Slaughter of cattle, pigs, and chickens has increased continuously. In particular, slaughter of chickens has been grown up about 150% in 2010 than that in 2003, that is approximately 120,000 tons. All of them are underwent consigned treatment even though those can be used as a resource and an energy source. With this regards, THR (Thermal Hydrolysis Reaction) leads to reduce water content drastically (<30% in sludge cakes). In addition, Dehydrated solid would be re-used as solid fuels (SRF) as well. In this study, We have applied THR to a plant (10 ton/day) on the basis of our lab and pilot results. Water content of sludge cakes showed with a ranges of 30 to 40% after solid-liquid separation. Dairy SRF produced 1.5 ton/day and its heat capacity for SRF has 6,500 kcal/kg. This gave the steam produced about 12 ton/day throughout the plant operation, suggesting that THR system would expect energy savings.

Recently, the importance of food safety is increasing due to numerous junk food. Junk food means to violate the law in stage such as production, manufacture, distribution, and sale of food. Many crop plants are processing as foods including bread, noodle, and other foods for supporting nutrition to human. For example, rice is one of the most well-known food crops in the world, and processed rice is being mixed with other processed crops to health food. The object of this study is to detect amount of specific grains, i.e. rice from processed foods mixed with other cereals. This experiment was performed to the following two steps: 1) designed the specific primer sets based on chloroplast DNAs, 2) amplified products using real-time PCR. We designed eleven primer sets within chloroplast DNA of rice, and then the confirmation of primer efficiency was to amplified with rice genomic DNA using real-time PCR. In addition, these primer sets were applied in other crops such as wheat, maize, and adlay to confirm specificity to rice. The rice specific primer sets were determined by the number of amplification and the melting peak through real-time PCR. As a result, five primer sets were confirmed to uniqueness in the rice genome. In conclusion, the specific primer sets would be useful for identifying rice grain from the processed foods to eliminate junk foods and for contribution of food safety.

Brassinosteroids (BRs) play important roles in many aspects of plant growth and development. BR-induced AtBEE3 (brassinosteroid enhanced expression 3) is required for a proper BR response in Arabidopsis. Here, we identified a poplar (Populus alba x P. glandulosa) BEE3 homolog encoding a putative basic helix-loop-helix (bHLH)-type transcription factor through microarray analysis. Transcripts of PagBEE3 were mainly detected in stems, with the internode having a low level of the transcripts and the node having a relatively higher level. The function of the PagBEE3 gene was investigated through the phenotypic analyses with PagBEE3-overexpressing (ox) transgenic lines. This work mainly focused on a potential role of PagBEE3 in stem growth and development of polar. The PagBEE3-ox poplar showed thicker and longer stems than wild-type plants. The xylem cells from the stems of PagBEE3-ox plants revealed remarkably enhanced proliferation, resulting in an earlier thickening growth than wild-type plants. Microarray analysis revealed that the expression of many genes involved in xylem cell proliferation and development was altered in the PagBEE3-ox plants. Therefore, this work suggests that xylem development of poplar is accelerated in PagBEE3-ox plants and PagBEE3 plays a role in the stem growth by increasing the proliferation of xylem cells to promote the initial thickening growth of poplar stems.

Rice is a staple food for over one-half of the world population, especially in Asian countries. Recently, the growth and yield of crop plants was affected by various abiotic stresses, such as salt, drought, and high temperature due to change of climate environment. To study molecular functions of Oryza sativa nuclear-targeted RING Finger Proteins (OsNRFPs) in response to abiotic stresses, we selected 44 OsNRFPs genes, whose subcelluar localizations are predicted to the nucluear, on the basis of expression patterns of a microarray dataset. A total of 44 OsNRFPs were grouped into two types such as RING-HC and RING-H2 via phylogeneitc analysis of their RING domains structures. Subsequently, we surveyed the expression patterns of 44 genes in response to salt stress via qRT-PCR in roots. We found 10 salt stress-induced OsNRFPs and then examined their subcellular localizations. These genes were clearly localized to the nucleus (OsNRFPHC-10), cytoplasm (OsNRFPHC-17 and OsNRFPH2-16) and microtuble (OsNRFPHC-23, OsNRFPH2-17 and OsNRFPH2-05), respectively. These results might provide a key clue for understanding moleuclar functions of the OsNRFP genes associated with salt stress-related signaling pathway

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

Chrysanthemum white rust, caused by Puccinia horiana, is one of the most destructive fungal diseases in chrysanthemum cultivation worldwide. For increasing efficiency of resistant breeding, molecular markers linked to chrysanthemum white rust resistance gene were developed in pseudo F1 cross population between ‘Puma White’ as susceptible and ‘Dancer’ as resistant using bulked segregant analysis (BSA). Of 280 RAPD primers (Operon 10 mer), 18 primers found to be polymorphic. After screening of these primers in 20 individual lines, only OPI-13520 was selected as closely linked marker to white rust disease resistance. Based on correspondence between phenotypic resistant level and marker in 187 segregation population, the genetic distance between white rust resistance gene and OPI-13520 marker assumed to be 3.8 cM. For OPI-13520 marker conversion into sequence characterized amplified region (SCAR) marker, the amplified fragment of OPI-13520 was purified, cloned and sequenced. Based on the DNA sequence of OPI-13520, SCAR maker was generated and verified in 20 individual lines used in BSA-RAPD.The results showed SCAR marker could be used to identify white rust resistance in chrysanthemum.

The metalloid arsenic (As) and the hevy metal cadmium (Cd) are ubiquitously found at low concentrations in the earth, while high concentrations of the both elements in soil and crop are severe dangerous to human health. We have tried to retrieve RING E3 ligase gene, which is believed to regulate substrate proteins in As or Cd uptake via ubiquitin 26S proteasome pathway, related to inhibit metal ion transport system. A total of 48 rice RING E3 ligases were randomly selected and then conducted semi-quantitative RT-PCR for their expression patterns as exposed to As and Cd treatments. We discovered one gene, Oryza sativa heavy metal induced RING E3 ligase 1 (OsHIR1) that was significantly up-regulated against both treatments. A total of 31 positive interaction clones with OsHIR1 were screened depending on their strong α-galactosidase activity via yeast-two hybrid screen. Bimolecular fluorescence complementation analysis evidenced that the OsHIR1 protein was clearly interacted with each of six partner protein including aquaporin tonoplast intrinsic protein 4;1 (OsTIP4;1) in the plasma membrane. Protein degradation assay showed that OsHIR1 strongly degraded the protein level of OsTIP4;1 via ubiquitin 26S proteasome system. Heterogeneous overexpression of OsHIR1 in Arabidopsis showed As- and Cd-insensitive phenotype. In addition, the transgenic plant showed low levels of As and Cd accumulation than the control plant in leaf and root. Here, we report the novel finding that OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptake.

Plant growth under water-deficit conditions adversely affects many key processes. Efforts to understand drought stress-related defense mechanisms have revealed a host of plant genes using molecular approaches in rice. Here, we report the novel finding that OsCTR1 E3 ligase regulates both chloroplast-localized chloroplast protein 12 (OsCP12) and ribosomal protein 1 (OsRP1) in protein levels and subcellular localization. The results of a yeast-two hybrid assay, bimolecular fluorescence complementation assay, ubiquitination assay, subcellular localization, and a protein degradation assay support the hypothesis that OsCTR1 functions in trafficking inhibition and proteolysis of OsCP12 and OsRP1 via the ubiquitin 26S proteasome pathway. Heterogeneous overexpression of OsCTR1 in Arabidopsis showed ABA-hypersensitive phenotype in seed germination, seedling growth, and stomatal closure. The transgenic plants also exhibited improvement of water-deficit tolerance with an accumulation of hydrogen peroxide production. These results demonstrate that the OsCTR1 E3 ligase might positively regulate the cellular functions of OsCP12 and RP1 related to photosynthesis under drought stress conditions in rice.

Low temperature is a major factor restrict to growth and limiting productivity of rice crops. We used a cDNA microarray approach to monitor the expression profile of rice (Oryza sativa) under chilling stress and identified 20 chilling inducible genes in previously study. Ten such genes encoding bHLH, metal transporter and, zinc finger protein with unknown functions showed a significant change in expression under various abiotic stresses. Among them, OsCHI1 (Os07g15460), OsCHI2 (Os02g43660), and OsCHI3 (Os01g61160), were selected for further study. They have structural features such as metal-binding signature sequences in their protein sequences, and OsCHI genes were expressed in root of rice seedling and induced in chilling and salt or drought. Expression of OsCHI1, OsCHI3 and OsCHI2 were targeted to membrane and ER when transiently expressed in tobacco cell, respectively. The Arabidopsis (Arabidopsis thaliana) transgenic plants overexpressing showed increased tolerance to salt and drought stress in the seed germination and root elongation than that of wild type. This comprehensive study provides insight into the biological function of OsCHIs, which may be useful in understanding how rice plants adapt to unfavorable environmental conditions.

We used a microarray dataset that is deposited in the public database to evaluate plant responses to heat stress and selected two genes, OsSHSP1 (Os03g16030) and OsSHSP2 (Os01g04380), that are highly expressed under heat stress in rice. OsSHSP1 and OsSHSP2 gene transcripts were highly induced in response to salt and drought. In addition, OsSHSP1 and OsSHSP2 gene transcripts were induced under ABA and SA. Subcellular localization of proteins of 35S::OsSHSP1 were associated with the cytosol, whereas those of and 35S::OsSHSP2 were associated with the cytosol and nucleus. Heterogeneous overexpression of both genes exhibited higher germination rates than those of wild-type plants under the salt treatment, but not under heat or drought stress. The network of both genes harboring 9 sHSPs as well as at least 13 other chaperone genes might support the idea of a role for sHSPs in the chaperone network. Our findings might provide clues to shed light on the molecular functions of OsSHSP1 and OsSHSP2 in response to abiotic stresses, especially heat stress.

In order to better understand the biological systems that are affected in response to cosmic ray, we conducted the weighted gene co-expression network analysis with module detection method. By using the Pearson’s correlation coefficient value, we were evaluated the complex gene-gene functional interactions between 680 CR-response probes from integrated microarray datasets, which included large-scale transcriptional profiling of 918 microarray samples. These probes were divided into 6 distinct modules that contained 20 enriched function such as oxidoreductase activity, response to stimulus and stress, and hydrolase activity. Especially, module 1 and 2 commonly showed the enriched annotation categories such as oxidoreductase activity, including the enriched cis-regulatory elements known as ROS specific regulator. These results suggest in module1 and 2 that ROS-mediated irradiation response pathway are affected by CR. We found the 243 irradiation-dependent probes, which were exhibited the similarities of differentially expressed patterns in various irradiation microarray datasets, and RT-PCR for confirmations of several irradiation-dependent genes were exhibited the similar expressed patterns in rice by CR, gamma ray and Ion beam treatments. Interestingly, these genes were differentially expressed by non-gravity. Moreover, we were identified the co-regulations between several irradiation-dependent genes and functional interacted genes in the CR-responsive network by various GA treatments such as different conditions of dose and treatment time. These results of network-based analysis might provide a clue to understanding the complex biological system of CR.

Rice is one of the most important food crops in the world, and has been used as model monocots for genetic studies, because of its relatively small genome size. We have previously reported the different functions of several RING (Really Interesting New Gene) proteins to respond the various abiotic stresses. In order to study a regulation of RING proteins in rice under ionizing irradiation such as gamma ray (GA), we have identified the expression patterns of these genes by RT-PCR. We found Gamma-ray induced RING finger protein (OsGRP) gene, which were associated with cytosol by subcellular localization analysis. in vitro ubiquitination assay revealed that OsGRP possess E3 ligase activity. Also, we demonstrate that C196A point mutation in the RING finger domain of OsGRP can have a critical effect to the breakdown of structural integrity in RING constructs. To identify the interaction partners for OsGRP in protein-protein interactions, we found the seven genes interacted with OsGRP by Yeast Two Hybrid method. To examine the GA-influence of interaction partners by RT-PCR, two genes were specifically down-regulated in rice during GA treatment. These interaction partners were identified the reliable interactions and subcellular localizations via BiFC method. Interestingly, five genes associated with plastid, while two down-regulated genes associated with cytosol and plastid. These results of OsGRP based on genetic approach might provide a clue to understanding the GA responsive mechanism in rice.

Abscission is an important developmental process used to shed organs such as leaves, flowers and fruits. Despite the detailed characterization of growth dynamics and hormonal balance during the early steps of fruit development, the molecular aspects remain unclear. Abscission of young fruit occurs by separation of cells in anatomically distinct regions between the pedicel and junction. Differences of gene expression between central pedicel and lateral pedicel were investigated by NGS. Partial cDNAs from 15 clones from both the central pedicel and lateral pedicel were selected for nucleotide sequence determination and homology searches, and 12 clones were subsequently selected for further analysis. In preliminary series of Real Time PCR analysis, 9 genes were confirmed as showing a higher expression level in lateral pedicel than in central pedicel. Many of these genes are expressed in a central or lateral pedicel in specific manner, and the expression profiles of the representative genes were confirmed. To clarify the mechanism of MdIAA14 transcription factor gene underlying abscission zone development, we are investigating the expression patterns between central and lateral pedicels in different apple cultivar using real-time PCR and constructing the vector for transformation into apple.

Inactivation of the gene (DFR-A) coding for dihydroflavonol 4-reductase (DFR) involved in the anthocyanin biosynthesis pathway results in a yellow bulb color in onion (Allium cepa L.) and three inactive alleles have previously been identified in onion. Additionally, three active and six inactive DFR-A alleles were newly identified from extensive analyses of diverse onion germplasm. Presently, a yellow mutant containing a 171-bp deletion in the promoter region was identified and designated DFR-APD. Critically reduced transcription of this mutant allele and perfect co-segregation with color phenotypes in segregating populations were observed. Another yellow mutant (DFR-A5’DEL) containing a 518-bp deletion covering exons 1 and 2, which played important roles in DFR function, was identified. Meanwhile, both 2-bp and 4-bp insertions in the coding region leading to creation of pre-mature stop codons were also identified and designated DFR-AGT and DFR-A2AT, respectively. A 1-bp substitution mutation (DFR-AK48N) changing a positively charged lysine residue into a neutral asparagine was identified. This lysine residue, a NADPH binding site, was strictly conserved in other species. In addition, insertion of a leucine residue around substrate binding sites and catalytic triad was identified in several yellow accessions and was designated DFR-ATTA. Phylogenetic analysis of DFR-A alleles showed that all inactive alleles were independently derived from four different active alleles. In addition, the close relatedness and diversity of DFR-A mutants implied that all these mutations might have occurred after domestication of onions and had probably been maintained by artificial selection.

A 72-year-old female patient was admitted to our hospital with a rapidly growing left eyelid mass. Other ocular findings were normal. Excision biopsy and a cytological study were performed. Histopathology of the excised tissue showed malignant cells in extracellular lumens and glands with prominent micronuclei and mitotic figures. The tissue was also strongly positive for cytokeratin. As a result, of these fingings primary eyelid hidradenocarcinoma was diagnosed. This case demonstrates that hidradenocarcinoma should be considered in the differential diagnosis of primary eyelid tumors.