Hypocreales entomopathogenic Beauveria bassiana and Metarhizium anisopliae are well-known biological control agents worldwide and have high potential in industrialization. However their thermo-susceptibility limits long-term storage under high temperature conditions and high insecticidal activity after application to target pests. Herein we isolated highly virulent isolates, B. bassiana JEF006 and JEF007 and M. anisopliae JEF003 and 004, and produced in three grains, as substrates for solid cultures, followed by thermotolerance assays to compare the potential of the three substrates. The JEF isolates were exposed to dry and wet heat at 50°C and overall conidia were more stable under dry heat condition rather than wet heat. Of the three grains, Italian millet was superior to the other grains in the production of thermotolerant conidia. Additionally Italian millet did not severely aggregated, which enabled air to penetrate into the substrate well compared to the sorghum and millet. JEF isolates were more thermotolerant when they were kept in oil conditions. The conidia of JEF 007 and 003 wild type and AtMT-based generating random mutants were subjected to SDS-PAGE. A significant relationship between conidial thermotoelrance and detected proteins was observed. This work suggests that Italian millet can be used as an effective substrate to produce more thermotolerant conidia, thus maintaining viability for long times under unfavorable environment and biological activity against target pests.

Entomopathogenic Beauveria bassiana and Metarhizium anisopliae are well-known biological control agents worldwide and have high potential in industrialization. However their thermo-susceptibility limits long-term storage under high temperature conditions and high insecticidal activity after application to target pests. Herein we isolated highly virulent isolates, B. bassiana JEF006 and JEF007 and M. anisopliae JEF003 and JEF004, and produced in three grains, such as sorghum, millet and Italian millet as substrates for solid cultures, followed by thermotolerance assays to compare the potential of the three substrates for thermotolerance. The JEF isolates were exposed to dry and wet heat at 50°C and overall conidia were more stable under dry heat condition rather than wet heat. Of the three grains, Italian millet was superior to the other grains in the production of thermotolerant conidia. Additionally Italian millet did not severely aggregated, which enabled air to penetrate into the substrate well compared to the sorghum and millet. JEF isolates were more thermotolerant when they were kept in oil conditions as carriers of an oil-based formulation. This work suggests that Italian millet can be used as an effective substrate to produce more thermotolerant conidia, thus maintaining viability for long times under unfavorable environment and biological activity against target pests.

The effects of seed soaking treatment with the solutions of plant growth regulators IAA, GA3 and BAP on seed germination and shoot and bulb growth of Allium victorialis var. platyphyllum (Korean wild garlic) were determined. A significant variation in the seed germination rate was recorded at all treatments for various soaking periods. Maximum seed germination was obtained when seeds were soaked in IAA or GA3 solution at 200 mg L-1. The MAP treated seeds started to germinate after 3 months. Among treatments, IAA was found to be most effective in improving seed germination, but further seedling growth was not correlated to the soaking time. Seed soaking in IAA or GA3 solution enhanced further growth of seedlings compared with water control treatment. Shoot and bulb growth was highest in GA3 treatments.

Germinated rough rice (GRR) has many healthy effects such as antioxidant activity, inhibition of chronic inflammatory disease and prevention of vascular disease. In this study, germinated Korean rice sample which was planted in 2008 from Korean Rural Development Administration was used. The purpose of this study was to recognize the effect of the ethanol extract of GRR on the ischemic damage of skeletal muscle. Right femoral artery ligation was performed to induce ischemic condition in mice and the mixed basal diet with the ethanol extract of GRR at various levels (100, 200, and 400 mg/kg diet) was given for 2 weeks. After the mice were sacrificed, the hind limb muscle samples of the mice were fixed, sectioned, stained with Haematoxylin-Eosin (H&E), Masson’s trichrome, and performed for Immunohistochemistry (IHC) for caspase-3. In histopathological findings of H&E and Masson’s trichrome stain, the treatment of GRR at the level of 400 mg/kg diminished fibrosis and necrosis in the muscle compared to the control. The expression of caspase-3 protein was also decreased by the treatment of GRR at the level of 400 mg/kg compared to the control. These results suggest that the ethanol extract of GRR may have a protective effect against muscle damage during the ischemia in skeletal muscle of hindlimb.

4-Nonylphenol (NP) is a surfactant that is a well-known and widespread estrogenic endocrine disrupting chemical (EDC). Although it has been known that the affinity of NP to ERs is low, it has been suggested that low-dose NP has toxicity. In the present study, the endocrine disrupting effects on reproduction, and the weight of gonads, epididymis, and uterus were evaluated with the chronic lower-dose NP exposing. This study was designed by following the OECD test guideline 443 and subjected to a complete necropsy. In male, NP had an effect on the weight of the testis and epididymis in both F0 and F1. In females, NP decreased the weight of ovary and uterus in F0 but not in pre-pubertal F1 pubs. Fertility of male and female in F0 or F1 was no related with NP administration. The number of caudal-epididymal sperm by body weight (BW) was not different between groups in both F0 and F1. Besides, the difference of the sperm number between generations was not detected. The number of ovulated oocytes was similar between groups in F0, but significantly decreased in NP 50 group of F1. The litter size and sex ratios of offspring in F1 and F2 were not different. The accumulated mating rate and gestation period were not affected by the NP administration. Those results shows that chronic lower-dose NP administration has an effect of endocrine disruptor on the weight of gonads and epididymis of F0 and F1 but not in reproduction. Based on the results, it is suggested that chronic lower-dose NP exposing causes endocrine disruption in the weight of gonad and epididymis but not in the reproductive ability of next generations.

Background : The geographical origin of Panax ginseng Meyer, a valuable medicinal plant, is important to both ginseng producers and consumers in the context of economic profit and human health benefits. We therefore aimed to discriminate between the cultivation regions of ginseng using the stable isotope ratios of C, N, O, and S, which are abundant bio-elements in living organisms. Methods and Results : The C, N, O, and S stable isotope ratios were measured by isotope ratio mass spectrometer, and then these isotope ratios profiling was statistically analyzed with chemometrics. The various isotope ratios found in Panax ginseng roots were significantly influenced by region, cultivar, and the interactions between these two factors (P ≤ 0.0002). In particular, δ18O was lower in ginseng roots grown at high altitudes (r = −0.47), while δ34S was higher in ginseng roots grown close to coastal areas (r = −0.48). Chemometric results provided discrimination between the majorities of different cultivation regions. Conclusion : Our case study extends the understanding about the variation of C, N, O, and S stable isotope ratios in ginseng root depending on cultivation region. Hence, the analysis of stable isotope ratios is a suitable tool for discrimination between the regional origins of ginseng samples from Korea, with potential application to other countries.

Background : Corrosion is one of the most devastating problems faced by most industries. Mild steel has played a vital role in various fields due to the excellent mechanical properties of mild steel such as low density, high strength-to-weight ratios, good environmental stability, high thermal conductivity, and corrosion resistance. Methods and Results : The total phenolic contents (TPC) and total flavonoid contents (TFC) of the methanolic extract of C. grandiflora and R. verniciflua leaf have been examined, and its corrosion inhibition performance was investigated by weight loss and electrochemical impedance spectroscopy (EIS) and polarization measurements. The surface morphology of mild steel was analyzed by scanning electron microscopy (SEM) equipped with energy dispersive X-ray spectroscopy (EDX) and by atomic force microscopy (AFM). The percentage composition of polyphenolic compounds was found to be higher in C. grandiflora and R. verniciflua plant extracts, and it was proved to be a superior, eco-friendly, and anti-corrosive inhibitor for mild steel in 1M of H2SO4. The Tafel polarization studies indicate that the plant extract is a mixed-type inhibitor. Scanning electron microscopy/energy -dispersive X-ray spectroscopy (SEM-EDS), and atomic force microscopy (AFM) studies confirmed the formation of a protective film on the metal surface. The corrosion inhibition of the C. grandiflora and R. verniciflua plant extracts was characterized by Fourier transform infrared (FT-IR), UV-visible spectra, and wide-angle X-ray diffraction (XRD) studies; these show the strong interaction between the metal surface and the inhibitor. Conclusion : The methanolic extract was prepared the two different plants like C. grandiflora, and R. verniciflua was studied the corrosion inhibition on the mild steel specimen in acidic medium through various methods involving weight loss measurements, EIS, and potentiodynamic polarization. The results shows that the C. grandiflora, and R. verniciflua plant extracts illustrate an effective corrosion inhibitor for mild steel with good anticorrosion properties in acidic environmen

Background : The natural stable isotope ratio of common bio-elements like carbon (C), nitrogen (N), oxygen (O), or sulfur (S) varies with diverse isotope fractionation processes in nature. Therefore, measuring the variation of these stable isotope ratios in ginseng roots can be a feasible tool to discriminate the geographical origins of ginseng in Korea. Methods and Results : The 3-year-old six Korean ginseng cultivars were cultivated at the five regions in Korea, and then used for measuring the stable isotope ratios of C, N, O, and S by isotope ratio mass spectrometry (IRMS). The mean C, N, O, and S stable isotope ratio values in the ginseng roots significantly differed according to the cultivation regions (p < 0.05). However, these isotope ratios in ginseng roots had relatively weak discriminative power against to the ginseng cultivars at each cultivation region. The interaction of the cultivation region and ginseng cultivar type also significantly affected to the C, N, O, and S stable isotope ratio in ginseng roots (p < 0.0001). The two-dimensional plots associated with the N stable isotope ratio can effectively separate the ginseng roots in Jinan compared to those in the other regions. The partial least squares-discriminant analysis showed more significant separation between ginseng geographical origins compared to the principal component analysis. Conclusion : Our findings improve our understanding of how the isotope composition of ginseng roots varies with respect to cultivation regions and cultivars, and suggest that the analysis of the stable isotope ratios combined with chemometrics can be used as a feasible tool to discriminate geographical origin of ginseng in Korea.

The flower buds of Sophora japonica L (SF), as a well-known traditional Chinese medicinal herb, have been used to treat bleeding-related disorders such as hematochezia, hemorrhoidal bleeding, dysfunctional uterine bleeding, and diarrhea. However, no specific anti-cancer effect and its molecular mechanism of SF have been described. Thus, we performed in vitro study to investigate if treatment of SF affects activating transcription factor 3 (ATF3) expression and ATF3-mediated apoptosis in human colorectal cancer cells. The effects of SF on cell viability and apoptosis were measured by MTT assay and Western blot analysis against cleaved poly (ADP-ribose) polymerase (PARP). ATF3 activation induced by SF was evaluated using Western blot analysis, RT-PCR and ATF3 promoter assay. SF treatment caused decrease of cell viability and increase of apoptosis in a dose-dependent manner in HCT116 and SW480 cells. Exposure of SF activated the levels of ATF3 protein and mRNA via transcriptional regulation in HCT116 and SW480 cells. Inhibition of extracellular signal-regulated kinases (ERK) 1/2 by PD98059 and p38 by SB203580 attenuated SF-induced ATF3 expression and transcriptional activation. Ectopic ATF3 overexpression accelerated SF-induced cleavage of PARP. These findings suggest that SF-mediated apoptosis may be the result of ATF3 expression through ERK1/2 and p38-mediated transcriptional activation.

UGT72E3/2 gene encodes UDP-glycosyltransferase shown to glucosylate several phenylpropanoids such as syringin and coniferin. Syringin has effect of anti-stress and anti-fatigue. Korean soybean variety Kwangan was transformed with UGT72E3/2 gene. This gene was transformed into Kwangan using highly efficient soybean transformation system. This study used two promoters, beta-conglycinin promoter for seed-specific expression and 35s promoter for total expression. Transgenic plants were confirmed for gene introduction and their expression using PCR and RT-PCR. The analysis of syringin in transgenic plants was performed using HPLC. Currently, the confirmation of stable gene introduction with UGT72E3/2 gene is also performing by Southern blot analysis.

Soybean mosaic virus (SMV), a member of Potyviridae family, is one of the most typical viral diseases and results in yield and quality loss of cultivated soybean. Due to the depletion of genetic resources for resistance breeding, a trial of genetic transformation to improve disease resistance has been performed by introducing SMV-CP and HC-Pro gene by RNA interference (RNAi) method via Agrobacterium-mediated transformation. Transgenic plants were infected with SMV strain G5 and investigated the viral response. As a result, two lines (3 and 4) of SMV-CP(RNAi) transgenic plants and three lines (2, 5 and 6) of HC-Pro(RNAi) transgenic plants showed viral resistance. In genomic Southern blot analysis, most of lines contained at least one T-DNA insertion in both SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. Subsequent investigation confirmed that no viral CP and HC-Pro gene expression was detected in two SMV-resistant lines of SMV-CP(RNAi) and three lines of HC-Pro(RNAi) transgenic plants, respectively. On the other hand, non-transgenic plants and other lines showed viral RNA expression. Viral symptoms affected seed morphology, and clean seeds were harvested from SMV-resistant line of SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. In addition, strong viral gene expression was detected from seeds of SMV-susceptible non-transgenic plants and SMV-susceptible transgenic lines. When compared the viral resistance between SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants, soybean transgenic plants with the HC-Pro gene using RNAi strategy showed much stronger and higher frequency of viral resistance.

Bacillus thuringiensis(Bt) crystal protein (Cry1Ac) genes encode insecticidal δ-endotoxins that are widely used for the development of insect-resistant crops. Common soybean is a crop of economic and nutritious importance in many parts of the world. Korea soybean variety Kwangan was transformed with Bacillus thuringiensis(Bt) crystal protein genes. We transformed three difference Cry1Ac (Cry1Ac and two modified Cry1Ac) genes into Kwangan using highly efficient soybean transformation system. Transgenic plants with Bt crystal protein genes were confirmed for gene introduction and their expression using PCR, real-time PCR, and RT-PCR. We generated 30 independent lines of transgenic soybean plants. Analysis of the flanking sequences isolated by Inverse PCR revealed complex T-DNA insertion patterns and preferential integration of T-DNA into the intergenic spacer region of the soybean genome. We found 5 different intergenic transgenic soybean lines of soybean genome. Currently, the confirmation of stable gene introduction with Bt genes is also performing by southern blot analysis, physiology test, and agronomic characters are investigating.

Twenty two common millet (Panicum miliaceum L.) varieties collected from Korea, China and Russia were investigated for their phylogenetic relationship using 5S ribosomal DNA sequences with a hope to provide the basic information on their exact origin. Sequences of 5S rDNA were isolated by PCR. The primers, 5s-rRNA1 and 5s-rRNA2, were designed to isolate the complete NTS. Genomic DNA amplification produced two fragments with different length, 900 bp and 400 bp fragments, confirming the presence of two types of 5S rDNA repeats that differed from each other in the length of the NTS region. Amplified DNAs of 400 bp fragment were subcloned and used for further investigation. The obtained NTS sequences ranged from 200 to 300 bp and homology of sequences among plant materials was much higher than long repeat. CLUSTALW multiple aligment of 5S rDNA sequences from 22 different common millets revealed the clear difference by their origin. And critically different areas with insert or deletion were also confirmed. Those sequence difference seems to be used for discrimination of cultivars from different origin and use as molecular markers for origin identification. In phylogenic tree construction, the clear classification was shown where the genotypes from China and Russia is positioned together and stay away from domestic genotypes.

Rye has important genes for biotic and abiotic stress resistance. Introduction of these genes to wheat by breeding wheat-rye translocation have been intensively used in wheat breeding program. Rye chromatin 1RS and/or 2RL show superior performance in unfavorable environments. In order to develop high yielding wheat, we applied various molecular breeding strategies. To develop EST-derived 1RS specific markers, we used comparative genomics with public sequence databases of Poaceae family. Putative rye chromatin specific sequences were used to design 1RS specific markers. To identify genes related to water deficiency, cDNA AFLP analysis was used in PEG treated seedlings of 1RS RILs. For functional analysis of identified genes and markers, we used Brachypodium distachyon, as a new model plant of temperate grasses. B. distachyon were recently applied for transformation and we constructed Agrobacterium-mediated transformation system. Integration of those strategies and conventional breeding method would enhance the usefulness of rye chromatins for wheat improvement.

Wild rice might have previously unidentified genes important for disease resistance and stress tolerance in response to biotic and abiotic stresses. A set of subtractive library was constructed both from leaves of wild rice plants, Oryza grandiglumis (CCDD, 2n=48), treated with fungal elicitor and from wounded leaves. A partial fragment that was homologous to PR10 genes from other plant species was identified via suppression subtractive hybridization and cDNA macroarray. The obtained full-length cDNA sequence (OgPR10) contains an open reading frame of 480 bp nucleotide, encoding 160 amino acids with a predicted molecular mass of 16.944 kDa and an isoelectric point (pI) of 4.91. The multiple alignment analyses showed the higher sequence homology of OgPR10 with PR10 genes identified in rice plants at amino acid level. The OgPR10 mRNA was not expressed by treatment with wounding, jasmonic acid, and salicylic acid, but markedly expressed in leaves treated with protein phosphatase inhibitors cantharidin and endothall, and yeast extract. In addition, the expression of OgPR10 mRNA was induced within 72 h after treatment with probenazole, one of well-known chemical elicitors, and reached the highest level at 144 h. Heterologous expression of OgPR10 caused growth inhibition and seedling lethality in E. coli and Arabidopsis, respectively. Chemically induced OgPR10 expression with glucocorticoid-mediated transcriptional induction system further reconfirmed its lethality on Arabidopsis seedling. In addition, OgPR10-expressing rice plants, Oryzae sativar were resistant against the infection of rice blast fungus, Magnaporthe grisea. These results indicate that OgPR10 is involved in probenazole- and microbe associated molecular patterns-mediated disease resistance responses in plants and is a potential gene for developing disease resistance crop plants.