The change of phenolic compounds and radical scavenging activity of black soybean flours after germination and roasting treatment were evaluated as a part of the purpose of setting the quality of black soybean flours for each application. The moisture content of roasted black soybean flours decreased significantly according to roasting temperature and time, and the crude ash, protein, and fat contents increased. Water binding capacity of roasted black soybean flours without and with germination increased significantly according to roasting temperature and time, however water solubility index and swelling power decreased. The lightness of roasted black soybean flours was significantly decreased, and the redness and yellowness increased. The phenolic compounds and radical scavenging activity of roasted black soybean flours increase with increasing roasting temperature and time. The total polyphenol contents of roasted black soybean flours without and with germination were 5.43∼7.81 and 4.52∼6.17 mg GAE/g, and total flavonoid contents were 2.90∼3.50 and 2.34∼3.01 mg CE/g, respectively. DPPH radical scavenging activity of roasted black soybean flours without and with germination was 254.98∼415.05 and 171.95∼295.15 mg TE/100 g, and ABTS radical scavenging activity was 459.74∼596.37 and 422.95∼526.85 mg TE/100 g, respectively. As a result, it is necessary to set quality standards for each application considering the quality and antioxidant properties of roasted black soybean flours.

Lentinus edodes is a popular edible mushroom in South-East Asia. This study was initiated to evaluate the antioxidant activities, tyrosinase inhibitory effects on the fruiting bodies of L. edodes extracted with acetone, methanol and hot water. The antioxidant activities were performed on β-carotene-linoleic acid, reducing power, 1,1-diphenyl-2-picrylhydrazyl free radical scavenging, and ferrous chelating abilities. In addition to this, phenolic acid and flavonoids contents were also analyzed. Methanolic extract of L. edodes showed the strongest β-carotene-linoleic acid inhibition as compare to others extracts. At 8 mg/ml, hot water extract showed a high reducing power of 0.96. The scavenging effects on 1,1-diphenyl-2-picrylhydrazyl radicals, acetonic extract was effective than other extracts. The strongest chelating effect (86.45%) was obtained from the acetonic extract at 1.0 mg/ml concentration. Antioxidant activities of the extracts from the fruiting bodies of L. edodes were increased with the increasing concentration. After application of reverse phase high performance liquid chromatography, coupled to a diode array detector and electrospray ionisation mass spectra, four phenolic compounds namely, naringenin, hesperetin, formononetin and biochanin were identified from acetonic extract. Tyrosinase inhibition of acetonic, methanolic, and hot water extracts of L. edodes were increased with the increasing of concentration. Results revealed that acetonic and methanolic extracts showed good, while hot water showed moderate activities of the tyrosinase inhibition at the concentration tested. This study suggests that fruiting bodies of L. edodes can potentially be used as a readily accessible source of natural antioxidants.

Reparative dentine formation requires newly differentiated odontoblast-like cells. Therefore, identification of the molecule that stimulates the odontogenic differentiation of precursor cells in the tooth pulp will be helpful for the development of strategies to repair damaged pulp. In this study, we examined the effect of N-acetylcysteine (NAC) on the odontogenic differentiation of MDPC-23 cells, a mouse odontoblast-like cell line derived from dental papilla, and primary cultured rat dental papilla cells (RDPCs). NAC (1-30 mM) suppressed production of reactive oxygen species in MDPC-23 cells in a dose-dependent manner. Although 5 to 20 mM NAC did not alter MDPC-23 cell proliferation, 1 or 30 mM NAC significantly inhibited it. NAC enhanced mineralized nodule formation and the expression of several odontoblast differentiation-associated genes in both RDPCs and MDPC-23. This NAC stimulatory effect was significant, even at concentrations lower than 1 mM. However, NAC did not stimulate expression of bone morphogenetic protein-2, -4, or -7, which are known to enhance odontogenic differentiation. Since reactive oxygen species are also involved in the pulp toxicity of resin-based restorative materials, these results suggest that NAC may be a promising candidate for supplementation of dental restorative materials in order to enhance reparative dentine formation.

Background : This study, the fraction for testing the efficacy of the Astragalus extract was concentrated active ingredient. The concentrated fraction was applied to a cosmetic material that Astragalus testing results confirmed that the improved efficacy. Methods and Results : The fractions were performed using an n-butanol solvent for increasing the efficacy of the Astragalus extract, by using the material fractions collected to compare and ultimately an increase in whitening and wrinkle efficacy. The solvent to be used in the fractions was used for the n-butanol good dissolution to the effective substance(Astragaloside, Isoflavonoid). It increased approximately 6.5 times the sample extract and the n-butanol fraction of the Astragalus as a result Astragaloside 15 ppm, 97 ppm respectively analyzed by HPLC equipment, isoflavonoid content was confirmed by an increase in the content of the fractions increased 4.5 times to 280 ppm, 1,260 ppm. Tyrosinase inhibitory effect, respectively IC50 5.70 mg/mL, IC50 1.02 mg/mL to, Collagenase producing ability is IC50 4.88 mg/mL, IC50 0.93 mg/mL with n-butanol fraction was good whitening, anti-wrinkle efficacy than the extract. Sensory evaluation was conducted in the same amount of sample, using a purified Astragalus cosmetics received high marks. Stability evaluation(MTT assay) was checked for more than 100% cell viability at the concentration 2,000 ppm. Conclusion : n-butanol fraction of Astragalus was subjected to a component analysis and In vitro test, it was confirmed an increase active ingredient content. The results of sensory evaluation and stability evaluation, it was confirmed been made to improve qualities as a cosmetic materials.

Heat shock transcription factors(HSFs) are the major heat shock factors regulating the heat stress response. They participate in regulating the expression of heat shock proteins (HSPs), which are critical in the protection against stress damage and many other important biological processes. In this study, a genome-wide analysis was carried out to identify all HSFs soybean genes. Twenty six nonredundant HSF genes(GmHsf) were identified in the latest soybean genome sequence. Chromosomal location, protein domain and motif organization of GmHsfs were analyzed in soybean genome. The phylogenetic relationships, gene duplications and expression profiles of GmHsf genes were also presented in this study. According to their structural features, the predicted members were divided into the previously defined classes A–C, as described in Arabidopsis. Using RT-PCR, the expression patterns of 26 GmHsf genes were investigated under heat stress. The data revealed that these genes presented different expression levels in response to heat stress conditions. Real-time (q)RT-PCR was performed to investigate transcript levels of five GmHsfs in response to multiple abiotic stresses. Differential expression of five GmHsfs implies their role during abiotic stresses. Subcellular localization using GFP-fusion protein demonstrated that GmHsf12 and GmHsf34 were restricted to the nucleus and GmHsf28 was localized in the nucleus and cytoplasm in plant. The results provide a fundamental clue for understanding of the complexity of the soybean HSF gene family and cloning specific function genes in further studies and applications.

The plant-specific NAC (NAM, ATAF, and CUC)-domain proteins play important roles in plant development and stress responses. Comparative time-course expression analyses were carried out to analyze the expression levels of 62 soybean NAC genes during drought stress in order to search for the stress-inducible NAC genes. Ten GmSNAC (Glycine max stress-inducible NAC) genes having the significant differential expression in response to the drought stress and abscisic acid (ABA) hormone application were further investigated for their expression profiles with various stresses such as drought, high salinity, cold and with ABA treatments by the quantitative real-time PCR analyses. In this research, the full-length cDNAs of eight GmSNAC were isolated for the further studies. Eight GmSNAC proteins were tested for their transcription activation in the yeast assay system. Two GmSNAC proteins showed the very high transcriptional activities and the other two GmSNAC proteins displayed moderate levels of transactivation while the remaining four GmSNAC proteins lacked transactivation in yeast. Subcellular localization of eight GmSNAC proteins was analyzed via the green fluorescent protein-GmSNAC fusion protein in tobacco plant cell. Three GmSNAC proteins with the C-terminal transmembrane domain were localized to the nucleus and cytoplasmic fractions. The other five GmSNAC proteins were targeted to the nucleus. The function of GmSNAC49 gene was further investigated using the overexpression transgenic Arabidopsis. Germination rate in transgenic plants over-expressing GmSNAC49 was delayed in the media supplemented with mannitol or ABA compared with that of wild-type (WT) plants. The 35S:GmSNAC49 transgenic Arabidopsis displayed improved tolerance to drought stress compared to the WT. The results of this systematic analysis of the GmSNAC family responsive to abiotic stress will provide novel tools and resources for the development of improved drought tolerant transgenic soybean cultivars

Comparative time-course expression analyses were carried out to analyze the expression levels of 60 soybean WRKY genes during abiotic stress in order to search for the stress-inducible WRKY genes. Five GmWRKY(Glycine max WKRY) genes having the significant differential expression in response to the drought stress and abscisic acid(ABA) hormone application were further investigated for their expression profiles with various stresses such as drought, high salinity, cold and with ABA treatments by the quantitative real-time PCR analyses. In this research, the full-length cDNAs of five GmWRKY were isolated for the further studies. Five GmWRKY proteins were tested for their transcription activation in the yeast assay system. GmWRKY3 proteins showed the very high transcriptional activities and the other two GmWRKY proteins displayed moderate levels of transactivation while the remaining two GmWRKY proteins lacked transactivation in yeast. Subcellular localization of five GmWRKY proteins was analyzed via the green fluorescent protein-GmWRKY fusion protein in tobacco plant cell and all of GmWRKY proteins were targeted to the nucleus. In order to analyze the function of GmWRKY genes in plant, 35S:GmWRKY overexpression(OE) transgenic Arabidopsis were generated. Root growth and germination rates in transgenic OE plants were investigated in the media supplemented with mannitol, NaCl or ABA compared with that of wild-type(WT) plants. The 35S:GmWRKY42 transgenic Arabidopsis displayed reduced tolerance to drought stress compared to the WT. The results of this systematic analysis of the GmWRKY family responsive to abiotic stress will provide novel tools and resources for the development of improved drought tolerant transgenic soybean cultivars

Arabidopsis atDjC53 and atDjC32 gene DnaJ-like protein homologous to DnaJ-like protein was characterized for the functional analysis of DnaJ-like protein. It was shown that atDjC53 and atDjC32 RNA expression is induced by heat shock stress and atDjC53- and atDjC32-GFP was targeted to the nucleus of protoplasts. The atDjC53 and atDjC32 promoter (1 kb) was isolated and fused to the GUS reporter gene to investigate gene regulation of atDjC53 and atDjC32 specific to heat shock stress or to developmental organ in the transgenic lines. RNAi and overexpression construct was employed to generate atDjC53 and atDjC32 knock-out plants for the study of their function. Molecular function of atDjC53 and atDjC32 is discussed in relation to heat shock and also developmental stages in Arabidopsis.

Heat shock transcription factors (HSFs) are the major heat shock factors regulating the heat stress response. They participate in regulating the expression of heat shock proteins (HSPs), which are critical in the protection against stress damage and many other important biological processes. In this study, a genome-wide analysis was carried out to identify all HSFs soybean genes. Twenty six nonredundant HSF genes (GmHsf) were identified in the latest soybean genome sequence. Chromosomal location, protein domain and motif organization of GmHsfs were analyzed in soybean genome. The phylogenetic relationships, gene duplications and expression profiles of GmHsf genes were also presented in this study. According to their structural features, the predicted members were divided into the previously defined classes A–C, as described in Arabidopsis. Using RT-PCR, the expression patterns of 26 GmHsf genes were investigated under heat stress. The data revealed that these genes presented different expression levels in response to heat stress conditions. Real-time (q)RT-PCR was performed to investigate transcript levels of five GmHsfs in response to multiple abiotic stresses. Differential expression of five GmHsfs implies their role during abiotic stresses. Subcellular localization using GFP-fusion protein demonstrated that GmHsf12 and GmHsf34 were restricted to the nucleus and GmHsf28 was localized in the nucleus and cytoplasm in plant. The results provide a fundamental clue for understanding of the complexity of the soybean HSF gene family and cloning specific function genes in further studies and applications.

Salinity stress severely affects plant growth and development causing crop loss worldwide. Suaeda asparagoides is a salt-marsh euhalophyte widely distributed in southwestern foreshore of Korea. To isolate salt tolerance genes from S. asparagoides, we constructed a cDNA library from leaf tissues of S. asparagoides that was treated with 200 mM NaCl. A total of 1,056 clones were randomly selected for EST sequencing, and 932 of them produced readable sequence. By sequence analysis, we identified 538 unigenes and registered each in National Center for Biotechnology Information. The 80 salt stress related genes were selected to study their differential expression. Reverse Transcriptase-PCR and Northern blot analysis revealed that 23 genes were differentially expressed under the high salinity stress conditions in S. asparagoides. They are functionally diverse including transport, signal transduction, transcription factor, metabolism and stress associated protein, and unknown function. Among them dehydrin (SaDhn) and RNA binding protein (SaRBP1) were examined for their abiotic stress tolerance in yeast (Saccharomyces cerevisiae). Yeast overexpressing SaDhn and SaRBP1 showed enhanced tolerance to osmotic, freezing and heat shock stresses. This study provides the evidence that SaRBP1 and SaDhn from S.asparagoides exert abiotic stress tolerance in yeast. Information of salt stress related genes from S. asparagoides will contribute for the accumulating genetic resources to improve osmotic tolerance in plants.

The ubiquitin conjugating enzyme E2 (UBC E2) mediates selective ubiquitination, acting with E1 and E3 enzymes to designate specific proteins for subsequent degradation. In the present study, we characterized the function of the mung bean VrUBC1 gene (Vigna radiata UBC 1). RNA gel-blot analysis showed that VrUBC1 mRNA expression was induced by either dehydration, high salinity or by the exogenous abscisic acid (ABA), but not by low temperature or wounding. Biochemical studies of VrUBC1 recombinant protein and complementation of yeast ubc4/5 by VrUBC1 revealed that VrUBC1 encodes a functional UBC E2. To understand the function of this gene in development and plant responses to osmotic stresses, we overexpressed VrUBC1 in Arabidopsis (Arabidopsis thaliana). The VrUBC1-overexpressing plants displayed highly sensitive responses to ABA and osmotic stress during germination, enhanced ABA- or salt-induced stomatal closing, and increased drought stress tolerance. The expression levels of a number of key ABA signaling genes were increased in VrUBC1-overexpressing plants compared to the wild-type plants. Yeast two-hybrid and bimolecular fluorescence complementation demonstrated that VrUBC1 interacts with AtVBP1 (A. thaliana VrUBC1 Binding Partner 1), a C3HC4-type RING E3 ligase. Overall, these results demonstrate that VrUBC1 plays a positive role in osmotic stress tolerance through transcriptional regulation of ABA-related genes and possibly through interaction with a novel RING E3 ligase.