• Clinical and Experimental Reproductive Medicine
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• v.30 no.4
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• pp.333-340
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• 2003

Objective: This study was performed to examine the maturation and the development to the blastocyst stage of immature oocytes collected from patients with high risk of ovarian hyperstimulation syndrome (OHSS). Materials and Methods: Cumulus-oocyte complexes (COCs) were collected following only HCGpriming for non stimulated IVF-ET cycles of the patients. At the time of oocyte collection, COCs were classified into three groups in accordance with their appearance (Group I: oocytes with dispersed cumulus cells; Group II: oocytes with compacted cumulus cells; Group III: oocytes with sparse cumulus cells). The in vitro maturation and blastocyst development rates of the COCs were compared among these groups. From August 2001 to June 2002, 48 IVM/IVF-ET cycles from 42 patients (mean age: $32.4{\pm}3.8$ years) were performed. To prevent the occurrence of OHSS, the patients were primed with 10, 000 IU HCG alone 36 h before oocyte collection without gonadotropin stimulation. Oocytes were aspirated on cycle days from 7 to 13. The normal COCs were classified into three groups according to their appearance. The aspirated immature oocytes were cultured in YS maturation medium containing 30% (v/v) human follicular fluid (HFF), 1 IU/ml FSH, 10 IU/ml HCG and 10 ng/ml rhEGF. Fertilization was induced by intracytoplasmic sperm injection (ICSI). All zygotes were co-cultured with cumulus cells in $10{\mu}l$ YS medium containing 10% HFF until day 7 after oocyte collection. Blastocyst transfer was performed on day 5 after ICSI. Results: Th e mean number of oocytes cultured in the IVM/IVF cycles was $24.7{\pm}10.6$. Of 1185 COCs, those assigned to Group I, II and III were 470 (39.7%), 414 (35.0%) and 301 (25.4%), respectively. The maturation rate (94.5%, 444/470, p<0.05) in Group I was significantly higher than those of Group II (62.8%, 260/414) and Group III (73.1%, 220/301). Especially, 30.9% of COCs in Group I (145/470) was matured on the day of oocyte aspiration. There were no differences in the rates of fertilization and cleavage among the three groups. The development rate to the blastocyst stage in Group I (54.6%, 206/377, p<0.05) was also significantly higher than those in Group II (33.0%, 68/206) and Group III (30.1%, 52/173). Twenty-four clinical pregnancies (50.0%) was obtained and 22 pregnancies (45.8%) are ongoing. Implantation rate in the present study was 24.6%. Conclusion: These results suggest that there is a positive correlation between the appearance of COCs and the developmental competence of the immature oocytes in non stimulated IVM/IVF cycles.

• Weed & Turfgrass Science
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• v.3 no.4
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• pp.342-346
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• 2014

The purpose of this experiment is for investigating how much the amino acid liquid fertilizer and composted liquid manure, culture microorganism effect on the breeding of grasses and knowing the extent of the thatch content through an analysis of the soil. For testing about soil chemical, the quality of grasses, and the extent of the thatch content in the soil, we cultivated 6 kinds of microorganisms having the effect of thatch dissolution and sprayed these 6 microorganisms with composted liquid manure and the amino acid liquid fertilizer on the place Creeping bentgrass have planted. This conduction started from June to October, 2012 and 2013 (The interval: 2 weeks). In the result of the turf growth, there is no big difference between soil chemical and trace element. And we can know Leaf Color Index, Chlorophyll Index and Root Length are almost same as among treatment. In conclusion, the mixing fertilization of culture microorganism and composted liquid manure is better effective than the traditional fertilization. And it can be expected the effect of the quality of grass and Thatch decomposition in soil.

• Horticultural Science & Technology
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• v.35 no.4
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• pp.431-438
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• 2017

This study investigated the effect of pre-planting nitrogen (N) fertilization levels added to a soilless root medium on the growth of 'Dotaerang Dia' tomato seedlings. The N levels were varied for a total of 7 treatments: 0, 100, 250, 500, 750, 1,000, or $1,500mg{\cdot}L^{-1}$. The pH of the root media gradually rose in all treatments as the seedlings grew; however, the differences in the pH were not significant among the treatments. The electrical conductivity (EC) of the root media was significantly different among the treatments from sowing to week three, then drastically decreased after week four, which diminished the differences in the EC among the treatments. At week six, plant height, leaf length, leaf width, number of leaves, and fresh and dry weights of the shoot were highest for the treatment with $500mg{\cdot}L^{-1}N$. In contrast, the treatment with $1,500mg{\cdot}L^{-1}N$ had the lowest results for all growth measurements. The fresh weight was 67% heavier in the $500mg{\cdot}L^{-1}N$ treatment compared to the $1,500mg{\cdot}L^{-1}N$ treatment. The total N content in the tissues was lowest in the treatment with $0mg{\cdot}L^{-1}N$ and highest in the treatment with $1,000mg{\cdot}L^{-1}N$. The contents of calcium (Ca), magnesium (Mg), and metal micronutrients in the tissues were highest in the $250mg{\cdot}L^{-1}N$ treatment. A previous study demonstrated that adjusting the fertilization level to promote growth to over 90% of the maximum growth is a good strategy for lowering production costs and preventing damage due to excessive fertilizer absorption by crops. Our results indicated that the optimal pre-planting N fertilization level for tomato plug seedlings should be lower than $500mg{\cdot}L^{-1}$ and the optimum tissue N contents should be around 3.21% to 4.60%.

• Korean Journal of Plant Tissue Culture
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• v.22 no.5
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• pp.261-266
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• 1995

This study was carried out to develop new varieties which are dwarf and tolerant to winter cold in the Aurantioideae by intergeneric crossing. to do that, the reciprocal crosses of Hwanggeumyooza and trifoliate orange, yooza and trifoliate orange were done and in vitro immature ovule culture of their hybrid was carried out .The callus formation from immature ovule was good in order of Hwanggeumyooza, Hwanggeumyooza $\times$ tifoliate orange, yooza, and trifoliate orange and best at 1 to 3 mg/L NAA+0.5mg/L zeatin on MT medium. In vitro germination percentage of 20week old hybrid of Hwanggeumyooza $\times$ tifoliate orange and trifoliate orange $\times$ Hwanggeumyooza were 41.3% and 37.7, respectively. The phenotype of hybrid (95%) of Hwanggeumyooza $\times$ trifoliate orange and that (100%) of trifoliate orange $\times$ Hwanggeumyooza were similar to that of trifoliate orange. After Hwanggeumyooza was pollinated by pollens of trifoliate orange, the pollen tubes grew on stigma after 3h of pollination and entered into micropyle after about 24~28 h. One gamete in pollen was fused with polar nuclei after 2 days and other one fused with egg nucleus at 3days after pollination. The fruit set percentage by intergeneric crossing was 14.0% in Hwanggeumyooza $\times$ trtfoliate orange and 17.5% in trifoliate orange $\times$ Hwanggeumyooza. The fruit set percentages of Hwanggeumyooza. and trifoliate orange were 34.2% and 39.5% by artificial self-fertilization, 34.2% and 39.5% by artificial cross fertilization, 3.1% and 1.4% by parthenocarpy and 13.0% and 3.0% by natural fertilization, respectively. The somatic and gametic chromosome numbers of Hwanggeumyooza, yooza, and trifoliate orange were 2n=18 and n=9.

• Korean Journal of Animal Reproduction
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• v.20 no.2
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• pp.127-134
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• 1996

The objective of this experiment was to test the ability of the fertilization of EGF treated pig oocytes for in vitro maturation. The addition of EGF (10 ng/ml), FSH (10 ${\mu}\textrm{g}$/ml), or FBS (10%) on maturation medium of pig immature oocytes divided into four groups as follows; group 1: untreatment, group 2: EGF alone, group 3: combination of FSH and FBS, or group 4: combination of EGF, FSH, and FBS. The interactive effects of nuclear maturation rates (M II%) of EGF alone, FSH plus FBS, and EGF plus FSH added FBS treatments were significantly higher than those of non-treatments (P<0.001). The fertilization rate of EGF alone (group 2) was lower than that of 3, 4 groups, but was significantly higher than group 1 (p< 0.005). Furthermore, combination of EGF, FSH,and FBS (group 4) was higher than others (group 1. 2, 3) on male pronuclei formation as well as penetration of sperm (P<0.05). These results suggested that EGF alone decreased the ability of cytoplasmic maturation compared to nuclear maturation in pig oocytes, but a high level of cytoplasmic maturation of in vitro-matured pig oocytes can be achieved when supplemented with FSH and FBS.

• Journal of Embryo Transfer
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• v.10 no.2
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• pp.121-129
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• 1995

To examine the critical effect of oxygen concentration on embryonic development, in vitro fertilized embryos were cultured in media(TCM199 vs. SOF) supplemented sera(1O% FCS vs. 10% HS) with and without bovine oviduct epithelial cells under two gas atmosphere (5% $CO_2$ in air vs. 5% $CO_2$, 5% $O_2$, 90% $N_2$). Oocytes, obtained from abattoir ovaries, were matured in EGF containing TCM199 medium co-cultured with BOEC for 24 hours, followed by exposure to frozen-thawed, heparin4reated spermatozoa in TALP for 30 hours. And then early embryos(1~2 cell) were cultured in both TCM199 and SOF supplemented with 10% FCS or 10% RS under 5% $CO_2$ in air or 5% COi, 5% $O_2$, 90% $N_2$. Development to morulae and blastocysts was recorded on days 7, after the start of in vitro fertilization. The developmental rates of in vitro fertilized embryos to morulae and blastocysts cultured in SOF with BOEC under 5% $CO_2$, 5% $O_2$, 90% $N_2$(24.4%) were significantly(p<0.05) higher than cultured in SOF with BOEC under 5% $CO_2$ in air(14.1%) at seven days after in vitro fertilization. When early bovine embryos were cultured in TCM 199 and SOF under two different gas atmosphere, there were no significant differences in the developmental rates to morulae and blastocysts between supplements of 10% FCS and 10% HS. The rates of development to morulae and blastocysts were significantly(p<0.01) higher in TCM 199 with BOEC(24.7%) than TCM199 without BOEC(10.9%) under 5% $CO_2$ in air, otherwise SOF without BOEC(36.4%) were significantly (p<0.05) higher than in SOF with BOEC (24.4%) under 5% $CO_2$, 5% $O_2$, 90% $N_2$. In summary, these experiments have proved that the culture system in SOF supplemented 10% ES is effective on in vitro development of early bovine embryos under 5% $CO_2$, 5% $O_2$, 90% $N_2$. In addition, it is effective to development of bovine embryos that TCM 199 should be co-cultured with BOEC and SOF should be cultured without somatic cells under two different gas atmosphere.

• Korean Journal of Plant Resources
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• v.25 no.2
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• pp.285-292
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• 2012

Self-incompatibility (SI) system is a genetic barrier that prevents self-fertilization and promotes cross-pollination among different S genotypes. In many of these species, SI is controlled by a single genetic locus known as S locus, which prevents the fertilization by pollen with same locus. S RNases are the products of the S-locus expressed in the stylar tissue of Fuji Apple with gametophytic self-incompatibility system. This study investigated the various types of chemicals in order to select more effective inhibitors and activators. The effect on the inhibition of S RNase of Fuji apples was investigated $in$ $vitro$. The result showed that the enzyme activity was reduced 24.3% by Iron(II) Sulfate, significantly. $In$ $vitro$ studies of pollen growth tube showed that pollen tube growth had a higher germination rate (90%) in 10% Sucrose than in 2% sucrose extension medium. Data on the fruit set of apples treated with inhibitor and activator. Double application of $A^+$(Apple Plus, ISTECH Co. Ltd.,)+Vitamin B6 had the highest central fruit set as 86.1%(Andong). One time application of $A^{++}$Vitamin B1 in Yeongju obtained the highest central fruit set (91.9%).

• FLOWER RESEARCH JOURNAL
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• v.19 no.4
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• pp.231-237
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• 2011

In this study, we performed ovule culture after reciprocal crosses of two Alstroemeria accessions and investigated genetic contribution of parents by using RAPD markers. The best method was half-ovule culture on MS medium supplemented with $60g{\cdot}L^{-1}$ sucrose and $2.2g{\cdot}L^{-1}$ gelrite at 14 days after pollination. Embryos began to germinate after 6 weeks of culture. The complete plantlets were formed after 4 months of culture. In eight progenies and two parental cultivars, 59 polymorphic bands were obtained out of 89 total bands by RAPD analysis using 7 primers. Eight $F_1$ progenies from the crosses between two accessions using reciprocal crosses showed 1:1 contribution of maternal and paternal parents. It is confirmed that $F_1$ progenies were obtained from parental accessions by using RAPD markers. We conclude this cross combination showed pre-fertilization barriers with incompatibility between stigma or style, and pollen because progeny number was different in each cross combination. Thereby, it warrants overcoming pre-fertilization barrier together with post-fertilization barrier in order to broaden the heterozygosity within progeny populations in Alstroemeria breeding program.

• Journal of Veterinary Science
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• v.24 no.1
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• pp.4.1-4.16
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• 2023

Background: In vitro culture of preantral follicles is a promising technology for fertility preservation. Objectives: This study aims to investigate an optimized three-dimensional (3D) fetal bovine serum (FBS)-free preantral follicle culture system having a simple and easy operation. Methods: The isolated follicles from mouse ovaries were randomly divided in an ultra-low attachment 96-well plates supplement with FBS or bovine serum albumin (BSA) culture or encapsulated with an alginate supplement with FBS or BSA culture. Meanwhile, estradiol (E₂) concentration was assessed through enzyme-linked immunosorbent assay of culture supernatants. The diameter of follicular growth was measured, and the lumen of the follicle was photographed. Spindle microtubules of oocytes were detected via immunofluorescence. The ability of oocytes to fertilize was assessed using in vitro fertilization. Results: The diameters were larger for the growing secondary follicles cultured in ultra-low attachment 96-well plates than in the alginate gel on days 6, 8, and 10 (p < 0.05). Meanwhile, the E2 concentration in the BSA-supplemented medium was significantly higher in the alginate gel than in the other three groups on days 6 and 8 (p < 0.05), and the oocytes in the FBS-free system could complete meiosis and fertilization in vitro. Conclusions: The present study furnishes insights into the mature oocytes obtained from the 3D culture of the preantral follicle by using ultra-low attachment 96-well plate with an FBS-free system in vitro and supports the clinical practices to achieve competent, mature oocytes for in vitro fertilization.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.163-172
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• 2001

The present study was carried out to examine the effects of $O_2$concentrations and culture media on in vitro maturation and embryo development of porcine follicular oocytes. The results were summarized as fellows : 1. The rates of GVBD and nuclear maturation in NCSU-23 and TCM-199 media with 10% PFF under the conditions of 5% and/or 20% $O_2$concentrations were not different among the each treatment groups(P>0.05). 2. The rates of polyspermy and mean numbers of penetrated sperm were significantly lower in NCSU-23 medium than in TCM-199 medium (P>0.05). However, the rates of polyspermy and mean numbers of penetrated sperm were not different between 5% and 20% $O_2$concentrations. 3. The rates of blastocyst formation at day 7 after in vitro fertilization were significantly higher in NCSU-23 medium under the condition of 20% $O_2$concentration than in TCM-l99 medium under the condition of 5% or 20% $O_2$concentrations. However, the rates of blastocyst formation and total cell numbers in blastocysts were not different between 5% and 20% $O_2$concentrations. In conclusion, the use of NCSU-23 medium under the condition of 20% $O_2$concentration was beneficial for porcine oocyte maturation and in vitro development.