• Journal of Embryo Transfer
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• v.28 no.3
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• pp.169-175
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• 2013

Recently, the transgenic animal production technique is very important for the production of bio-parmaceutical as animal bio-reactor system. However, the absence of survival evaluation in vitro produced transgenic embryos has been a problem of the low productivity of transgenic animal because of absent of pre-estimate of pregnancy after transgenic embryos transferred into recipient. Therefore, this study is conducted to improve efficiency of transgenic cattle production by improving the non-surgical embryo transfer (ET) method. Transgenic bovine embryos were produced by injection of feline immunodeficiency virus enhanced green fluorescent protein (FIV-EGFP) lentiviral vector into perivitelline space of in vitro matured MII stage oocytes, and then in vitro fertilization (IVF) was occured. Normal IVF and EGFP expressing blastocysts were transferred into recipients. Results indicated that 2 expanded blastocysts (34.7%) transferred group showed significantly (P<0.05) higher pregnancy rate than 1 expanded blastocyst (26.8%) transferred group. In case of parity of recipient, ET to heifer (34.9%) showed significantly (P<0.05) higher pregnancy rate than ET to multiparous recipient (21.2%). However, there are no significant differences of pregnancy rate between natural induced estrus and artificial induced estrus groups. Significantly (P<0.05) higher pregnancy rate was obtained from recipient group which have normal corpus luteum with crown group (34.8%) than normal corpus luteum without crown (13.6%). Additionally, treatment of $100{\mu}g$ Gn-RH injection to recipient group (38.6%) 1 day before ET significantly (P<0.05) increase pregnancy rate than non- Gn-RH injection to recipient group (38.6%). We also transferred 2 EGFP expressing expanded blastocysts to each 19 recipients, 7 recipients were pregnant and finally 5 EGFP transgenic cattle were produced under described ET condition. Therefore, our result suggested that transfer of 2 good-quality expanded blastocysts to $100{\mu}g$ of Gn-RH injected recipient which have normal corpus luteum with crown is feasible to produce transgenic cattle.

• 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.

• Journal of Aquaculture
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• v.18 no.2
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• pp.115-121
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• 2005

This study was performed to investigate the formation and mass production of resting eggs by freshwater rotifer, Brachionus calyciflorus as influenced by different aeration supplements and exchange intervals of culture water in 15-L culture vessels and 500 L culture tanks. The maximum densities and mixis rates of the rotifers were not different between experimental group exposed to air or oxygen supplements. However, the fertilization rate and formation of resting eggs of the rotifers in the air-supplemented group were significantly higher than those in the oxygen supplemented group. In the experiment concerning exchange interval of culture water, the maximum density of the rotifers and formation of resting eggs in the batch culture were significantly higher than those in the semi-continuous culture with exchange of water every day. The formation of resting eggs per Chlorella dry weight was highest in the semi-continuous culture with exchange of water every day. The resting eggs of rotifers were produced at a density of $51.8{\sim}57.9{\times}10^6$ eggs in 500-L culture tanks. In this study, the batch culture with air is an effective method for mass production of resting eggs by the freshwater rotifer, B. calyciflorus, and the efficiency of mass production of resting eggs by this rotifer was similar to that of the seawater rotifers, B. plicatilis and B. rotundiformis.

• Journal of Embryo Transfer
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• v.9 no.1
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• pp.95-102
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• 1994

본 연구는 종모우의 선발방법으로 난포란을 이용하여 실험실내 정자의 수정능력을 직접 검정하여 평가코자 시도되었다. 즉 본 실험은 후대검정중에 있는 한우 후보종모우 15두의 동결융해정자의 수정능력을 평가하기 위하여 정액을 고장액(HIS)에 처리한 후 DM에서 6시간 그리고 소 난포액이 20% 첨가된 DM에서 4시간 전배양하여 수정능을 획득시켜 정자의 활력과 첨체 반응율을 조사하였고 전배양된 정자의 체내(토끼 난관) 또는 체외수정능력을 조사하기 위하여 FCS 15%, 발정암소혈정(CSS) 10%가 첨가된 mKRB에서 체외성숙된 한우난포란과 수정시켜 수정능력을 평가하였으며 인공수정에 의한 개체별 수태율과도 비교 검토한 바 다음과 같은 결론을 얻었다. 1. 한우 난포란의 체외성숙율은 BSA 첨가구 에서 43.8%, FCS 15% 첨가구에서 67.4%, CSS10% 첨가구에서 69.9%이었다. 2. 토끼 난관에서 체외수정율은 BSA 참가구에서 43.8%, FCS 15% 참가구 41.2% 및 CSS 10% 참가구 35.0% 이었다. 3. 후보종무우 15두의 정액을 HIS-DM으로 처리후 6시간 전배양하였을 때 정자의 활력지수는 9-32%였고 첨체반응율은 19-44% 이었으며 20% 난포액을 첨가하여 4시간 전배양 하였을 때 정자의 활력지수는 9-13% 이었고 첨체반응율은 20-43%로 개체간에 차이가 있었다. 4. 체외수정율은 6.6-85.7%였으며, 발정암소혈청(CSS) 10%가 첨가된 mKRB에서 성숙시킨 난포란이 FCS 15% 첨가된 mKRB에서 성숙시킨 난포란보다 다소 높았으나, 정자수정능획득방법간에는 차이가 없었 다. 5. 체외수정율에 있어서 전배양후 정자활력지수와는 부의 상관이 었으며, 첨체반응율과는 낮은 정의 상 관을 나타냈다. 6. 종모우의 수태율은 체외수정율, 정자활력지수 및 첨체반응율과 낮은 정의 상관관계를 나타냈다. 7. 종모우의 개체간 수태율 우열순위에서는 수정율순위와의 사이에 더욱 낮은 부의 상관관계를 보였다. 8. 이상의 연구결과 비록 후대정검중의 제한된 자료로 인하여 종모우 수태율과 체외수정율간에 유의적 인 상관관계는 없었으나, 연결 한우 수정율 평가에 대한 실험실내의 검정가능성을 찾을 수 있었다.

• FLOWER RESEARCH JOURNAL
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• v.18 no.3
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• pp.193-200
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• 2010

Current rose cultivars are all composed of heterozygous genome due to long history of out crossing including interspecific hybridization. It has been adapted by artificial selection and crossing by breeders that mainly based on the crossing with fertile pollen derived from inter- or intra-specific hybridization. Pollen viability and germination ability tests provide valuable information for the designing of parentage for more successful breeding efficacy. In this study, we tested the pollen viability and germination ability in seven rose cultivars to find any relationship among several factors including pollen size, ploidy levels, and crossing compatibility. The pollen viability showed wide ranges from 39% 'Pinocchio' as minimum to 82% 'Scarlet Mimi' as maximum, whereas pollen germination rate were from 1% 'Mini Rosa' to 41% 'Scarlet Mimi' as a highest. Pollen size ranged from 41.3 to $45.4{\mu}m$ in large sized pollen and 30.7 to $37.4{\mu}m$ in small sized pollen. The mean diameter of large sized pollen is approximately 10-40% bigger than that of small sized pollen. There are positive relationships among ploidy level, total chromosome length, and pollen size. Crossing list showed that seed setting ratio and seed germination were related to pollen viability, pollen germination, and ploidy level.

• Journal of the Korea Organic Resources Recycling Association
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• v.27 no.1
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• pp.5-14
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• 2019

The objective of this study was investigated the effect of organic fertilizer(OF) application on the grain yield, protein content, and soil chemical properties in rice cultivation to evaluate the replacement ratio of OF based on basal fertilization of inorganic(nitrogen) fertilizer, In case of soil 1 with optimum range of organic matter for rice growth, the grain yield of rice in 70% and 100% treatments of OF were higher than the other treatments. In soil 2 with higher than the optimum range of soil organic matter, the grain yield of rice of NPK, 30%, 70%, 100%, and 300% treatments of OF showed no significant difference. When the protein criteria of rice quality was below 7%, the treatment of 30%, 70%, and 100% using OF in soil 1 were satisfied with protein criteria of rice. In case of soil 1, ammonium nitrogen content, electrical conductivity, and the organic matter in 30%, 70%, 100%, 300% treatments of OF showed similar tendency to NPK treatment. These results suggested that the substitution ratio of OF 70~100% could be appropriate to increase the yield and commercial quality for rice.

• Journal of Korean Society of Forest Science
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• v.110 no.2
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• pp.189-197
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• 2021

This study aims to optimize the appropriate concentration of fertilizers for Sageretia thea by analyzing growth performances (height and root collar diameter) and physiological characteristics (photosynthesis, chlorophyll contents, and chlorophyll fluorescence reaction). As fertilizer concentration was increased to 1.5 g·L^-1, growth increased, but it decreased at 2.0 g·L^-1 treatment. Root collar diameter growth was reduced because of higher fertilizer concentrations. Photosynthesis reactions showed the highest CO₂ reaction curves, maximum photosynthesis rate, and maximum carboxylation rate in the 1.5 g·L^-1 fertilizer treatment. The chlorophyll fluorescence reaction and SPAD values revealed that fertilizer treatment improves photosynthesis efficiency and robustness compared with untreated control. Therefore, the appropriate fertilizer concentration for producing good seedling quality of Sageretia thea is 1.0~1.5 g·L^-1.

• Korean Journal of Veterinary Service
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• v.44 no.4
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• pp.227-237
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• 2021

Artificial insemination of Korean native cattle (KNC) is the predominant method for breed improvement. However, industrialization of embryo production and transfer is necessary to utilize the genetic potential of KNC. The aim of this study was to examine associations between KNC donor cows and ovum pick-up (OPU) conditions, in-vivo oocyte recovery, and embryo development. Oocyte recovery and blastocyst development rates were higher at 50 and 60 mmHg OPU vacuum pressure than at 40 mmHg, which was, however, not significant. Regarding follicle growth, injection of 500 ㎍ GnRH 36 hours before OPU significantly increased the number of OPU oocytes from an average of 4.6 to 7.6 (P<0.05); no significant difference in embryo development rates was observed. Significant differences were observed in the numbers of OPU oocytes, embryo development rates, and transplantable blastocysts per individual among nine KNC donors (P<0.05). Furthermore, although there was no difference in OPU oocyte recovery intervals in approximately 2~8 weeks, the number of recovered oocytes significantly decreased at the 12-week interval (P<0.05); there was no difference in embryo development rates. The number of oocytes and embryonic development rates only tended to decrease until the seventh OPU session, but decreased significantly until the eighth session (P<0.05). The average pregnancy rate after transfer of OPU-derived in-vitro embryos into recipient cows was 41.8%. To improve the efficiency of OPU egg recovery and in-vitro embryo production, considering KNC donor characteristics, vacuum pressure of 60 mmHg, GnRH pretreatment to induce follicle growth, and effective OPU egg recovery up to seven times at intervals of 2~4 weeks appears to be most suitable. This study may facilitate the industrialization of KNC embryo production and transfer using high-quality cows.

• Korean Journal of Soil Science and Fertilizer
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• v.35 no.5
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• pp.280-289
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• 2002

This study was conducted to evaluate the effects of annual application of slowle cooled slag silicate fertilizer on rice yield and soil chemical properties. Field experiment was done on the condition of fertilization of silicate fertilizer 1,500 and $2,500kg\;ha^{-1}$ to the clay loam paddy field during 26 years from 1975 to 2000. The results obtained were as follows; Rice yield of NPK+silicate fertilizer $1,500kg\;ha^{-1}$ and $2,500kg\;ha^{-1}$ were increased by 15%, 8% respectively incomparion with NPK control plot in 2000($26^{th}$ year). Changes in average rice yield for 5 years interval were continually showed increase in the treat of silicate fertilizer $1,500kg\;ha^{-1}$ compared to NPK and NPK+silicate fertilizer $2,500kg\;ha^{-1}$ treated plot. The amounts of N, $K_2O$, CaO and MgO in the treat of silicate fertilizer $1,500kg\;ha^{-1}$ were much more than those of silicate fertilizer 2,500kg treated plot, and the treat of silicate fertilizer $1,500kg\;ha^{-1}$ showed higher in nutrients availability and fertilizers use efficiency than other treated plots at harvesting stage. Amount of N, $P_2O_5$, $K_2O$ and MgO in unhulled rice grain, those of CaO, MgO and $K_2O$ in rice straw and those of $K_2O$ and $SiO_2$ in rice root were positively recognized significant relationships with grain yield. According to soil analysis after experiment in 2000, the silicate fertilizer $2,500kg\;ha^{-1}$ annually applied plot were increased especially in soil organic matter, CEC and available phosphate content in comparison with NPK+silicate fertilizer $2,500kg\;ha^{-1}$ applied plot.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.153-163
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• 2002

In vitro matured porcine oocytes have very small volume of perivitellinspace (PVS). In these respect, the effect of sucrose and polybrene on the efficiency of gene transfer was investigated. As a gene (hGH) transfer vehicle, vesicular stomatitis virus glycoprotein pseudotyped retroviral vector (VSV-G) was used. Sucrose treatment has no detrimental effect on the rates of cleavage and resulted in the enlargement of PVS for the efficient introduction of retroviral vector stocks. Introduction rates of retrovirus in 0.5, 1, 2, 3 % sucrose treatment group were higher than that of the non-treatment group (39.3, 43.3, 35.7, 40.7 % vs. 8.3 %), respectively. In addition, we observed that sucrose pretreatment during injection procedure significantly reduce the frequency of polyspermy. In general, polybrene is a polycation essential for retrovirus transduction. The groups with the addition of 0.5, 5, 50$\mu\textrm{g}$/$m\ell$ polybrene exhibited a significant effect on gene transfer compared to that of the non-addition group (56.5, 50.0, 57.1 % vs. 34.6 %), respectively But, when the oocytes were co-injected with retrovirus and 50$\mu\textrm{g}$/$m\ell$ polybrene, the rates of cleavage and blastocyst development were 43.3 and 4.6%, respectively. This rates were lower than those of the non-addition group (70.0 and 17.3 %). In conclusion, sucrose pretreatment have increased efficiency of retroviral mediated gene transfer in porcine oocytes with no damage on in vitro fertilization and embryo development. In addition, sucrose pretreatment was beneficial in polyspermy inhibition. Presence of polybrene during microinjection showed a beneficial effect on the gene transfer in porcine oocytes, in low concentration. And these results will provide an useful tool for production of transgenic pigs by retroviral mediated gene transfer.