• Korean Journal of Environmental Agriculture
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• v.24 no.3
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• pp.245-252
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• 2005

The objective of the present work was to determine the corresponding uptake and assimilation of ${NO_3}^-$ in roots and shoots of leafy radish by applying of mixed amino acids (MAA). The amino acids used in this experiment were alanine (Ala), ${\beta}-alanine\;({\beta}-Ala)$, aspartic acid (Asp), asparagines (Asn), glutamic acid (Glu), glutamine (Gln), and glycine (Gly). Leafy radish was grown by conventional fertilization with macro- and micronutrients under controlled conditions. The 15-day-old seedlings were treated 0, 0.3 and 3.0 mM of MAA containing 5 mM ${NO_3}^-$ in growth medium. Nitrate uptake was determined by following ${NO_3}^-$ depletion from the uptake solution. The activity of the enzymes related to the process of ${NO_3}^-$ reduction (NR: nitrate reductase; NiR: nitrite reductase; GS: glutamine synthetase) and the content of ${NO_2}^-\;and\;{ND_3}^-$ were analyzed in shoots and roots. The results of this study showed that ${NO_3}^-$ uptake was inhibited 38% with treatment of 0.3 mM of MAA. However, there was more than three times increase of N03- uptake in 3.0 mM MAA. In addition, the enzymatic activities were positively affected by the high MAA rate. Finally, the ${NO_3}^-$ content was increased slightly both in shoots and roots of leafy radish by MAA treatments.

• Horticultural Science & Technology
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• v.30 no.4
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• pp.357-362
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• 2012

Proper light intensity, shading tolerance, potting media, and fertilization level were investigated to develop Orostachys iwarenge for. magnus (Korean name, Ullungyeonwhabawisol) as a potted ornamental plant. The plants were grown under different light intensity (52, 82, 90, and 97% shading). The best growth was shown at 52% shading, which indicated that the proper light intensity for O. iwarenge for. magnus is less than 52% of shading. Plant growth decreased severely at 82% shading and leaf color became lighter as the shading rate increased, which indicated that O. iwarenge for. magnus has no tolerance against low light intensity. To select a proper potting media, decomposed granite (DG), fertilizer-amended media (FAM), river sand (RS) were used as potting medium with different ratio of 60:20:20 (DG:FAM:RS, v/v/v), 80:20 (DG:FAM, v/v), 60:40 (DG:FAM, v/v), and 20:80 (FAM:RS, v/v). DG:FAM:RS (60:20:20) showed the highest values in shoot fresh weight, plant width, and number of runner in potted O. iwarenge for. magnus. Fresh weight of shoot part was 16 g in DG:FAM:RS (60:20:20), which was about 2 folds of those at the other medium. At the experiment for selection of proper fertilization level, plants showed a better growth as the concentration of hyponex solution and application frequency increased. Once drenching 1 week interval of hyponex solution diluted by 1,000 folds brought the highest results in fresh weight, plant width, and runner number. Particularly, fresh weight of shoot part was 35 g at once drenching per week of 1,000 folds solution, indicating 84% improvement comparing with non treatment (19 g).

• Journal of Korean Society of Forest Science
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• v.35 no.1
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• pp.1-8
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• 1977

Forest soil fertility was evaluated through the approach of numerical method. In this study, the soil chemical properties analyzed for 35 different soil series as table 2 were cited in numerical analysis. Minimum contents of essential nutrients in the surface soil for a satisfactory growth of tree in the plantation were evaluated by comparing with Wild's standard as table 1. Demanding level of fertilization were evaluated by using the formula 1 as table 5. Similar relation of soil chemical properties between soil series were calculated through formula 2, and then classified into 5 groups in soil chemical properties. 1. General chemical properties of surface soil in case of 35 soil series. About 40 percent of 35 different soil series are less than 2 percent in organic matter, 10 ppm in available phosphorus, 1.25m.e/l00g in exchangeable calcium and 0.5m.e/l00g in exchangeable magnesium. Generally, shortage of exchangeable potash are not found. CEC less than 10m.e/l00g are in two thirds and strong acid soil less than PH 5.5 are in about four fifths. 2. Soil series requested or not the fertilization are indirectly evaluated from the formula 1 using the relative figure of chemical components of CEC, OM and MgO. Through this analysis, 8 different soil series have very poor quality in soil chemical capacity so that demands highly the fertilization. On the other hand, other 13 different soil series group have not been thought to need the fertilization according to chemical guality. 3. By the results comparing the similarity of chemical properties of forest soil, it is thought to be suitable that the forest soil fertility are divided into 5 groups as follows: 1. Low CEC soil 1-1 Low organic matter soil less than 2 percent 1-2 Medium organic matter soil less than 4 percent 2. High CEC and organic matter soil 2-1 Low magnesium soil 2-2 High magnesium soil 3. High magnesium and calcium soil as lime stone.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.223-228
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• 2002

This study was carried out to examine the effects of antioxidants and buthionine sulfoximide(BSO) on the development of porcine in vitro maturation(IVM) and in vitro fertilization(IVF) oocytes. Cumulus cell free embryos derived from porcine IVM/IVF oocytes were cultured NCSU23 medium with antioxidants or BSO under an atmosphere of 5% $CO_2$ and 5% $O_2$at 38.5$^{\circ}C$ for 5~6 days. The embryos cultured in medium with BSO showed a significanthy(P<0.05) lower rates(8.4~15.7%) of the development to the morulae and blastocyst stage than control group(35.9%). When the embryos were cultured with NAC, ebselen, glutathione and BSO, the proportions of embryos beyond morulae and blastocysts were significantly(P<0.05) higher in medium with NAC(40.5%), ebselen(44.2%) and glutathione(36.0%) than BSO(10.9%). In conclusions, these results indicate that NAC, ebselen and glutathione as a antioxidants can increase the proportion of embryos that develop beyond morulae stage. BSO, intracellular glutathione inhibitors, is suppressed the development of porcine embryos derived from IVM/IVF.

• Journal of Embryo Transfer
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• v.17 no.3
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• pp.211-218
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• 2002

These studies were carried out to investigate the effects of the addition of amino acids and FBS as source of exogenous nitrogen fixation added to medium on in vitro production of blastocyst derived from bovine follicular oocytes. The base medium was TCM-199 solution for in vitro maturation(IVM) of bovine follicular oocytes and Fer-TALP solution for in vitro fertilization(IVF) and YS solution for in vitro culture(IVC). IVC used the fertilized oocytes of 24-hr culture (day 1) after IVF. Rmbryos were cultured in drop-culture that contained 25 embryos per 10 ${mu}ell$. The results obtained are as follows: 1 The developmental rates of fertilized oocytes to blastocyst that developed from YS solution with NEAA derived from MEM alone were higher than those of YS solution without NEAA. 2. The developmental rates of fertilized oocytes to blastocyst that developed from YS solution with EAA derived RPMI 1640 alone were significantly higher than those of YS solution without EAA (p<0.05). 3. When added to EAA on day 5 after NEAA supplementation on day 1, the developmental rates of hatched blastocyst and blastocyst to hatched blastocyst were improved. 4. When removed to EAA on day 3, day 4 and day 5 from medium after NEAA and EAA supplementation on day 1. the developmental rates of blastocyst to hatched blastocyst were reduced. 5. When added to FBS as source of exogenous nitrogen fixation, the developmental rates of blastocyst and hatched blastocyst that developed from the later culture higher(day 5) than those of the early culture.

• Journal of Embryo Transfer
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• v.23 no.4
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• pp.269-274
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• 2008

The purpose of this study was to determine the effects of Taxol pre-treatment to in vitro matured bovine oocytes, and sucrose and trehalose added to vitrification solution on spindle morphology and embryonic development following cryopreservation. Bovine oocytes were collected from ovaries and matured in tissue culture medium 199 (TCM 199) supplemented with 10% Fetal Bovine Serum (FBS), 0.05ng/ml epidermal growth factor, 0.01 IU/ml luteinizing hormone and $1{\mu}g/ml$ estradiol for 22h in $39^{\circ}C$, 5% $CO_2$, TCM 199-HEPES containing 20% FBS was used as basic medium (BM) to prepare vitrification solution. Oocytes were pre-treated with $1\;{\mu}M$ Taxol in maturation medium for 15 min prior to vitrification. Oocytes were exposed to 1.6 M ethylene glycol (EG) and 1.3M dimethyl sulfoxide (DMSO) in BM and then were exposed to 3.2 M EG, 2.6 M DMSO and 0.5 M sucrose in BM or 3.2 M EG, 2.6 M DMSO and 0.5 M trehalose in BM. Oocytes with cumulus cells and oocytes without cumulus cells were considered as control 1 and control 2, respectively and held in TCM 199-HEPES at $39^{\circ}C$. Oocytes were frozen using modified solid surface vitrification and were stored in cryotubes in liquid nitrogen for more than 1 week. Frozen oocytes were thawed in TCM 199-HEPES containing 0.5 M, 0.25 M and 0.1 M sucrose in BM for 2 min, respectively or 0.5 M, 0.25 M and 0.1 M trehalose in BM for 2 min, respectively. Immunoflurorescence staining of oocytes was performed to assess spindle morphology and chromosome configuration of oocytes. The rates of cleavage and blastocyst were examined following in vitro fertilization. Normal spindle morphology rate of oocytes pre-treated with Taxol prior to vitrification was not higher than that of other vitrified groups. Taxol pre-treatment did not increase cleavage and blastocyst formation rates, although control groups showed significantly higher rates (p<0.05). Percentages of normal spindle and embryonic development were not significantly different among vitrified groups regardless of type of sugar. In conclusion, Taxol pre-treatment of oocytes before cryopreservation did not reduce the damage induced by vitrification and subsequently did not improve embryonic development following vitrification. Trehalose may be used as an alternative non-permeating cryoprotectant in vitrification solution.

• Reproductive and Developmental Biology
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• v.34 no.3
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• pp.185-191
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• 2010

Plasminogen activators (PAs) are serine protease that cleave plasminogen to form the active protease plasmin and may participate in mammalian fertilization. Although correlations have been reported between reactive oxygen species (ROS) and sperm function, the relationship between PA activity and ROS is unknown. We determined the effects of ROS on sperm function and PA activities in boar spermatozoa preincubated under the X-XO system. When spermatozoa were treated with the X+XO group, a significant increase (p<0.05) was observed in the percentage of acrosome reacted spermatozoa compared with that of the control group. However, when antioxidants were added to the medium with X+XO, the rate of acrosome reaction tended to decrease. Also, a significantly lower percentage of acrosome reacted spermatozoa was observed in the X+XO+catalase group at 6 hr of incubation compared with that of X+XO group. The density of malondialdehyde (MDA) was higher in the X+XO group than in different treatment groups. In another experiment, incubation of spermatozoa in medium with X+XO was associated with a significant (p<0.05) increase in activity of tPA-PAI and tPA compared with the control group. Antioxidants decreased the increased activity of tPA-PAI and tPA by preincubation in the X-XO system. Also, a significantly lower (p<0.05) activities of tPA-PAI and tPA were observed in the X+XO+catalase group compared with the X+XO group. No significant differences, however, were observed in the activity of uPA. These results suggest that the increase of acrosome reaction by the X-XO system resulted in increase of PAs activity in the sperm incubation medium.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.2
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• pp.161-167
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• 2001

Objective: Granulocyte-macrophage colony stimulating factors known to be secreted in murine and human reproductive tract. The development of human, bovine and murine embryos could be promoted by addition of GM-CSF in culture medium. However, the pregnancy and implantation rate of embryos cultured in GM-CSF have not been evaluated. The aim of this study was to assess the effect of GM-CSF in embryo development, pregnancy and implantation rate. Methods: A total of 191 IVF cycles were divided into control and GM-CSF supplement group (control=96, GM-CSF=95). The embryos were cultured for three day with or without 2 ng/ml of recombinant human GM-CSF. The quality of embryo, developmental velocity, pregnancy and implantation rates were compared. Results: There was no difference in age, number of gonadotropin ampules used, number of oocytes and fertilization. The number of ICSI cycle was higher in GM-CSF group. In GM-CSF group, G-1 grade embryos were the highest in proportion (56.4%), while G-2 grade embryos were highest (44.3%) in control group. The developmental velocity of embryos were not different between GM-CSF and control group. The pregnancy and implantation rates were significantly higher in GM-CSF group than control (47.4% vs. 33.3%, 17.0% vs. 11.1% respectively). Conclusion: By adding GM-CSF in culture medium, the quality of embryo, pregnancy and implantation rate could be improved.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.185-191
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• 2013

The purpose of this study is to improve production efficiency of vitrified-thawed transgenic bovine embryos. Transgenic bovine embryos were produced by injection of FIV-GFP lentiviral vector into perivitelline space of in vitro matured MII stage oocytes, and then in vitro fertilization. EGFP-expressing transgenic bovine blastocysts were cultured in serum-containing and serum-free medium. These blsatocysts were vitrified by pull and cut (PNC) container made with 0.25 cm plastic straw. Results indicate that total developmental rates of normal IVF embryo cultured in serum-containing and-free medium into blastocyst were not significantly different (22.3 vs 21.5%) and those of GFP-expressing transgenic bovine embryo into blastocyst showed no significant difference between serum-containing (13.9%) and-free medium (13.1%). However, developmental rate of GFP transgenic embryo was significantly (P<0.05) lower than its of normal IVF embryos. In additional study, we vitrified GFP transgenic normal bovine blastocysts using PNC vitrification method. Survival rate of vitrified-thawed GFP transgenic blastocyst (23.1%) was significantly (P<0.05) lower than its of normal blastocysts (68.9%). Although, survival rate of vitrified-thawed GFP transgenic blastocyst was lower than its of normal blastocyst, our result may suggested that PNC vitrification method is feasible to cryopreserve transgenic embryos. Our next plan will be the production of GFP express transgenic bovine derived from vitrified-thawed embryos using PNC method.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.12
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• pp.1820-1826
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• 2007

In this study, we conducted various experiments in order to develop enhanced cultural conditions for in vitro-produced porcine embryos. All embryos were produced by in vitro maturation (IVM) and fertilization (IVF) of immature oocytes from abattoir-derived ovaries. In experiment 1, we cultured IVF embryos in 4 different groups, namely, 0% bovine serum albumin (BSA), 3% BSA, 0.05% Polyvinyl alcohol (PVA), and 0.5% Polyvinylpyrrolidone (PVP) added to the basal fluid cultural medium, Porcine zygote medium 3 (PZM-3). The rates of embryo development were higher in the group where the PZM-3 media had been supplemented with 3% BSA than the other groups. While not statistically significant, the percent of blastocysts and hatched blastocytes were 6.9% and 25.0% in the 3% BSA group vs. 1.2-6.4% and 0-16.7% in the other groups, respectively. In experiment 2, we added 10% fetal bovine serum (FBS) to PZM-3 on day 0 of culture and observed the development rate of blastocysts per day of culture from days 0 to 5. The development rate of blastocysts was higher at 15.6% on day 4 than on any other day, and was significantly higher than on day 0 or day 1 (p<0.05). The development rate of hatched blastocysts was 26.7% on day 4, and was higher than on any other day. In experiment 3, we cultured IVF embryos with different fluid culture media, grouped as 1) PZM-3+0.3% BSA (day0-day7); 2) PZM-3+0.3% BSA${\rightarrow}$day-4) PZM-3+10% FBS; 3) PZM-3+0.3% BSA${\rightarrow}$PZM-3+0.3% BSA+(day-4) FBS 10%; and 4) PZM-3+0.3% BSA+10% FBS (day0-day7). The development rates of blastocysts and hatched blastocysts were 21.5% and 53.1% in group 3, respectively, which was significantly higher than group 4 with respect to blastocyst development (5.2%, p<0.05) but not hatched blastocysts (14.3%). The total cell number (TCN) of blastocysts in group 3 was higher at $37.8{\pm}16.1$ than the other groups at $16.8{\pm}4.4$ - $30.1{\pm}10.9$; however, this was not significantly different. The results of this study showed that PZM-3 containing 0.3% BSA and supplemented with FBS during the later stage of culture on day 4 resulted in better TCNs and an increased rate of hatched blastocysts.