• The Korean Journal of Pesticide Science
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• v.17 no.4
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• pp.379-387
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• 2013

In order to improve practical utility of agro-microorganisms (AMs) which had been cultured and disseminated to promote plant growth and to control crop diseases, 51 isolates of AMs were collected from 18 agricultural extension centers in local government and screened for multi-functions such as antifungal activity, activities of phosphorus solubilization, IAA and siderophore production, nitrogen fixation, and hydrolytic enzyme activity. Finally we selected one isolate showing good antifungal activity and multi-functions related to plant growth and disease control. The selected isolate, Paenibacillus polymyxa CW, showed good inhibitory effect against plant pathogens, Pyricularia gresea, Colletotrichum acutatum, Fusarium oxysporum, Phomopsis sp., Aspergillus niger, Rhizoctonia solani and Phytophthora capsici. Suppressive effect of P. polymyxa CW against the used plant pathogens except for R. solani was much higher than that of P. polymyxa AC-1 storing in National Academy of Agricultural Science. We found P. polymyxa CW isolate showed good activity in siderophore and IAA formation, and nitrogen fixation. With P. polymyxa CW isolate, siderophore formation activity was similar to that of P. polymyxa AC-1, but IAA formation and nitrogen fixation activity was much higher than that of P. polymyxa AC-1. However neither P. polymyxa CW nor P. polymyxa AC-1 showed hydrolytic enzyme (chitinase, pectinase and cellulase) activity. The treatment of P. polymyxa CW with culture suspension of different cell density ($10^8$, $10^7$. $10^6$ cfu/ml) showed that the highest density reduced incidence of red pepper powdery mildew by 68.3% after 10 days of application. As application density of P. polymyxa CW was decreased, its control efficacy was proportionally decreased. In addition, when P. polymyxa CW was treated to control tomato powdery mildew at the same concentrations and their control effects were investigated after 7 days of inoculation, disease incidence was 0.03, 19.5, 45.7%, respectively, compared to 56.3% that of untreated check. Like red pepper powdery mildew, increase of application density of P. polymyxa CW resulted in increase of its control efficacy proportionally. P. polymyxa CW showed a density-dependent control efficacy against red pepper and tomato powdery mildews. Therefore we think that mode of action of the antagonist for suppressing two powdery mildew diseases might be antibiosis and density of more than $10^8cfu/ml$ was needed to control effectively the two diseases. On this basis, we think that P. polymyxa CW can be a promising control agent for suppressing powdery mildews of red pepper and tomato.

• Journal of Life Science
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• v.12 no.6
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• pp.700-707
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• 2002

To study the characteristics of organic and nutrient removal by Bacillus species at high COD concentration of influent, three lab-scale batch reactors(R1, R2, R3), each of which has different substrate composition, were operated. More than 95% of $NH_4^+$-N and $COD_{cr}$, concentrations were removed under an aerobic condition, and their removal efficiencies were found to be 22.6 and 90.5%(R1), 23.9 and 65.8%(R2), 30.2 and 86.4%(R3), respectively. The removal efficiency of $NH_4^+$-N was high when an enough amount of $NO_3^{-}$-N was supplied, and that of $COD_{cr}$. was low when a high concentration of initial $NO_2^{-}$-N was added. The amount of carbon utilized in denitrification was a little. In all reactors,$NO_3^{-}$-N was removed under an anoxic condition, but in the R3 reactor, 10% of $NO_3^{-}$-N could be removed even undo, an aerobic condition. The removal efficiencies of TN and TP were 41.8 and 49.5%(R1), 40.1 and 35.8%(R2), 47.0 and 57.6%(R3), respectively. Alkalinities destructed under an aerobic condition for each reactor were 4.96, 5.41 and 3.93 mg/L (as $CaCO_3$) per each gram of $NH_4^+$-N oxidized, respectively, while 3.06, 3.17 and 2.60 mg/L (as $CaCO_3$) of alkalinities were produced for each gram of ,$NO_3^{-}$-N reduced to $N_2$. The SOUR were found to be 38.5, 52.7 and 42.0 mg $O_2$/g MLSS/hr, which indicated that Bacillus sp. had a higher cell activity than activated sludge. The OLR and sludge production were estimated to be 0.69 and 0.28(Rl), 0.77 and 0.20(R2), 0.61 kg COD/$m^3$/day and 0.25 kg MLSS/kg COD(R3), respectively. From the N-balance, the highest percentage(40.9%) of nitrogen lost to $N_2$ was obtained in the R3 reactor. From all the results, the possibility of aerobic denitrification Bacillus sp. has been shown and the B3 process seemed to have two advantages: a little amount of carbon was required in denitrification and not much amount of alkalinity was destructed under an aerobic condition.

• Korean Journal of Animal Reproduction
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• v.22 no.4
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• pp.425-435
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• 1998

To improve the efficiency of nuclear transplantation in bovine, in this study the development in vitro of nuclear transferred (NT) embryos was compared by different activation regimens of the enucleated oocytes. The effect of developmental stage and culture system of donor nuclei on fusion and development in vitro of NT embryos were also evaluated. Oocytes were collected from Hanwoo ovaries obtained from slaughterhouse and matured in Ham's F-10 supplemented with hormones. After 20~22 h maturation, the oocytes were vortexed to be free from cumulus cells and subsequently their nucleus and the first polar body were removed. Enucleated oocytes were divided into 3 groups for activation; the oocytes of group I were activated with ionomycin for 5 min and subsequently incubated in 6-dimetylarninopurine (DMAP) for 4 h, Those of group II were treated with DMAP for 4 h at 39 h after onset of in vitro maturation (IVM) and those of group III were kept in room temperature ($25^{\circ}C$) for 3 h at 39 h after onset of IVM. After in vitro fertilization (IVF) the embryos for muclear donor were cultured either by group culture (20 embryos /50 ${mu}ell$ drop) or individually (1 embryo /50 ${mu}ell$ drop) for 4 day and 5 day. At day 4 and 5 after IVF, blastomeres were separated in calcium-magnesium free medium, and then classified into small (day 5: $\leq$ 38 ${\mu}{\textrm}{m}$, day 4: $\leq$ 46 ${\mu}{\textrm}{m}$) and large (day 5 : $\geq$ 38 ${\mu}{\textrm}{m}$, day 4 ; $\geq$ 46 ${\mu}{\textrm}{m}$). The separated blastomeres were replaced into enucleated and activated recipient cytoplasm. The blastomere-oocyte complexes were fused by electrically. The NT embryos were cultured in TCM-199 containing 10% FCS in 39$^{\circ}C$, 5% $CO_2$ incubator for 7 day. The results obtained were summarized as follows; There were no differences in fusion and development to blastocyst between groups as group I (68%, 10%), group II (75%, 14%) and group III (73%, 9%), respectively. However, the cell number in blastocyst of NT embryos in group III were significantly fewer than in the other groups (P＜0.05). No differences in fusion and development to blastocyst were found between individual or group cultured and between small or large blastomeres of day 4 and day 5 donor embryos. From these results, it was concluded that the combination of ionomycin and DMAP, or treatment of DMAP at 39 h after onset of IVM were useful for the efficient of production of NT bovine embryos, and the individual cultured embryos could be simply used as donor nuclei for NT bovine embryo.

• Food Science of Animal Resources
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• v.30 no.6
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• pp.975-988
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• 2010

Two experiments were conducted to determine whether water extracts from Artemisia capillaries (A. capillaries) and Camellia sinensis (C. sinensis) could be used as alternatives to antibiotic growth promoters in broiler feed. The experiment 1 was verified their chemical composition, extracts yields, total phenolic compounds concentration, antioxidant activity, antimicrobial activity, and chicken splenocytes proliferation through in vitro test. The extract yields of A. capillaries and C. sinensis were 26.5 and 16.8%, respectively. Total phenolic compounds concentrations of them expressed as gallic acid equivalent were 15.28 and 26.74 mg/mL, respectively. Electron donating abilities of them expressed as $SC_{50}$ showing 50% DPPH radical scavenging were 0.30 and 0.06 mg, respectively. Bacterial inhibitory rates of them against Escherichia coli, Staphylococcus aureus, and Salmonella Typhimurium were ranged from 42.1 to 52.3% and from 21.6 to 33.7%, respectively. And, these extracts increased proliferation of chicken splenocytes. Especially, A. capillaris was more excellent than Echinacea and Concanavalin A known as T-cell stimulator. The experiment 2 was investigated their effects on growth performance, relative organ weight, cecal microflora, blood biochemical parameters, and splenic cytokines mRNA expression in broiler chicks. Four hundred eighty 1-day-old male broiler chicks (Ross 308) were divided in to 4 treatment groups with 4 replicates of 30 birds in each group: NC (control, no antibiotics), PC (avilamycin, 10 ppm; salinomycin, 60 ppm), AC (A. capillaries, 100 ppm), and CS (C. sinensis, 100 ppm); treatments were administered through water supplementation. Final body weight was significantly higher in all treated groups than in NC (p<0.05). Cecal Salmonella numbers were significantly or somewhat decreased in all treated groups than in NC (p<0.05). The relative weights and lengths of the small intestine were more significantly decreased in the PC and AC groups than in the other groups. Cecal Salmonella numbers were significantly or somewhat decreased in all treated groups than in the NC group (p<0.05). The contents of total cholesterol, aspatate aminotransferase, and alanine aminotransferase in blood serum were more significantly decreased in all treated groups than in NC (p<0.05). In conclusion, these results suggested the possibility that these extracts could serve as alternatives for antibiotic growth promoters.

• The Journal of Natural Sciences
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• v.4
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• pp.103-142
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• 1991

These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

• Korean Journal of Agricultural and Forest Meteorology
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• v.8 no.1
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• pp.1-9
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• 2006

Regardless of the recent observed warmer winters in Korea, more freeze injuries and associated economic losses are reported in fruit industry than ever before. Existing freeze-frost forecasting systems employ only daily minimum temperature for judging the potential damage on dormant flowering buds but cannot accommodate potential biological responses such as short-term acclimation of plants to severe weather episodes as well as annual variation in climate. We introduce 'dormancy depth', in addition to daily minimum temperature, as a complementary criterion for judging the potential damage of freezing temperatures on dormant flowering buds of grape vines. Dormancy depth can be estimated by a phonology model driven by daily maximum and minimum temperature and is expected to make a reasonable proxy for physiological tolerance of buds to low temperature. Dormancy depth at a selected site was estimated for a climatological normal year by this model, and we found a close similarity in time course change pattern between the estimated dormancy depth and the known cold tolerance of fruit trees. Inter-annual and spatial variation in dormancy depth were identified by this method, showing the feasibility of using dormancy depth as a proxy indicator for tolerance to low temperature during the winter season. The model was applied to 10 vineyards which were recently damaged by a cold spell, and a temperature-dormancy depth-freeze injury relationship was formulated into an exponential-saturation model which can be used for judging freeze risk under a given set of temperature and dormancy depth. Based on this model and the expected lowest temperature with a 10-year recurrence interval, a freeze risk probability map was produced for Hwaseong County, Korea. The results seemed to explain why the vineyards in the warmer part of Hwaseong County have been hit by more freeBe damage than those in the cooler part of the county. A dormancy depth-minimum temperature dual engine freeze warning system was designed for vineyards in major production counties in Korea by combining the site-specific dormancy depth and minimum temperature forecasts with the freeze risk model. In this system, daily accumulation of thermal time since last fall leads to the dormancy state (depth) for today. The regional minimum temperature forecast for tomorrow by the Korea Meteorological Administration is converted to the site specific forecast at a 30m resolution. These data are input to the freeze risk model and the percent damage probability is calculated for each grid cell and mapped for the entire county. Similar approaches may be used to develop freeze warning systems for other deciduous fruit trees.

• Journal of Embryo Transfer
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• v.29 no.1
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• pp.73-81
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• 2014

The in vitro production of porcine embryos was essential to increase of blastocyst development rate and select of high quality blastocyst in early stage. There were a lot of reports about in vitro porcine embryo development, but there was no report about the selection of high quality embryos. Therefore, in this study, we investigated the effect of vitamin $K_1$ (vit $K_1$) on the development and survival rate of porcine in vitro fertilized embryos. When vit $K_1$ was treated for 24 hr at day 1 in vitro culture, blastocyst development rate in the control group ($35.5{\pm}3.2%$) was significantly lower compared to $1.0{\mu}M$, $3.0{\mu}M$, or $6.0{\mu}M$ groups ($14.5{\pm}4.3$, 0.0, or 0.0%; p<0.05). The survival rates of blastocysts at day 8 in $1.0{\mu}M$, $3.0{\mu}M$ or $6.0{\mu}M$ of vit $K_1$ treated groups ($22.2{\pm}2.9$, 0.0 or 0.0%) were significantly lower than that of the control group ($31.8{\pm}2.6%$; p<0.05). We were added at $1.0{\mu}M$, $3.0{\mu}M$ or $6.0{\mu}M$ vit $K_1$ for different durations of time at day 1 in vitro culture. The development rate and survival rate in the group of $1.0{\mu}M$ vit $K_1$ for 6 hr was $26.5{\pm}2.9%$ and $47.2{\pm}2.8%$, respectively, which were differed significantly in the group of 12 hr (p<0.05). In the group of $3.0{\mu}M$ vit $K_1$, the blastocyst development in control group was $36.4{\pm}3.1%$ but, the survival rate $41.7{\pm}3.2%$ in the group of 3.0 hr was significantly higher than that of the control group (p<0.05). In the group of $6.0{\mu}M$ vit $K_1$, the control group's the blastocyst development was $32.0{\pm}2.8%$ and the 0.5 hr supplement group's survival rates was $42.9{\pm}1.8%$ higher than other groups. We added vit $K_1$ at day 1, day 2, day 4 and day 6 of in vitro culture, on the based the results of supplemented concentration and duration. In the group of $1.0{\mu}M$ 6.0 hr addition, the blastocyst development rate of day 4 and the survival rate of day 2 were the highest in each group. In the groups of $3.0{\mu}M$ 3.0 hr addition or $6.0{\mu}M$ 0.5 hr addition, the blastocyst development ($59.5{\pm}4.1%$ and $50.0{\pm}3.6%$) and survival rates ($72.7{\pm}5.4%$ and $79.2{\pm}4.0%$) on day 4 were significantly higher than that of control and other experiment groups (p<0.05). Meanwhile, the number of cells in blastocysts that produced by vit $K_1$ supplementation was $53.4{\pm}5.8$, $49.4{\pm}3.8$ and $51.5{\pm}4.5$ respectively, which were significantly higher than that of $40.2{\pm}2.3$ in the control group (p<0.05). There was no difference of the number of apoptotic cells between control and experiment groups. In addition, gene expression of survival blastocyst, the Bax mRNA expression was similar between the control and the experiment groups. However, Bcl-xL mRNA expression's in the group of $6.0{\mu}M$ 0.5 hr on day 4 was highest among control and experiment groups (p<0.05). In this study suggested that the control of concentration, duration and time was effective on the survival and cell number of porcine blastocyst derived from in vitro. We are not know what the exact reasons of the effect of vit $K_1$ on embryo development and need to fur ther study. However, vit $K_1$ might be using the selection of high quality porcine blastocyst.

• Tuberculosis and Respiratory Diseases
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• v.45 no.4
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• pp.861-869
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• 1998

Backgrounds: The injury of a tissue results in the infalmmation, and the imflammed tissue is replaced by the normal parenchymal cells during the process of repair. But, constitutional or repetitive damage of a tissue causes the deposition of collagen resulting in the loss of its function. These lesions are found in the lung of patients with idiopathic pulmonary fibrosis, complicated fibrosis after diffuse alveolar damage (DAD) and inorganic dust-induced lung fibrosis. The tissue from lungs of patients undergoing episodes of active and/or end-stage pulmonary fibrosis shows the accumulation of inflammatory cells, such as mononuclear cells, neutrophils, mast cells and eosinophils, and fibroblast hyperplasia. In this regard, it appears that the inflammation triggers fibroblast activation and proliferation with enhanced matrix synthesis, stimulated by inflammatory mediators such as interleukin-1 (IL-1) and/or tumor necrosis factor (TNF). It has been well known that TGF-$\beta$ enhance the proliferation of fibroblasts and the production of collagen and fibronectin, and inhibit the degradation of collagen. In this regard, It is likely that TGF-$\beta$ undergoes important roles in the pathogenesis of pulmonary fibrosis. Nevertheless, this single cytokine is not the sole regulator of the pulmonary fibrotic response. It is likely that the balance of many cytokines including TGF-$\beta$, IL-1, IL-6 and TNF-$\alpha$ regulates the pathogenesis of pulmonary fibrosis. In this study, we investigate the interaction of TGF-$\beta$, IL-1$\beta$, IL-6 and TNF-$\alpha$ and their effect on the proliferation of fibroblasts. Methods: We used a human fibroblast cell line, MRC-5 (ATCC). The culture of MRC-5 was confirmed by immunofluorecent staining. First, we determined the concentration of serum in cuture medium, in which the proliferation of MRC-5 is supressed but the survival of MRC-5 is retained. Second, we measured optical density after staining the cytokine-stimulated cells with 0.5% naphthol blue black in order to detect the effect of cytokines on the proliferation of MRC-5. Result: In the medium containing 0.5% fetal calf serum, the proliferation of MRC-5 increased by 50%, and it was maintained for 6 days. IL-1$\beta$, TNF-$\alpha$ and IL-6 induced the proliferation of MRC-5 by 45%, 160% and 120%, respectively. IL-1$\beta$ and TNF-$\alpha$ enhanced TGF-$\beta$-induced proliferation of MRC-5 by 64% and 159%, but IL-6 did not affect the TGF-$\beta$-induced proliferation. And lNF-$\alpha$-induced proliferation of MRC-5 was reduced by IL-1$\beta$ in 50%. TGF-$\beta$, TNF-$\alpha$ and both induced the proliferation of MRC-5 to 89%, 135% and 222%, respectively. Conclusions: TNF-$\alpha$, TGF-$\beta$ and IL-1$\beta$, in the order of the effectiveness, showed the induction of MRC-5 proliferation of MRC-5. TNF-$\alpha$ and IL-1$\beta$ enhance the TGF-$\beta$-induced proliferation of MRC-5, but IL-6 did not have any effect TNF-$\alpha$-induced proliferation of MRC-5 is diminished by IL-1, and TNF-$\alpha$ and TGF-$\beta$ showed a additive effect.