Gene reg비 at i o n during the human craniofacial development is not well understood In effort to understand n ewly identifï ed genes that may play role(s) in the human craniofacial development, non-redundan t genes were isolated from the s ubtracted cDNA libra ry of human embryonal craniofacial tissues and examined their possible structu ral rolc in parallcl with thosc gcncs from isolatecl human c h o nclroc)πes cDNA library. Fifty genes were init ia ll y chosen from 398 clones iso latecl were used for selective dominant expression in both chondrocytes and the craniofacial sections of 10 weeks old human embryo by in situ hybridization method. Based upon the high levels 。f expression, we have identifi ecl seven unknown genes; ch89, ch96. ch129. ch 153. ch 276 ch285. and ch334 . In 。rder to unde rs tancl the possi ble role of these genes‘ the structural simulation of the expressed proteins were constructecl by Sybyl 6.6 program. Ch 276 gene was same with a clone, c14 0 1' f173. registered in GenBank(NM_022489) a nd is composed 0 1' 323 amino acids having a reverse s ignaling domain from the extra- cellular matrix(C-terminal) to cell membrane(N-terminal) and 12 turns of helical structure. Gene protein also r etains a famil iar fïbronectin binding domain(RGD). three s ites 0 1' Ca ion binding motifs. cAMP- and cGMP-dep endent protein kinase phos phorylation site, two regions of protein kinase C phosphorylation s ites. glyco- saminoglycan attachment s ite ancl N-glycosylation site. transmembrane and Al kaline Phosphatase active s ite domains This newly iclentifï ed human protein from human choncl rocytes cDNA library appearecl to be related to a known calcification s ignaling protein. was named as Ca lsin(Ch276) . Ch153 appeared to be related a family of anti-microbial peptide acting as an inflammation mediator and Ch334 clone as a zinc finger protein whose expression in creases in human adult ti ssue‘ These results suggest that these novel genes ident i!ï ed from human chondrocytes rnay provide a new path 0 1' embryonic cartilage development and human craniofacial development.

Mannitol is commonly used to reduce intracranial and intraocular pressures and to prevent dialysis-disequilibrium syndrome. However, intravenous mannitol infusion in various cases has the potential to result in acute kidney injury (AKI). We present a case of mannitol-induced AKI that developed after low dose mannitol infusion and resulted in recovery after hemodialysis. A 66-year-old woman was admitted to the hospital with a diagnosis of left middle cerebral artery infarction. On hospital day 5, cerebral edema was observed on a follow-up MRI. D-mannitol 35 g was given intravenously every 8 hours. Four days later, serum creatinine levels were elevated from 1.2 mg/dL to 3.5 mg/dL. The serum osmolal gap was found to be 52.4 mosm/kg H2O and urine output was reduced from 2.78 mL/kg/h to 0.69 mL/kg/h over three days. Hemodialysis over 2 hours was performed and renal function subsequently improved to baseline function. A potential risk of AKI exists even with low dose mannitol infusion in patients with advanced age, underlying renal impairment, and concomitant use of nephrotoxic agents. Mannitol-induced AKI may be rapidly reversed by short-term hemodialysis.

Foxtail millet (Setaria italica L.) is the second most widely cultivated species of millet, especially in East Asia and is a tractable experimental model crop for studying functional genomics of millets. However, insufficient researches had been conducted about the foxtail millet germplasm and is significantly impeding its genetic improvement. We attempted to develop EST-derived-SSR (eSSR) markers and utilize them in genetic comparison of germplasm and transferability. A total of 66,027 foxtail millet EST sequences and 42,754 genomic sequence were deduced transcriptom. Approximately 42,000 single tone contigs were generated using DNAstar 5.0 software for redundancy minimization. Nearly 33% of the 14,012 unigenes contained SSRs, but primers were designed for a total of 314 microsatellites concentrating with more than 24 bp of repeats. A total of 314 primers were successfully designed with more than 24 bp of repeats. From these microsatellites, 56 primer pairs were showed polymorphism with over than 15 bp differences among 96 accessions collected from different countries. Polymorphic information content (PIC) value ranged from 0.020 to 0.700 with an average of 0.381 indicating moderate level of informativeness within these EST-SSRs markers. The EST-SSR markers developed in this study will serve as a useful source for genetic studies, such as genetic variability, transferability, association mapping, and molecular breeding

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.

Rice blast (Magnaporthe oryza B.) is one of the most widespread and devastating diseases of rice. Screening of valuable genetic resources harboring resistance genes is one of the most efficient approaches against blast disease. Because the bioassay to rice blast in the field shows high variations, this study has performed to provide DNA profiles in the accessions of diverse countries using major blast resistant genes linked markers, identified and mapped in different genotypes of rice. Because durable resistance to blast is controlled by a combination of major resistance genes, we surveyed the distribution of blast resistant genes in the 1,500 accessions using major 12 blast resistance genes linked markers. These resistant genes found that the frequency distribution of Pi-39 (66.9%), Pik-m (41.9%), Pit (40.5%), Pii (21%), Pib (19.3%), Pi-d(t)2 (12.7%), but Pita, Pita/Pita-2, Pik, Piz-t, Pi5 genes were identified in less than 10% frequency. Most of accessions contain from 1 to 4 different resistant genes. Pi39 and Pik-m genes amplified in the 69.1% and 51.7% among 356 Korean accessions, Pi39 (79.6%) and Pib (55.8%) in 113 China, Pit (80.6%) and Pib (32%) in 103 Philippines, respectively. In this study, we evaluated the blast resistance degree and the information about the distribution of rice blast resistant genes in rice germplasm. This study will help to develop effective strategies for managing rice blast disease in rice germplasm.

This study is a basic study on the development of functional substances involved in obesity prevention, lipid metabolism, and immune regulation. Male Sprague-Dawley rats were fed a high-fat diet for 10 weeks. Allium monanthum extracts (AME) were administered orally to obesity-induced rats, and their lipid-lowering, antioxidative and various types of biological effects related to the immune system were examined. Blood free fatty acid and triglyceride concentrations decreased as the dose of AME increased. Total cholesterol and LDL cholesterol concentrations in the blood decreased as the dose of AME increased. The total cholesterol concentrations in the liver of the AME-treated groups were lower than the control group. The thiobarbituric acid reactive concentrations were lower in the plasma and liver of all AME-treated groups than the control group. Plasma AST and ALT activities did not show any significant differences among the treatment groups. IL-1β and IL-6 concentrations in the liver tended to decrease as the dose of AME increased. TNF-α and IL-10 concentrations did now show any significant differences compared to the control group. Lower expression levels of TNF-α, Apo-B and Apo-E genes were found in the AME-treated groups. Taken together, these results indicate that AME may show positive effects in lipid lowering, antioxidation and anti-inflammation.

R genes are a key component of genetic interactions between plants and biotrophic bacteria and are known to regulate resistance against bacterial invasion. The most common R proteins contain a nucleotide-binding site and a leucine-rich repeat (NBS-LRR) domain. Some soybean NBS-LRR genes have also been reported to function in disease resistance. A total of 319 genes were determined to be putative NBS-LRR genes in the soybean genome. The number of NBS-LRR genes on each chromosome was highly correlated with the number of disease resistance QTL in the 2-Mb flanking regions of NBS-LRR genes. In addition, the recently duplicated regions contained duplicated NBS-LRR genes and duplicated disease resistance QTL, and possessed either an uneven or even number of NBS-LRR genes on each side. The significant difference in NBS-LRR gene expression between a resistant near-isogenic line (NIL) and a susceptible NIL after inoculation of Xanthomonas axonopodis pv. glycines supports the conjecture that NBS-LRR genes have disease resistance functions in the soybean genome. The number of NBS-LRR genes and disease resistance QTL in the 2-Mb flanking regions of each chromosome was significantly correlated, and several recently duplicated regions that contain NBS-LRR genes harbored disease resistance QTL for both sides. In addition, NBS-LRR gene expression was significantly different between the BLP-resistant NIL and the BLP-susceptible NIL in response to bacterial infection. From these observations, NBS-LRR genes are suggested to contribute to disease resistance in soybean. Moreover, we propose models for how NBS-LRR genes were duplicated, and apply Ks values for each NBS-LRR gene cluster.

The genus Rubus belongs to the Rosaceae family and is comprised of 600-800 species distributed worldwide. Understanding the genetic relationships and genetic structure in Rubus species is important for enabling efficient management, conservation, characterization and utilization of the species. However, as a minor crop, genetic research foundation was limited to explore genetic diversity and relationships in Rubus species. The present study shows the results of application SSR markers that were developed from SSR-enriched libraries of the one Rubus species (Rubus coreanus Mique.) in our previous study. We used 34 polymorphic microsatellite markers to analysis of genetic diversity within the Rubus species, including redraspberry, blackraspberry, blackberry and mountainberry. All the 34 SSR primers pairs produced 483 polymorphic and reproducible amplification fragments. The largest number of alleles per primer pair was confirmed at GB-RC-167, GB-RC-100, GB-RC-076 and GB-RC-245, which contained 26, 25, 23 and 21, respectively. An average value of polymorphic information contents (PIC) were 0.74 with a range of 0.36 to 0.92. Population structure and phylogenetic analyses showed that all Rubus species formed three largely distinct clusters, which were confirmed by principal coordinate analysis (PCoA). We obtained the results that the developed SSR markers showed a substantial degree of genetic diversity in the various Rubus species distributed in Korea.

Recent release of whole genome draft sequences in legume species have led comparative genome studies among legume plants including Glycine max, G. soja, Cajanus cajan and Medicago truncatula. The majority of comparative genomic researches have been conducted based on synteny of coding sequences and coding sequence variations may be one of major forces for speciation and evolution. However, non-coding sequences have been also reported to be important phenotypic regulators. Especially, since short sequence motifs in the promoter regions are highly conserved, they are suggested to be another resources of interests in comparative studies. In this study, we predicted the conserved short sequence motifs by BLASTN algorithm using dicot promoter database from Softberry (http://www.softberry.com). A total of 37,396 conserved short sequence motifs were identified onto 2 kb upstreams of 46,367 high confident gene model of G. max (cv. Williams 82). Meanwhile, whole genome of 7 soybean landraces (G. max) and 7 wild soybean genotypes (G. soja) were sequenced at low depth of less than ten using Illumina Hiseq 2000. Among these genotypes, nucleotide variations were identified in predicted conserved regulatory motifs by mapping of short reads to the reference genome sequence using the Samtools program (http://samtools.sourceforge.net/). Fifteen and two genes, which have SNPs in regulatory motifs and no SNP in coding sequence, were identified by comparisons of inter-species and intra-species, respectively. qRT-PCR experiments are in progress for investigating differences of these 17 genes expressions at transcriptional level.

As soybean (Glycine max) is known for its high nutritional value of oil and protein, soybean has been domesticated and cultivated by one specific character trait based on human selection. Importantly, tracing back in time where G. max and G. soja, the undomesticated ancestor of G. max have diverged plays an important role in studying of genetic diversity and in investigating the common ancestor of soybean. In this study, we sequenced 6 G. max and 6 G. soja using Illumina’s Hiseq 2000 with a low coverage sequencing technology to estimate the divergence of times between genotypes and populations. A total of the 12 genotypes were sequenced at the average depth of 6.5 and resulted 892.5 Mb and 903.3 MB consensus sequences with the coverage of 91.54% and 92.65% for G. max and G. soja, respectively. The whole genome SNP analysis showed that G. max had lower frequency levels of polymorphism (~0.1%) than G. soja (~0.25%). And, a high number of SNPs located in introns were found among 6 G. soja genotypes as SNPs were approximately twice than those found in 6 G max. The number of SNPs in G. max intronic regions was 53,134, whereas a total of 133,329 SNPs were discovered in G. soja introns. Almost an equal number of SNPs were discovered in 5’ UTR and exon regions; however, different numbers of SNP in CDS and 3′ UTR were identified. By the rate of nonsynonymous change, divergence of time between G. soja and G. max would be investigated.

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.