of 12 vectors constructed were expressed exclusively at the anther, especially in pollen, however one vector exhibited expression in stigma. For rice, T-DNA insertion were confirmed with specific primers in each promoter and GFP region. All T0 transgenic plants contained T-DNA insertion in their genome. This study would provide valuable information for biotechnological application for the induction of male sterility in plants.

Mitogen-activated protein kinase (MAPK) signaling cascades play critical roles in various cellular events including abiotic/biotic stress responses, innate immunity, hormone signaling and cell specificity in plants. The MAPK-mediated stress and ethylene signaling are recently known to be involved in nitogen-fixing symbiotic interactions; however, the biological role of MAPK for nodule development in legume plants is largely unknown. We here elucidated that MtMKK5-MtMPK3/6 cascade negatively regulate the nitrogen fixing nodule formation in Medicago truncatula. MtMKK5, an ortholog of SIMKK, overexpression significantly reduces the nodule formation in M. truncatula roots. MtMKK5 directly activates MtMPK3/6 by phosphorylation on the TEY motif within the activation loop in the cytoplasm, which might link to EFD as a negative regulator for nodule formation. EFD has a putative MAPK phosphorylation Thr residue and could be a target of the activated MtMPK3/6 in the nucleus. Consistently, a MAPK specific inhibitor U0126 enhances nodule formation and confers similar nodule phenotypes to the efd-1 mutant such as lower proliferation and differentiation to symbiotic tissues. Our works thus reveal a key negative signaling module mediated by MtMKK5-MtMPK3/6-EFD for symbiotic nitrogen fixing nodule organogenesis.

Grain size is one of the most important trait determining yield in cereal crops, apart from number of grains per panicle, number of panicles per plant and 1000 seed weight. Other than grain characteristics, plant architecture is another very important factor influencing yield by affecting the amount plant surface area directly exposed to the sun light. Erect panicle is important morphological characteristic which helps in enhancing the yield by allowing sun light to fall directly on leaves unlike curved panicle which blocks sunlight and consequently reduce photosynthesis. A small round grain and erect panicle mutant was obtained by treating Hwacheong rice (japonica) with MNU (N-methyl-N-nitrosourea) chemical mutagen. Through bulked segregant analysis (BSA) using STS (Sequence-Tagged Sites) and SS-STS (Sub-species Specific Sequence-Tagged Site) markers we located the mutated gene on the long arm of chromosome 7 and narrowed down candidate region to 168.75kbp through fine mapping. Mutant manifested characteristics like reduced grain size and plant height, dense and erect panicle and relatively erect plant compared to the wild type. When we crossed the mutant with its parent (Hwacheong), F1 panicle and grain characteristics showed intermediate phenotype, therefore, we concluded that wild type allele of this gene shows incomplete dominance. Scanning electron microscopy(SEM) result shows that increase in width of mutant grain, which changes its shape, is due to increase in width of glume cells. Phenotypic examination shows that dense and erect panicle phenotype is result of reduction in length of rachis, primary and secondary branch.

Amylose content of rice endosperm is one of the determinants of rice eating quality. This study was conducted to elucidate the mode of inheritance of dull gene in Milyang262, tentatively designated as du7(t), and to identify the molecular marker for du7(t) to be employed in marker-assisted breeding and gene pyramiding. Genetic analysis was carried out on F2 population derived from a cross between Junam and Milyang262. The low amylose content of Milyang262 was indicated to be under single recessive control. Allelism tests were as well conducted by crossing Milyang262 with Baegjinju and Baegokchal, which harbor du1 and wx gene, respectively. du7(t) was demonstrated to be inherited independently to du1 and wx. F2 population of Baegokchal/Milyang262 was used for molecular mapping. Linkage analysis was conducted on a population consisted of 120 individuals by several SSR markers. Initial mapping indicated that du7(t) is located on the end of long arm of chromosome 6 between SSR marker RM20590 and RM3509. To fine map the gene, a bigger population and several additional markers were employed. du7(t) was further mapped to a 1.74 Mb region between two SSR markers (RM6926 and RM412). Furthermore, we indentified three SSR markers that co-segregated with du(t) i.e. RM6811, RM3765, and RM176.

The early senescence mutant was isolated from the japonica rice Koshihikari through Ethyl-methane-sulfonate(EMS) mutagenesis. The early senescence phenotype was controlled by a single recessive nuclear gene, tentatively symbolized as es-k. Using phenotypic and molecular markers, we mapped the es-k locus to the long arm of chromosome 7 between STS markers 147-1 and 150-1, a physical region of 370-kb. The symptom of early senescence appeared even before heading, while appeared after heading in those of the wild-type rice during senescence. Early stage physiological characteristics of the es-k mutant was similar to that of the wild-type. However, after heading, es-k mutants started to exhibit a significant decrease in chlorophyll content, soluble protein content 10 days earlier compared to normal type. To characterize the interaction with the environment factors, experiments were carried out under controlled temperature and light conditions, respectively. The wild-type leaf color appeared normal irrespective of temperature treatment, while the leaf of es-k mutant appeared pale-green at the low temperature and dark-green at the high temperature. During dark-induced senescence, mutant did not show significant differences compared to normal type. The results show that es-k is sensitive to temperature but not to light.

"United Nations Convention on Contracts for the International Carriage of Goods Wholly or Partly by Sea" was passed in July 3, 2008. on September 23, 2009 signing ceremony was held in Rotterdam. The system of maritime performing party is a special system in the transport convention, and constitutes an integral part of the Convention. Maritime performing party system is not the first system which brings in the carrier's independent contractor, but it is the most comprehensive and thorough one. It unified the duty of carrier's independent contractor in the maritime segment, and it is also an important progress in the developing process of international maritime legal system. There are some differences between the maritime performing party and China's current related system, i.e, the port maritime performing party and the intermediate performing party are included in the maritime performing party, and they can enjoy the carrier's exception clause and limitation of liability.

Working onboard large ocean-going vessels is one of many industries that is by its very nature extremely error-provoking. In industries such as aviation, nuclear power, healthcare and transport, the need for mental preparedness and professionalism are amongst the qualities essential for the management of complex hazardous systems. Recruiting, training, and more importantly retaining the right people to work in these industries is of paramount importance if safety is to be continuously improved and economic benefits realized. Through consideration of extant empirical literature this paper will explore the need for enhancing collaboration between shipping companies and maritime training insitutes in order to provide the industry with those most capable for the job. It will be concluded that by providing a strong maritime foundation from the outset of training, maritime academies in South Korea can provide shipping companies with a wealth of highly qualified, motivated officers who will contribute to safety and remain valuable assets within the industry for years to come.