• Weed & Turfgrass Science
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• v.6 no.2
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• pp.124-129
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• 2017

The Experiments were conducted by moderate season culture of each of early, medium and late maturing varieties which were considered to be of strong salt tolerance in low and high salty reclaimed areas (0.2% at the May). This study was carried out to investigate the proper nitrogen fertilizer level and seeding rates at reclaimed saline land in Korea. The proper seeding rates were $40kg\;ha^{-1}$ in 0.2% saline land. The dry matter production of barnyard millet was possibly estimated by exponential functions of $Y=0.0098X^2+0.7030X+2.6267$. Effects of nitrogen rate on agronomic characteristics, forage yield, and chemical composition of barnyard millet to reclaimed tidal land are summarized as follows: The proper nitrogen fertilizer level was $200kg\;ha^{-1}$ in 0.2% saline land. N was absorbed actively before the emergence of the barnyard millet but showed relative decrease thereafter. The early growth of the barnyard millet was inhibited, resulting in the favorable late growth, increased panicle weight and ratio of matured grain. These results suggest that barnyard millet is the most forage crops for cultivation on reclaimed tideland in view of the good emergence and forage production.

• Korean Journal of Organic Agriculture
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• v.6 no.1
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• pp.99-108
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• 1997

The aim of this experiment was to investigate the effect of applying times of cattle slurry on dry matter yield of Orchardgrass, when cattle slurry applied at rates of 30m3(average mineral nitrogen fertilizer equivalent to 120kg) per ㏊ in different dressing times, such as S1(1st and 2nd growth), S2(1st and 3rd growth), S3(1st and 4th growth), S4(2nd and 3rd growth), S5(2nd and 4th growth) and S6(application for 3rd and 4th growth). The results were as follows. 1. The average annual yields of dry matter were produced 6.36~7.42 ton per ㏊ in 1995~1997 when cattle slurry applied at rates of 30m3(average mineral nitrogen fertilizer equivalent to 120kg) per ㏊ in different dressing times, especially these tendency were shown higher annual dry matter in S4 plot9application for 2nd and 3rd growth). Those were higher 2.48~3.54 tons or 1.22~2.28 tons per ㏊ than the yields from non-fertilizing or phosphrous and potassium fertilizing. 2. Relative yields of annual dry matter in cattle slurry application plots were 164(S2 plot)~191(S4 plot)% in comparing with non-fertilizing plot. 3. Dry matter yields according to cutting frequency were highest at the 2nd cut(1.50 ton) and were in the order 3rd cut(0.95 ton)>4th cut(0.75 ton)>1st cut(0.69 ton/㏊). Also, those of cattle slurry application plot per ㏊ were in the order 2nd cut(2.02~2.73 tons)>3rd cut(1.56~2.08 tons)>4th cut(1.07~1.68 tons)>1st cut(1.11~1.45 tons/㏊). 4. Relative yields of annual dry matter in cattle slurry application plots were 164~219, 161~210, 143~212 and 135~182% at the 3rd, 1st, 4th and 2nd cut, respectively in comparing with non-fertilizing plot. 5. The efficiencies of nitrogen on dry matter yield(kg DM/kg N) were 18.1, 21.3 and 34.5kg DM/kg N when cattle slurry applied to Orchardgrass at rates of 30m3 (average mineral fertilizer equivalent to 120 kg) per ㏊ in 1993, 1994 and 1995, respectively, especially these tendency were shown higher efficiencies in S4 plot(application for 2nd 3rd growth). On the other hand, those of the same level fertilization of mineral nitrogen were 43.8, 19.2 and 13.4 kg DM/kg N in 1993, 1994 and 1995, respectively.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.1
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• pp.71-76
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• 1998

This study was carried out to examine whether the developmental capacity of bovine immature oocytes frozen ultra-rapidly using electron microscope (EM) grids and EFS30 can be obtained. As freezing solution, we used EFS30 which consisted of 30% ethylene glycol, 0.5 M sucrose, 18% ficoll and 10% FBS added in D-PBS. As criterior of oocyte viability, the rates of maturation, fertilization and embryonic development were determined. The results obtained in this experiment were summarized as follows: When ultra-rapidly frozen immature oocytes were thawed, 43.2% of them were survived. The rates of maturation (84.1%) and normal 2 pronuclei formation (57.5%) of frozen immature oocytes were not significantly different when compared to those of control (92.5, 65.0%). In addition, the rates of $\geq2$-cell (65.0%) and blastocyst formation (30.8%) of freezing group were not significantly different when compared to those of control (73.7, 35.7%). These results demonstrate that developmental capacity of frozen-thawed bovine immature oocytes can be successfully obtained when survived from the ultra-rapid freezing method using EM grid and EFS30.

• Development and Reproduction
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• v.15 no.4
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• pp.385-391
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• 2011

Human in vitro fertilization and embryo transfer (IVF-ET) program is a general procedure for infertile couples since first successful delivery on 1978 in UK and Korean first on 1985. Recently in Korea, more than 42,000 cases per year of IVF-ET were performed and showed good pregnancy rates compared worldwide data. The human IVF-ET procedure use many consumables, such as ovum pick-up (OPU) needles, centrifuge tubes, culture dish, ICSI pipette, culture media and ET catheters. Major of these materials are supported by the global companies. Thus, Korean IVF-ET program might be placed unstable situation by global economical risks. These uncertain problems could be overcome by the domestic production of IVF-ET materials and consumables. Two times questionnaires for Korean clinicians and researchers about the domestic production were performed and analyzed. Many of them requested domestic OPU needles, ET catheters, culture media and ICSI pipettes under good quality control and quality assurance system. This trial may be contributed to industrialization and to global competence of Korean IVF-ET program. The results of this survey can be provide a fundamental base for development and production of domestic materials and consumables for human IVF-ET program.

• Korean Journal of Soil Science and Fertilizer
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• v.35 no.2
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• pp.105-111
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• 2002

To find out the optimum nitrogen fertilization levels for the leafy vegetables in volcanic ash soils of Cheju island, fertilization effects on chinese cabbage chinese and cabbage were investigated through pot and field experiments. In pot experiment conducted with two volcanic ash soils of Cheju island, optimum rates of nitrogen fertilizer was ranged from 294 to $331kg\;ha^{-1}$ for chinese cabbage. At field experiment with one volcanic soil, the optimum N fertilizer was $331kg\;ha^{-1}$. On the basis of soil organic matters, fertilizer recommendation formula for cabbage, could be established by using 1.03 of comparison factors (F) compared with chinese cabbage : y=344.54-0.285x for chines cabbage, y= 354.88-0.294x for cabbage, where y is the recommendation amount of nitrogen fertilizer with x g $kg^{-1}$ of organic matter in soil. Actual optimum rate of nitrogen fertilizer for chinese cabbage under field condition was much more similar to the value caluculated by the revised nitrogen recommendation formula than the amount of nitrogen fertilizer recommended by the current formula in volcanic ash soil.

• The Korean Journal of Malacology
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• v.29 no.2
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• pp.105-111
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• 2013

The gonadal development of triploid and diploid Pacific abalones, Haliotis discus hannai was histologically investigated in spawning season. Diploid abalones had matured oocytes and spermatozoa, but most triploid had spermatocytes or developing oocytes that was slightly retarded in gonadal development compared to diploid abalones. In spawning experiment of triploid and diploid abalones, spawning rates of diploid male and female were 100%, but those of triploid female was 50% and male was 25% respectively. Investigation of spawned abalone eggs and spermatozoa revealed that length of diploid sperms head were 17.47 ${\mu}m$, breadth of head were 10.31 ${\mu}m$, length of spermatozoa were 130.72 ${\mu}m$, but those of triploid spermatozoa were 11.83 ${\mu}m$, 7.89 ${\mu}m$ and 103.36 ${\mu}m$ respectively. Triploid spermatozoa were significantly small to diploid spermatozoa (p < 0.05). The eggs of diploid and triploid were not different in size. The cross experiment between oocytes produced by triploids and spermatozoa by diploids ($3n{\times}2n$ cross) revealed that no fertilization were occurred, and $2n{\times}3n$ cross also revealed same result.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.10
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• pp.1412-1421
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• 2002

To examine the feasibility of using a sperm vector system for gene transfer, we have investigated the binding and the uptaking of foreign DNA into the sperm nucleus by PCR, in situ hybridization and LSC. We have also examined the transportation of exogenous DNA into oocytes by immunofluorescene via PCR. Sperm cells were incubated with DNA/liposome complexes (1:4 ratio) in fertilization medium with BSA or without BSA. In situ hybridization demonstrated that the transfection rate of sperm cells with and without BSA was 41 and 68% respectively, when the cells were treated with liposome/DNA complexes and 13% for DNA alone. LSC analysis showed that the binding of exogenous DNA was greatly reduced by DNase I treatment which digests DNA bound onto spermatozoa, suggesting that some of the DNA was internalized into the sperm membrane. To find out whether transfected DNA was internalized into sperm intracytomembrane, sperm DNA was amplified by inverse PCR. No PCR products were detected from sperm cells, indicating that the foreign DNA was simply bound onto the sperm membrane. To investigate transfer rates of exogenous DNA into oocytes via sperm cells, we used immunofluorescene method to follow the distribution of foreign DNA via spermatozoa: a few exogenous DNA was located in the cytoplasm of early embryos (13/60, 21.7% for DNA+/liposome+/BSA) and was not located in the pronucleus and/or nucleus. These results suggest that most of the transfected sperm cells could carry the foreign DNA into the egg by in vitro fertilization, but that the transferred DNA is degraded in the developing embryos without stable integration into the zygote genome. Therefore, we have directly injected with transfected sperm cell into oocyte cytoplasm and observed that some of the exogenous DNA was detected in preimplantation embryonic cytoplasm and expressed at preimplantation stages, suggesting that exogenous DNA in early zygote has their integrity. In this study, we have not identified a noble mechanism that interfering transportation of foreign DNA into zygote genome via spermatozoa. Our data, however, demonstrated that inverse PCR and immunofluorescene methods would be used as a new tool for follow-up of gene distribution in oocyte via sperm cells.

• Korean Journal of Ecology and Environment
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• v.53 no.3
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• pp.221-228
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• 2020

We studied the development of the endangered gold-spotted pond frog, Pelophylax chosenicus eggs and analyzed growth and survival rate of the tadpoles while feeding different feed to obtain the basic data on the its biological traits and reproduction. We observed the fertilized eggs of gold-spotted pond frog that naturally spawned in a breeding tank using a microscope. Fertilized eggs were sphere shape surrounded by colloid layers, clearly distinct with animal and plant poles. The fertilized eggs hatched 31 hours after fertilization and operculum complete at 175 hours after fertilization at water temperature of 27.0±1.0℃. As a result of growth and survival rate by different feed from 35 days after mouth open stage, vegetable feed (VF) group and tetra-min (T) group were significantly higher average weight gain rate (WG)(5843.9% and 5736.3%, respectively) and average specific growth rate (SGR) (11.67% and 11.62, respectively) compared with other groups (WG: 641.8~4625.2%, SGR: 5.72~11.01%). The vegetable (V) group showed the lowest growth rate (P<0.05). The average feed efficiency was 177.83% in the VF group, about 17 times higher than the V group. In addition, the average survival rate of the VF and T groups were the high at 97.5% and 100%, respectively, and the V group was the lowest at 32.5%. Therefore, the more efficient feeds for successful breeding is singly with VF or T.

• Korean Journal of Plant Tissue Culture
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• v.22 no.6
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• pp.317-322
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• 1995

As a basic research for breeding new varieties, reciprocal -intergeneric crosses between Citrus junos and P.trifoliata were made. F$_1$ hybrid production using in vitro ovule culture, gametogenesis, and fertilization phenomena were investigated. Frequency of fruit set resulting from crossing of Citrus junos and Poncirus Trifoliata was 16.6% while that of Poncirus Trifoliata and Citrus junos was 11.7%. Callus formation occurred well when ovules at the 6th week after pollination were cultured on MT (Murashige and Tucker) medium supplemented with zeatin 0.5 mg/L and NAA 1.0 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4-D 0.1 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4 D 0.1 or 0.5 mg/L. The invitro germination rates of 20-week-old ovules set C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 54.5% and 48.6%, respectively. The emergence ratios of trifoliate hybrids obtained by C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 56.7% and 100%, respectively. The chromosome number of C. junos and P. Trifoliata was n = 9 or 2n = 18, and the sizes of their pollen grain were 33.75 $\mu$ and 25.0 $\mu$. The length and width of embryo sac in C. junos and P. Trifoliata were 69.38~79.23 $\mu$ and 27.50~38.56 $\mu$, and those of egg cells were 17.50~41.50 $\mu$ and 6.25~8.12$\mu$. Fertilization of C. junos and P. trifoliata terminated 72 h after pollination.

• Journal of Embryo Transfer
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• v.13 no.3
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• pp.227-233
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• 1998

The aim of these experiments was to investigate the effects of oviduct epithelial cells on bovine in vitro fertilization. Oviduct epithelial cell monolayers (OEC) on the 4-well dish were prepared according to general procedures. Monolayers were formed within 5days. The medium for OEC culture (TCM199 with 10% FBS) was replaced with IVF-TALP 2h before each experiment. Macromolecules/proteins from oviductal conditioned medium (OM) were recovered by ultrafiltration, which desalted and concentrated macromolecules greater than 5kDa, and this OM were added to W medium (experiment 1). The cleavage rate in OM+OEC group was significantly higher than in OM group (p〈0.01). In this experiment 2, oocytes were inseminated on OEC with sperm which had been pre-incubated with OEC for 0 or 4h before insemination. In this experiment, oocytes were exposed to sperm only 8 h for clarifying the effect. After insemination, oocytes were cultured in CRlaa. At 42 h post insemination, oocytes were denuded and examined for evidence of cleavage. The cleavage rates of oocytes which were inseminated with OEC treated sperm for 4 h were significantly higher than those of the other group (p〈0.01). In conclusion, sperm released from OEC have more fertilizing ability than those before attachment.