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Panax ginseng is a major medicinal crop with pharmaceutical efficacy derived from ginsenoside metabolites. Despite its genome information, the inefficiency of ginseng transformation hinders the study of the molecular mechanism of ginseng plant metabolism. Thus, this protocol aimed to develop an easy and efficient system for ginseng transformation. We established a transformation system using ginseng callus cultured in a liquid medium, which has a higher ratio compared with cotyledon explant. In addition, Agrobacterium tumefaciens has been used for plant transformation. Compared with the LBA4404 strain, C58C1 was inappropriate for ginseng transformation using ginseng callus. We induced and maintained calli in liquid medium and cut them into small pieces before infection. After infection, we selected calli that survived from the antibiotic medium until identification of newly growing cells. In β-glucuronidase (GUS) gene assay, the expression of the GUS gene was observed in cells that were newly generated from explants. We treated calli with 0.05 M of MgSO4 before infection. After MgSO4 pre-treatment, the transformation efficiency of growing cells around infected callus was increased. Moreover, we constructed and introduced a visible reporter RUBY system to easily identify transformed cells. Using this system, we could identify the cells by a red color with naked eyes. Based on our transformation protocol, the success rate has increased to 77.27% in surviving lines during selection culture. This stable ginseng transformation could facilitate the overexpression and knockout of ginseng lines for functional or synthetic biological studies.

Oxidative stress has been demonstrated to play a pivotal role in the pathological processes of many neurodegenerative diseases. In the present study, we demonstrated that Chrysanthemum boreale Makino extract (CBME) suppresses oxidative stress-induced neurotoxicity in human neuroblastoma SH-SY5Y cells and elucidated the underlying molecular mechanism. Our observations revealed that CBME effectively protected neuronal cells against H2O2-induced cell death by preventing caspase-3 activation, Bax upregulation, Bcl-2 downregulation, activation of three mitogen-activated protein kinases (MAPKs), cAMP response element-binding protein (CREB) and NF-κB phosphorylation, and iNOS induction. These results provide evidence that CBME has remarkable neuroprotective properties in SH-SY5Y cells against oxidative damage, suggesting that the complementary or even alternative role of CBME in preventing and treating neurodegenerative diseases is worth further studies.

Photodynamic therapy (PDT) is a promising cancer treatment with fewer side effects, and it eliminates tumors in target tissues with reactive oxygen species produced by photosensitizers (PS) and light. In this study, we isolated methyl pheophorbide A, which induces photodynamic cell death in the U937 and SK-HEP-1 cells, from perilla leaves. Its potential as a material for the development of a new PS was also evaluated. The methyl pheophorbide A is a dark green porphyrin compound isolated from methanol extract of perilla leaves. Apoptosis occurred was methyl pheophorbide A treated and irradiated with light, and there was no significant change where light not treated. In both U937 and SK-HEP-1 cells, apoptotic body, vesicle formation, and DNA ladder were confirmed in the light-irradiated. The caspase-3/7 activity an important factor in apoptosis, was 101.50 ± 14.24% when treated with 0.25 μg/ml methyl pheophorbide A in U937, and 91.32 ± 16.23% when treated with 1.00 μg/ml in SK-HEP-1 cells. The apoptotic phenomenon appeared more strongly when the methyl pheophorbide A concentration was increased with irradiating light. This study demonstrates the isolation, identification, and phototoxic activity of methyl pheophorbide A in perilla leaves. We expect that this study will be useful in the search for PS candidates using natural products.

Cytochrome P450 (CYP) catalyzes a wide variety of monooxygenation reactions in plant primary and secondary metabolisms. Land plants contain CYP703, belonging to the CYP71 clan, which catalyzes the biochemical pathway of fatty acid hydroxylation, especially in male reproductive tissues. Korean/Asian ginseng (Panax ginseng Meyer) has been regarded as one of important medicinal plant for a long time, however the molecular mechanism is less known on its development. In this study, we identified and characterized a CYP703A gene in P. ginseng (PgCYP703A4), regarding reproductive development. PgCYP703A4 shared a high-sequence identity (81–83%) with predicted amino acid as CYP703 in Dancus carota, Pistacia vera, and Camellia sinensis as well as 76% of amino acid sequence identity with reported CYP703 in Arabidopsis thaliana and 75% with Oryza sativa. Amino acid alignment and phylogenetic comparison of P. ginseng with higher plants and known A. thaliana members clearly distinguish the CYP703 members, each containing the AATDTS oxygen binding motif and PERH as a clade signature. The expression of PgCYP704B1 was only detected in P. ginseng flower buds, particularly in meiotic cells and the tapetum layer of developing anther, indicating the conserved role on male reproduction with At- and Os- CYP703. To acquire the clue of function, we transformed the PgCYP703A4 in A. thaliana. Independent overexpressing lines (PgCYP703A4ox) increased silique size and seed number, and altered the contents of fatty acids composition of cutin monomer in the siliques. Our results indicate that PgCYP703A4 is involved in fatty acid hydroxylation which affects cutin production and fruit size.

Cytochrome P450 (CYP) catalyzes a wide variety of monooxygenation reactions in plant primary and secondary metabolisms. Land plants contain CYP703, belonging to the CYP71 clan, which catalyzes the biochemical pathway of fatty acid hydroxylation, especially in male reproductive tissues. Korean/Asian ginseng (

Type 2 diabetes mellitus (T2DM) and osteoarthritis (OA) are common chronic diseases that frequently co-exist. The link between OA and T2DM is attributed to common risk factors, including age and obesity. Several reports suggest that hyperglycemia and accumulated advanced glycosylation end-products might regulate cartilage homeostasis and contribute to the development and progression of OA. Metformin is used widely as the first-line treatment for T2DM. The drug acts by regulating glucose levels and improving insulin sensitivity. The anti-diabetic effects of metformin are mediated mainly via activation of adenosine monophosphate (AMP)-activated protein kinase (AMPK), which is an energy sensing enzyme activated directly by an increase in the AMP/ATP ratio under conditions of metabolic stress. Dysregulation of AMPK is strongly associated with development of T2DM and metabolic syndrome. In this review, we discuss common risk factors, the association between OA and T2DM, and the role of AMPK. We also address the adaptive use of metformin, a known AMPK activator, as a new drug for treatment of patients with OA and T2DM.

Lime bordeaux mixture (LBM) and lime sulfur mixture (LSM) are representative environmental friendly organic materials for prevention of insect pests in South Korea. Recently, those have been widely used as an alternative for chemical pesticides in eco-friendly farms. However, South Korea has not established even recommendation of LBM and LSM considering the stability of heavy metals in soil. The aim of this study was to evaluate the accumulation of hazardous heavy metals in soil and plant with long-term application of LBM and LSM. Firstly, we investigated the amount of LBM and LSM used per year in several eco-friendly farms to determine a standard application rate of both materials. The pepper plant was grown on the pot in greenhouse for 14 weeks. Both materials were applied at 0, 1, 3, and 9 times of standard application rates (2.56 and 1.28 L ha-1 of LBM and LSM per year, respectively). Dry matter yield of pepper and heavy metals (As, Cd, Cu, Hg, Ni, Pb, and Zn) concentration in soil and pepper plant were measured after 14 weeks. Yield of pepper plant did not significantly chang with up to application rate of 1 times, thereafter it markedly decreased with more than 3 times. With increasing LBM and LSM application, the concentration of Cu and Zn in soil significantly increased. Especially, Zn concentration in pepper significantly increased with increasing application rates of both materials. This might resulted in significant decrease in dry matter yield of pepper. The concentrations of those heavy metals in soil did not exceed safety levels (150 mg kg-1 for Cu and 300 mg kg-1 for Zn) established by the Korean Soil Environmental Conservation Act as well as concentration of heavy metals in pepper plant by Korean Ministry of Food and Drug Safety. However, particular attention should be paid for heavy metal safety and crop productivity when using LBM and LSM in the organic farm.

Cordyceps is a traditional Chinese medicinal herb well-known in China, Korea and Japan since B.C. 2,000. The original entomopathogenic fungus, Cordyceps sinensis belonging to the genus Cordyceps could not be found inside Korean peninsula due to the absence of the host insect for the corresponding entomogenous fungus. The development of artificial production methods of Korean type Cordyceps using the silkworm Bombyx mori as in vivo culture medium for the the entomopathogenic fungus Paecilomyces tenuipes is the first, and wonderful occasion in the research history of insect industry of this global world. The aim of this article is to review the historical research background, mass-production methods, and pharmacological effects of the silkworm-dongchunghacho (Paecilomyces tenuipes) which is a newly developed Korean medicinal insect-borne mushroom, and another non-insect-borne medicinal mushroom (Cordyceps militaris and Cordyceps pruinosa). Their biological actions include anti-tumor, immunostimulating, anti-fatigue, anti-stress, anti-oxidant, anti-aging, anti-diabetic, anti-inflammatory, anti-thrombosis, hypolipidaemic and insecticidal effects. The bioactive principles are protein-bound polysaccharides (hexose, hexosamin), cordycepin, D-manitol, acidic polysaccharide etc. Protein-bound polysaccharides and n-butanol fractions were demonstrated to show a significant anti-tumor activities but did not show a cytotoxicities. D-mannitol exhibited a significant prolongation of the life span in tumor bearing mice. Ergosterol did not show an efficient anti-tumor activity, but showed a significant phagocytosis enhancing activity. Anti-tumor activity of silkworm-dongchunghacho might be attributed to immuno-stimulating activities rather than cytotoxic effects [164]. Also this review comprises the breeding of Dongchunghacho varieties, optimization of culture conditions, improvement of learning and memory by Dongchunghacho, application of them as foods and chemical constituents.

Various types of organic materials could affect differently immobilization of cadmium (Cd) and its uptake by plant grown in soil. Therefore, this study was conducted to evaluate effect of different organic materials in reducing Cd phytoextractability in contaminated arable soil. To do this, rice straw and composted manure were selected as organic materials and applied at the rate of 0, 15, 30, and 45 Mg ha-1 in Cd contaminated arable soil with 6.5 mg kg -1 of total Cd. Radish (Raphanus sativa L.) was seeded and grown for 50 days to evaluate Cd phytoavailability with different organic materials. Composted manure was more effective to decrease 1 M NH₄OAc extractable Cd concentration and increase pH of soil than rice straw. One M NH₄OAc extractable Cd concentration significantly decreased with increasing application rate of composted manure. Tendency of Cd uptake by radish plant with application of different organic materials was similar to that of 1 M NH₄OAc extractable Cd concentration and soil pH. Changes of soil pH with application of straw and composted might be one of factors to determine extractability and phytoavailability of Cd in this study. Radish yield significantly increased with up to 45 Mg ha -1 of composted manure application but did not with straw application. In the view point of Cd phytoextractability and plant productivity, it is recommended to apply composted manure rather than straw in Cd contaminated arable soil.

Adipogenesis is one of the cellular processes and a highly controlled program. Nowadays, inhibition of adipogenesis has received attention as an effective way to regulate obesity. In the current study, we investigated the inhibition effect of a chloroform extract of Pleurotus eryngii var. ferulae 'Beesan No. 2' (CEBT) on adipogenesis in 3T3-L1 murine preadipocytes. Pleurotus eryngii var. ferulae is one of many varieties of King oyster mushroom and has been reported to have various biological activities, including antitumor and anti-inflammation effects. Biological activities of 'Beesan No. 2', a new cultivar of Pleurotus eryngii var. ferulae, have not yet been reported. In this study, we found that CEBT suppressed adipogenesis in 3T3-L1 cells through inhibition of key adipogenic transcription factors, such as peroxisome proliferator-activated receptor γ and CCAAT/enhancer binding protein α. Additionally, CEBT reduced the expression of the IRS/PI3K/Akt signaling pathway and its downstream factors, including mammalian target of rapamycin and p70S6 kinase, which stimulate adipogenesis. Furthermore, β-catenin, a suppressor of adipogenesis, was increased in CEBT-treated cells. These results indicate that Pleurotus eryngii var. ferulae 'Beesan No. 2' effectively inhibited adipogenesis, so this mushroom has potential as an anti-obesity food and drug.

The contents of functional components in the organically agricultural perilla leaves were compared with the perilla leaves grown in conventional condition. Perilla leaves used in the experiment were purchased or harvested from each three organic farm houses and conventional farm houses in Miryang city. The analyzed components included total phenol, total flavonoid, vitamin C, vitamin E, β-carotene, GABA, caffeic acid, rosmarinic acid, Total N, Zn, Fe, Ca, Mg, Na, K, and P. Organically grown perilla leaves had vitamin E 49% greater than the conventional perilla leaves, and 21% for TN, 29% for Ca, and 73% for Mg, while conventionally grown perilla leaves contained K 16% higher than organic ones. Other components were not showed the differences.

BACKGROUND: Since bottom ash (BA) contains considerable amounts of CaO and MgO, it could be a useful amendment to increase soil pH and to immobilize cadmium (Cd). This study was conducted to evaluate effect of BA application in reducing Cd phytoavailability.METHODS AND RESULTS: Bottom ash was applied at the rate of 0, 20, 40, and 80 Mg/ha to Cd contaminated soil, and then lettuce was cultivated under field condition. soil pH and net negative charge increased slightly with increasing BA application; however, there was no statistical difference among the rates. Water soluble, exchangeable+acidic, reducible, and oxidizable fraction of Cd decreased with increasing bottom ash application rate, whereas residual fraction of Cd increased with increasing bottom ash application rate. Lettuce yield increased with rate of bottom ash up to 40 kg/ha. Visual evidences of cadmium toxicity and growth inhibition were not found during lettuce cultivation.CONCLUSION: Bottom ash was effective to reduce phytoextractability of Cd and to increase lettuce yield. Conclusively, BA could be a good soil amendment to reduce Cd phytoavailability in contaminated arable soil.

BACKGROUND: Petroleum-contaminated soil from leaking above- and underground storage tanks and spillage during transport of petroleum products is widespread environmental problem in recent years. Application of compost may be the most promising, cost-effective, and eco-friendly technology for soil bioremediation because of its advantages over physical and chemical technology. The objective of this study was to evaluate effect of compost application on degradation of total petroleum hydrocarbon (TPH) in petroleum hydrocarbon-contaminated soil. METHOD AND RESULTS: An arable soil was artificially contaminated by diesel, and compost was applied at the different rate of 0, 10, 30, and 50 Mg/ha. Concentration of TPH in the soil decreased as application rate of compost increased. Degradation efficiency was highest at compost 30 Mg/ha; however, it slightly decreased with compost 50 Mg/ha. Kinetic modeling was performed to estimate the rates of chemical reaction. The correlation coefficient (R2) values for the linear plots using the second-order model were higher than those using the first-oder model. Compost 30 and 50 Mg/ha had the fastest TPH degradation rate in the second-order model. Change of microbial population in soil with compost application was similar to that of TPH. Microbial population in the soil increased as application rate of compost increased. Increasing microbial population in the contaminated soil corresponded to decreased in TPH concentration. CONCLUSION: Conclusively, compost application for soil bioremediation could be an effective response to petroleum hydrocarbon-contaminated soil. The increase in microbial population with compost suggested that compost application at an optimum rate might enhance degradation of TPH in soil.

The objectives of this study were to determine (1) the phosphorus (P) level required to induce cadmium (Cd) precipitation in a contaminated arable soil with low concentrations of Cd and (2) the primary mechanism of Cd immobilization at different P levels. Phosphorus was added at levels of 0 800, 1600, and 16,000 mg P kg(-1) to a soil containing 5.57 mg Cd kg(-1). The concentration of 1 M NH4OAc extractable Cd decreased significantly with P levels up to 1600 mg kg(-1) due to an increase in soil pH and negative charge of soil (p

In this study, we tried to separate the photosensitizer that induces apoptosis of leukemia cells (U937) from perilla leaves. Perilla leaves (Perilla frutescens Britt var. japonica Hara) are a popular vegetable in Korea, being rich in vitamins (A and E), GABA, and minerals. Dried perilla leaves were extracted with methanol to separate the ph otosensitizer by various chromatographic techniques. The structure of the isolated compound (PL9443) was identified by 1D-NMR, 2D- NMR, and FAB-mass spectroscopy. Absorbance of the UV-Vis spectrum was highest at 410 nm and was confirmed by the 330, 410, and 668 nm. PL9443 compound was determined to be pheophorbide, an e thyl ester having a molecular weight of 620. It was identified as a derivative compound of pheophorbide structure when magne- sium comes away from a porphyrin ring. Observation of morphological changes in U937 cells follow- ing cell death induced by treated PL9443 compound revealed representative phenomena of apoptosis only in light irradiation conditions (apoptotic body, vesicle formation). Results from examining the cy- totoxicity of PL9443 substance against U937 cells showed that i nhibition rates of the cell growth were 99.9% with the concentration of 0.32 nM PL9443. Also, the caspase-3/7 activity was 99% against U937 cells with the concentration of 0.08 nM of PL9443 substance. The result of the electrophoresis was that a DNA ladder was formed by the PL9443. The PL9443 compound is a promising lead compound as a photosensitizer for photodynamic therapy of cancer.

To evaluate the effect of type of phosphate (P) material on reducing soil cadmium (Cd) extractability and radish Cd uptake in the field, two different P materials, fused superphosphate (FSP) and dipotassium phosphate (K2HPO4), were applied at rates of 0, 33.5, 100.5, and 167.5 kg P ha−1 before seeding radish (Raphanus sativa L.) in a Cd-contaminated soil. Plant-available Cd concentration in soil and total Cd uptake by radish increased with increasing FSP application but decreased with K2HPO4. The significant decrease in soil pH and negative charge of soil by FSP application led to an increase in bioavailable Cd fraction in soil and in Cd uptake by radish. However, K2HPO4 increased soil pH and negative charge of soil and decreased bioavailable Cd fractions. Soil pH and negative charge of soil played an important role in controlling Cd uptake by radish in soil to which phosphate was applied.

The objective of this study was to determine soil pH conditions that allow cadmium (Cd) to precipitate as Cd minerals in phosphate (P) amended soil. Cadmium immobilization could be attributed primarily to Cd adsorption due to increase in pH and negative charge. Soil pH might not affect Cd precipitation as Cd3(PO4)2 by direct reaction of Cd and P in the studied soil, even when soil pH increased up to 9.0. However, Cd might precipitate as CdCO3 with increasing pH up to 9.0 in P untreated soil and up to 8.0 in P treated soil depending on CO2 level.

This study was carried out to investigate changes of protease and amylase activities and nitrogen content in Chungkookjang prepared by Bacillus subtilis S8 and different soybean. Amino-type nitrogen and ammonia-type nitrogen contents increased with an increase in fermentation time and was the highest in black soybean Chungkookjang. The number of viable cells increased up to 24 h of fermentation at all temperatures tested; especially, their levels were the highest at 40. Protease activity was the highest in black soybean Chungkookjang. α-amylase activity increased significantly up to 6 h of fermentation at 30and 40and then maintained constantly. It also increased up to 30-36 h of fermentation at 45and then decreased. β-amylase activity was the highest in black soybean Chungkookjang at 35and 40and in yellow soybean Chungkookjang at 45. Production pattern of reducing sugar was similar to that of β-amylase. Amino-type nitrogen, viable cell number and reducing sugar content and β-amylase activity was the highest in Chungkookjang fermented at 40. Considering amino-type and ammonia-type nitrogen contents, Chungkookjang fermentation using yellow soybean was favorable. However, the fermentation using black soybean was favorable, considering protease and amylase activities and reducing sugar content.Key words: Bacillus subtilis, Chungkookjang, Fermentation, Soybean cultivar

Liming Effect on Cadmium Immobilization and Phytoavailability in Paddy Soil Affected by Mining Activity Chang Oh Hong, Yong Gyun Kim, Sang Mong Lee, Hyean Cheal Park, Keun Ki Kim, Hong Joo Son, Jae Hwan Cho and Pil Joo Kim (Department of Life Science and Environmental Biochemistry, Pusan National University, Miryang 627-706, South Korea, Department of Agricultural Economics, Pusan National University, Miryang 627-706, South Korea, Institute of Agriculture and Life Sciences, Gyeongsang National University, 900, Kaswa-dong, Jinju, 660-701, South Korea, Division of Applied Life Science, Graduate School (Brain Korea 21 Program), Gyeongsang National University, 900, Kaswa-dong, Jinju, 660-701, South Korea)

Adipogenesis is one of the cellular processes and a highly controlled program. Nowadays, inhibition of adipogenesis has received attention as an effective way to regulate obesity. In the current study, we investigated the inhibition effect of a chloroform extract of

The purpose of this study was to evaluate the effects of dietary Flammulina velutipes mycelium (FVM) on broiler chick performance, pathogenic bacterial (E. coli, Salmonella) counts in caecal contents and amount of NH3 in excreta. Ninety-six broiler chicks (HanHyup No. 3, Korea) were divided into four groups: 1) Control (basal diet), 2) T1 (supplemented with 1% FVM), 3) T2 (3% FVM), and T3 (5% FVM), and rose for 7 weeks. In results, there were no significant differences among treatments in weight gain, feed intake, feed efficiency and carcass yield. However, giblets were significantly increased in FVM treatments compared with control group (p

Comparison on the genomic structure and phylogenetic relationship of the Hsp88 genes from P. tenuipes Jochoen-1, P. tenuipes, C. militaris and C. pruinosa was described. The Hsp88 genes from the three entomopathogenic strains, P. tenuipes Jocheon-1(strain), P. tenuipes(original species), and C. militaris contain the identical genomic structure, namely 5 introns and 6 exons with the length of 13, 62, 32, 1,438, 306, 288 nucleotides encoding 713 amino acid residues, whereas in case of C. pruinosa, it contains 4 introns and 5 exons with the length of 13, 62, 32, 1,744, 288 nucleotides encoding 713 amino acid residues. The genomic DNA length of the Hsp88 genes from P. tenuipes Jocheon-1 and P. tenuipes are both 2,600 nucleotides long in size. The Hsp88 genes from C. militaris and C. pruinosa are 2,582, 2,576 nucleotides long in size, respectively. Hsp88 genes of the P. tenuipes Jochoen-1, P. tenuipes, C. militaris and C. pruinosa also contain the conserved ATP-binding domain. Phylogenetic analysis of the Hsp genes of the four strains tested in this study showed that the fungal Hsp88 is divided into two separate clades, ascomycetes and deutromycete. Within the ascomycetes fungal clade, the P. tenuipes Jochoen-1 and P. tenuipes formed a subgroup, on the other hand, C. militaris and C. pruinosa formed another subgroup. Pair-wise comparison of P. tenuipes Jocheon-1 Hsp88 with those of P. tenuipes, C. militaris and C. pruinosa Hsp88s revealed significant identity in deduced amino acid sequence among these strains. The P. tenuipes Jocheon-1 Hsp88 showed 99% identity with the P. tenuipes, 97% identity with the C. militaris, and 98% identity with the C. pruinosa.

In this study, we purified the extracts from the seeds and the roots of various plant species, including Q. acutissima, C. lanceolata, P. mirifica, P. bambusoides, and S. repens, and then investigated the effects of these extracts on cell growth and fat accumulation in adipocytes. We found that the extracts purified from Q. acutissima, C. lanceolata, P. mirifica, P. bambusoides, and S. repens more effectively increased the cell growth, as well as promoting the fat accumulation in adipocytes to a greater extent, than other extracts in vitro. Therefore, we made breast packs containing these effective extracts, and then investigated whether they were effective in enhancing the elasticity and volume of women’s breasts. The measurements of breast elasticity and size revealed that the breast packs efficiently increased the elasticity and size of women’s breasts. Furthermore, evaluation of the questionnaires related to usage of the breast packs indicated great satisfaction in terms of the lift, firmness, and elasticity of breasts. In conclusion, extracts purified from Q. acutissima, C. lanceolata, P. mirifica, P. bambusoides, and S. repens leading to cell growth and fat accumulation in adipocytes can effectively contribute to improving the elasticity and size of women’s breasts.

BACKGROUND: Potassium (K) is one of the macronutrients which are essential for plant growth and development. Its deficiency in paddy soils is becoming one of the limiting factors for increasing rice yield in Asia. METHODS AND RESULTS: To investigate physiological symptoms under K‐starvation (NP) compared with complete media (NPK) condition, we measured shoot/root length, weight, nutrients, and patterns of protein expression. The shoot growth was significantly reduced, but root growth was not affected by K‐starvation. However, biomasses were decreased in both shoot and root. Uptake of K was reduced up to 85%, while total concentrations of P, Ca, Mg, Na were increased in root and shoot. To better understand the starved K mechanism of rice, comparative proteome analysis for proteins isolated from rice leaves was conducted using 2‐DGE. Five spots of differentially expressed proteins were analyzed by MALDI‐TOF MS. Analysis of these K‐ starvation response proteins suggested that they were involved in metabolism and defense. CONCLUSION(s): Physiological and 2‐DGE based proteomics approach used in our study results in observation of morphology or nutrients change and identification of K‐ starvation responsive proteins in rice root. These results have important roles in maintaining nutrient homeostasis and would also be useful for further characterization of protein function in plant K nutrition.

Biochemical and enzymatic characterization for extracellular protease isolated from Cordyceps militaris cultivated on rice bran medium was investigated. C militaris produced proteolytic enzymes from 10 days after inoculation, maximum enzyme production was found at 25 days. The optimum temperature and pH of proteases production was at and pH 7.0, respectively. The protease activity was observed in the four peaks (Pro-I, Pro-II, Pro-III, and Pro-IV) separated through Sephadex G-100 column chromatography. The separated protease was optimally active at . Optimum pH of the protease was between 7 and 8. Enzyme was also stable over at . The enzyme was highly stable in a pH range of 4-9. Protease activity was found to be slightly decreased by the addition of , , , and , whereas inhibited by the addition of and Protease activity was inhibited by protease inhibitor PMSF. On the other hand, the partially purified protease was investigated on proteolytic protease activity by zymogram gel electrophoresis using three substances (casein, gelatin and fibrin). Four active bands (F-I, FII, F-III, and F-IV) of fibrin degradation were revealed on fibrin zymogram gels. Both of F-II and FIII showed caseinolytic, fibrinolytic and gelatinolytic activities in three gels. Thermostability, pH stability, and pH-thermostability of the enzyme determined the residual fibrinolytic activity also displayed on fibrin zymogram gel. The only one enzyme (F-II) displayed over a broad range of temperature at . The FII displayed fibrinolytic activity in the pH range 3-5, but was inactivated in the range of pH 6-11. The F-I and F-III showed enzyme activity in the pH range of 6-11. In the pH-thermostability, the F-II only kept fibrinolytic activity after heating at for 10, 20 and 30 min at pH 3 and pH 7, respectively. On the other hand, the F-II was retained activity until heating for 10 min under pH 11 condition. By using fibrin zymogram gel electrophoresis, extracellular fibrinolytic enzyme F-II from C. militaris showed unusual thermostable under acid and neutral conditions.

In order to study disease resistance mechanisms in rice against the rice blast fungus Magnaporthe grisea, we screened fungal elicitor-responsive genes from rice suspension-cultured cells treated with fungal elicitors employing differential hybridization (DH). By DH screening, 31 distinct rice clones were isolated and a majority of them were full-length cDNAs encoding pathogenesisrelated (PR) genes. Sixteen of the 31 genes were upregulated at 4, 8, and 12 h following fungal elicitor treatment. To elucidate the effect of signal molecules and biotic elicitors on the regulation of rice defense genes, we further characterized the transcriptional expression patterns of representative isolated PR genes; OsGlu1, OsGlu2, OsTLP, OsRLK, and OsPR-10, following treatment with fungal elicitor, phytohormones, cycloheximide, and inhibitors of protein phosphorylation. Jasmonic acid (JA) induced transcriptional expression of OsGlu1, OsTLP, and OsRLK, but not of OsGlu2 and OsPR-10 at any of the tested time points. Salicylic acid (SA) and abscisic acid weakly induced the expression of OsTLP and OsRLK. SA showed an antagonistic effect with fungal elicitor and JA. Cycloheximide suppressed all these genes upon elicitor treatment, except for OsGlu2. Staurosporine only induced the expression of OsRLK. Application of calyculin A strongly induced OsRLK expression, but suppressed the expression of OsGlu2. Our study yielded a number of PR genes that play a role in defense mechanisms against the rice blast fungus, as well as contribute towards the elucidation of crosstalk between phytohormones and other modifications during defense signaling.

The cDNA cloning and developmental profiles of the mRNA for A. pernyi arylphorin was determined. The complete A. pernyi arylphorin cDNA sequence comprised 2,234 bp (without the poly A + tail), including an open reading frame of 2,112 bp beginning with a methionine ATG at bp34. The A. pernyi arylphorin contained 704 amino acids which are highly enriched in aromatic amino acids, phenylalanine and tyrosine. The calculated molecular mass of the A. pernyi arylphorin from the ORF was 83,439 Da. The deduced amino acid sequence of A. pernyi arylphorin showed 78, 71, 62 and 64% identity with those of H. cecropia, M. sexta α subunit, M. sexta β subunit and B. mori storage protein. In Northern blot analysis, the A. pernyi arylphorin mRNA only in the fat body of the 5th instar larvae was responsible for gene expression of the protein, and the synthetic activity of the mRNA was detected strongly in the early larvae, but not in the middle or late-stage larvae. In addition, a very weak signal in mRNA activity was detected in pupal stages, but this was considered to be inactive mRNA after reviewing the results of the labeling experiment of this protein.

Cost-effective production of bacterial cellulose (BC) by Acetobacter sp. V6 was investigated in shake culture using glycerol as carbon source and its structural and physical properties were determined. In medium containing 3% (w/v) glycerol, BC production was 4.98+/-0.03g/l after 7 days. This value was 3.8-fold higher than the yield in the glucose medium. FT-IR spectra revealed that all the BC samples were highly crystalline and were cellulose type capital I, Ukrainian. The crystallinity index value of the BC produced was 9% higher in the glycerol medium than in the glucose medium. Scanning electron micrographs showed that BC from the glycerol medium was more compact than that from the glucose medium. Water-holding capacity and viscosity of BC from the glycerol medium had 61.3% and 22.4% lower values than those from the glucose medium. These results suggest that glycerol could be a potential low-cost substrate for BC production by Acetobacter sp. V6, leading to the reduction in the production cost.

We separated the bacteria showing protease activity from Reticulitermes speratus which is known as the only termite species in Korea. Then, we collected the best activated strain and studied the optimal culture condition for producing the enzyme. According to the results of observing morphological and physiological characteristics, and the type of 16S rRNA of the strain, it was identified as Serratia marcescens and named S. marcescens strain TM-3215. This strain showed the best activity under conditions of 0.8% (w/v) starch, 0.4% (w/v) peptone, 0.08% (w/v) MgSO₄?7H₂O, 30℃ and pH 8.0. After being cultivated under optimal conditions for 9 hr, the strain produced 19.8 U/ml of enzyme, an amount 1.8 times greater than the control.

The purpose of this study was to determine the effect of energy supplement on responses of plasma insulin-like growth factor (IGF)-1 and IGF binding proteins (IGFBPs) to growth hormone-releasing peptide-2 (GHRP-2) administration in normal protein-fed wethers, and to observe the effect of GHRP-2 treatment on hepatic growth hormone (GH) receptor in well-fed wethers. Plasma IGF-1 and 39-42 kDa IGFBP-3 during the HENP (CP, crude protein 0.34 and TDN, total digestible nutrients 1.83 ㎏/day DM, dry matter intake) treatment period were higher than in the LENP (CP 0.32 ㎏ and TDN 0.87 ㎏/day DM intake) period (P＜0.05). The response of GH was stimulated by GHRP-2 (12.5 ㎍/㎏ body weight/day) administration during both of the feed treatment periods (P＜0.05). The area under curve (AUC) increment and average concentration of GH (0-180 min) with GHRP-2 administration was higher during HENP treatment than LENP treatment (P＜0.01). During the HENP treatment period from day 1 to day 7 of twice daily GHRP-2 treatment, the plasma IGF-1 increment was increased on days 2, 6 and 7 of GHRP-2 administration (P＜0.05). On the basis of ligand blotting, the proportions of plasma 39-43 kDa IGFBP-3 during the HENP treatment period only showed a significant difference on days 6 and 7 with GHRP-2 administration. No significant difference in the specific binding of 125I-labeled oGH to hepatic membranes was detected between the saline and GHRP-2 treatments of the HENP-fed wethers. These results suggest that the nutritional balance between energy and protein may affect the endogenous GH / IGF-1 axis as well as plasma IGFBP-3 levels.

The occurences of proteins relating to Antheraea pernyi arylphorin in haemolymph, fat body, integument, midgut and silkgland of the wild silkmoths, Antheraea yamamai, Antheraea pernyi, Samia cynthia pryeri and Actias gnoma in the 5th larval instar were investigated by immunoblot analysis using mouse polyclonal antibody against A. pernyi arylphorin as probe. In A. yamamai, A. pernyi, S. cynthia pryeri and A. gnoma, the major immunoreactive antigenic proteins with a molecular weight of 80 KDa against the antisera of the A. pernyi arylphorin were clearly observed in the haemolymph, but in the integument, fat body, midgut and silkgland of the corresponding wild silkmoths the presence of the immunoreactive proteins were very variable. These results suggest that the A. pernyi arylphorin has almost same immunological identity with those of the wild silkmoths, A. yamamai, S. cynthia pryeri and A. gnoma though the distribution of the corresponding antigenic arylphorins is different according to the tissues of the wild silkmoths.

A mineral phosphate solubilizing bacterium, Burkholderia cepacia DA23 has been isolated from cultivated soils. Phosphate-solubilizing activities of the strain against three types of insoluble phosphate were quantitatively determined. When 3% of glucose concentration was used for carbon source, the strain had a marked mineral phosphate-solubilizing activity. Mineral phosphate solubilization was directly related to the pH drop by the strain. Analysis of the culture medium by high pressure liquid chromatography identified gluconic acid as the main organic acid released by Burkholderia cepacia DA23. Gluconic acid production was apparently the result of the glucose dehydrogenase activity and glucose dehydrogenase was affected by phosphate regulation.Uma bactéria capaz de solubilizar fosfato mineral, Burkholderia cepacea DA23, foi isolada de solo cultivado. A capacidade dessa bactéria solubilizar o fosfato de três tipos de fosfato insolúvel foi quantificada. Quando foi utilizada glicose a 3% como fonte de carbono, a bactéria apresentou uma intensa atividade solubilizante de fosfato, sendo a solubilização diretamente relacionada com a queda de pH causada pela bactéria. A análise do meio de cultura por cromatografia líquida de alta pressão indicou o ácido glicônico como principal ácido produzido por Burkholderia cepacea DA23. Aparentemente, a produção de ácido glicônico foi causada pela atividade da glicose desidrogenase. A enzima foi afetada pela regulação do fosfato.

A mineral phosphate solubilizing bacterium, Burkholderia cepacia DA23 has been isolated from cultivated soils. Phosphate-solubilizing activities of the strain against three types of insoluble phosphate were quantitatively determined. When 3% of glucose concentration was used for carbon source, the strain had a marked mineral phosphate-solubilizing activity. Mineral phosphate solubilization was directly related to the pH drop by the strain. Analysis of the culture medium by high pressure liquid chromatography identified gluconic acid as the main organic acid released by Burkholderia cepacia DA23. Gluconic acid production was apparently the result of the glucose dehydrogenase activity and glucose dehydrogenase was affected by phosphate regulation.

This study estimated the mound litter biomass and the nutrient content of mound litter in the two study plots in Durokoppin Nature Reserve, Western Australia between 2004 and 2005. There were no significant differences in biomass of litter in individual mounds between the two study plots. Seven components of litter were found in the mounds. The nutrient concentrations were higher in the wood-land than in the shrubland plot, although the differences were not statistically tested, and the total amount of each nutrient measured was generally greater in the woodland than in the shrubland plot. The aforementioned results show that D. tamminensis harvests various plant material according to bio-mass availability. The role of this termite takes on particular importance in view of the fact that Western Australian soils are notoriously impoverished in nutrients.

To develop multifunctional microbial inoculant, microorganisms with antagonistic activity and biofertilizing activity were screened. Pantoea agglomerans and Bacillus megaterium from our laboratory culture collection, and strain MF12 from soil near poultry farm in Miryang were selected. On the basis of morphological, physiological studies and 16S rDNA sequence analysis, isolate MF12 was identified as the Bacillus pumilis. Three strains were studied for insoluble phosphate solubilization, indole-3-acetic acid (IAA) and siderophore production, ammonification ability, hydrolytic enzyme production and antifungal activity against phytopathogenic fungi. P. agglomerans did not produce any visible clear zone on agar plate containing 0.5% as a sole phosphorus source. However, this strain could solubilize insoluble phosphate in liquid medium. All strains produced IAA ranged from depending on culture time and had ammonification ability. Among three strains, only P. agglomerans produced siderophore. P. agglomerans produced pectinase and lipase, B. megaterium produced amylase, protease and lipase while B. pumilis produced protease and lipase. P. agglomerans showed antifungal activities against phytopathogenic fungi, Fusarium oxysporum and Colletotrichum gloeosporioides. B. pumilis showed antifungal activities against Botrytis cinerea, Sclerotinia sclerotiorum and Phythium ultimum.

The created microbial pesticide was used to examine its effects using the inside-pot test method. The selected microbial pesticide KBC3017 particle pesticide manufactured by using Monacrosporium thaumasium was used in the farm-house outdoor test to find the optimum consistency and its effects. The more amount used, the better effect it showed. However, the optimum consistency was 2% and the KBC3017 particle pesticide for which the diatomite and raw jade powder were used as an increaser, when used 2% level of the total amount of soil, showed 71% effect on nematode prevention. The root and the stem of crops were better compared to those without any pesticide used.

The fruits of Zanthoxylum piperitum are known as having various physiology vitality, and the abstraction ingredient of the pericarp is also known as having strong antibiotic activities against various bacteria. Therefore, this study was carried out to estimate the effect of physiology vitality when the abstraction ingredient of Z. piperitum was added in soy sauce(Chopi-kanjang) and soybean paste(Chopi-doenjang). For the antibiotic activity against the pathogens of sitotoxism such as Staphylococcus aureus, Salmonella typhimurium, Vibrio parahemolyticus, Escherichia coli 0157:H7, the extracts of the Chopi-kanjang was added 1%, 2%, 4% pericarp of Z. piperitum in the manufacturing process of soy source. According to the results, the growth of E. coli 0157:H7 and V. parahemolyticus were respectively inhibited as 70% and 50% by the Chopi-kanjang added 2% of the ingredient. For the antibiotic effects of the aforementioned Chopi-kanjang against Sal. typhimurium and Sta. aureus, the growth of those pathogens was also inhibited between 40% and 60% according to the manufacturing period of Chopi-kanjang. It was confirmed that the antibiotic activity using the mixture of the abstraction ingredient and Chopi-doenjang was lower than those of Chopi-kanjang. In order to estimate the anticancer activity using by caspase-3 activity, the mixture of the abstraction ingredient of the pericarp of Z. piperitum and Chopi-kanjang was treated to leukemia cells. According to the results, the activities of caspase-3 using the mixture added 1%, 2% and 4% of the abstraction ingredient were respectively increased as much as 4, 12, 15 times comparing with the control which was treated with the soy source only. It could be that the mixture of the abstraction ingredient of the pericarp of Z. piperitum and soy source induced apoptosis, and the mixture of the abstraction ingredient and soybean paste had no effect on the activity of caspase-3. In order to find out the death of the aforementioned cells caused by necrosis or apoptosis, DNA fragmentation in the cell was examined. U-937 cells showed apoptotic DNA fragmentation in the incubation with Chopi-kanjang extract.

A total of 5 female elk deer() were included in a study on the changes in physico-chemical properties of deer meat during storage at . The deers were exposed to normal pre-slaughter handling and put under anesthesia before slaughtered. The loin and leg cuts were deboned from the carcass after 24hrs slaughter. The samples weighing approximately 300g were packaged using wrap packaging and stored for 1, 2, 3, 4, 5 and 6 months at . During the freezing period, the changes of moisture maintenance was slower for the meats kept at than that at . The meat softness represented higher value after freezing, and it tended to be decreased with the passage of storage time. Comparing the values between freezing temperature, the value at was higher than at . All meats frozen at were edible after storage for 6 months. Loin was inedible, and leg was edible after freezing storage for 6 months. The pH values of the meats were increased during freezing period, even though pH value of the meats stored for 6 months was less than 5.70. Luminosity of loin was significantly increased with the passage of storage time, although loin stored at was darker, and leg stored at was darker. The index of red color was rapidly decreased for loin stored at , and for leg stored at , respectively. There were no changes in the index of yellow color for loin stored at , and the index for leg was increased with regardless of the freezing storage temperature after 6 months.

Lignin degrading bacterium Pseudomonas sp. LG2 was able to degrade lignin substrate to a lot of APPL compound. APPL compound was detected in culture supernatants from Pseudomonas sp. LG2 grown with BSC(brewer`s spent grain). FAE(ferulic acid esterase) and xylanase are induced from Pseudomonas sp. LG2 in the presence of carbon sources such as oat spelt xylan, HBSG I, II(hydrolyzed brewer`s spent grain I, II) and AFBSG(autoclaved fraction from brewer`s spent grain). However, xylanase and FAE are not induced by growth of Pseudomonas sp. LG2 on xylose and arabinose. Pseudomonas sp. LG2 is grown on medium containing oat spelt xylan, HBSG I, II and AFBSG and the induction of FAE and xylanase activities of extracellular proteins determined during 14 days. Maximum level of xylanase activity(5.3 U/mg) found at 6 days in culture contained oat spelt xylan as carbon source, whereas maximum level of FAE activity(15.4 mU/mg) was found at 8 days in culture contained AFBSG as carbon source. Most ferulic acid was released in culture supernatants when Pseudomonas sp. LG2 grown on oat spelt xylan, HBSG I, II and AFBSG. FAE of extracellular enzymes was also specific activity on methyl ferulic acid, methyl caffeic acid and methyl p-coumaric acid respectively, but not methyl sinapinic acid, methyl vanillic acid and methyl gallic acid.

A total of 5 female elk deer (220 kg) were included in a study on the changes in physico-chemical properties of deer meat during storage at . The deers were exposed to normal pre-slaughter handling and put under anesthesia before slaughtered. The loin and leg cuts were deboned from the carcass after 24hrs slaughter. The samples weighing approximately 300 g were packaged using three packaging methods : Atmospheric packaging(A), Vacuum packaging (V) and Modified atmospheric at packaging (M: 80% +20% )and stored for 3, 7, 11, 15, 20 and 25days at . The ability of moisture maintenance tended to be decreased with the passage of storage time, and that of the meats treated with the vacuum wrapping was lower than that with other wrapping methods. TBARS values of the meats with the regardless of treatments was increased with the passage of storage time, although the values with vacuum wrapping was lowest. VBN values of the meats with the regardless of treatments was also increased with the passage of storage time, although the values with vacuum wrapping was also lowest. The pH values of the meats with the regardless of treatments was also increased with the passage of storage time, although the values with vacuum wrapping was also lowest. For the meat softness, the value of the meats treated with atmospheric packaging was increased with the passage of storage time, but the values with vacuum wrapping and modified atmospheric packaging were respectively decreased. There were no significant differences in luminosity of leg, although loin treated with vacuum wrapping was significantly brighter than that with atmospheric packaging and modified atmospheric packaging. In general, the index of red color of the meats treated with vacuum wrapping was higher than that with atmospheric packaging and modified atmospheric packaging. The index of yellow color tended to be decreased after increment for a time with the passage of storage time, although the value with vacuum wrapping was generally higher than that with other wrapping methods.