• Korean Journal of Plant Resources
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• v.31 no.5
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• pp.515-521
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• 2018

This was carried out to develop a chromosome-doubled (12x) persimmon that will be used as a crossing parent to select seedless persimmon cultivars with the change of the consumption trend recently. To obtain a chromosome-doubled (12x) persimmon, colchicine was applied at the meristem of seedlings in vitro derived from cross among hexaploid persimmon (Diopyros kaki Thunb.). These were treated with 0.03%, 0.05% and 0.1% colchicine respectively for doubling chromosome, and it was most effective at the concentration of 0.05% colchicine. After colchicine treatment, we conducted tests to elucidate conditions for inducing shoot and root development. As the result, the shoots grew best when cultivated at 1/2MS media plus 10 and $30{\mu}M$ zeatin respectively, and the roots grew best when cultivated at 1/2MS media after dipping for 5 seconds at 10 mM NAA+5% DMSO. We also compared seedlings that have chromosome (6x) do not doubled and crossing parents (6x) and chromosome-doubled seedlings (12x). As the result, these chromosome-doubled seedlings (12x) showed lower stomatal density and larger stomatal size.

• Journal of Environmental Impact Assessment
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• v.22 no.6
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• pp.745-751
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• 2013

Environmental Specimen Banks (ESBs) have been established widely in the world since 1979 for monitoring long-term bio-accumulation of environmental pollutants. The ESBs perform the retrospective analysis of representative biological samples collected regularly and store them in cryogenic condition. In Korea, National Environmental Specimen Bank (NESB) was established in 2009. Since then, NESB had prepared the standard operating procedures (SOPs) for the seven kinds of specimens (Red Pine (Pinus densiflora), Common Carp (Cyprinus carpio), and etc.) for monitoring the effect of environmental pollution on the terrestrial ecosystem and river ecosystem. In 2012, NESB added Black-tailed Gull (Larus crassirostris)'s eggs to the list of the environmental specimen for monitoring marine environmental pollution. In this study, we sampled the eggs of the Black-tailed Gull on two islands (Baekryeongdo of the West Sea and Hongdo of the South Sea). Especially, we selected eggs which remained in the early stages of embryo development for certifying the consistent and stable monitoring for environmental pollutants in egg contents. However, it was not considered to classify an order of eggs per clutch in this study. It is known that the concentration variations of pollutants exist among eggs in one clutch. Therefore, it is needed to consider the positions of eggs in the laying sequences to meet the objective of bioaccumulation monitoring. We collected 30 eggs in each site but the amount may be insufficient to achieve storage target (over 2,000 g). Therefore, we need to consider an optimal sampling size.

• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.281-291
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• 2003

In this review, we described the catalogues of the rice proteome which were constructed in our program, and functional characterization of some of these proteins was discussed. Mass-spectrometry is the most prevalent technique to rapidly identify a large number of proteome analysis. However, the conventional Western blotting/sequencing technique has been used in many laboratories. As a first step to efficiently construct protein cata-file in proteome analysis of major cereals, we have analyzed the N-terminal sequences of 100 rice embryo proteins and 70 wheat spike proteins separated by two-dimensional electrophoresis. Edman degradation revealed the N-terminal peptide sequences of only 31 rice proteins and 47 wheat proteins, suggesting that the rest of separated protein sports are N-terminally blocked. To efficiently determine the internal sequence of blocked proteins, we have developed a modified Cleveland peptide mapping method. Using this above method, the internal sequences of all blocked rice proteins(i, e., 69 proteins) were determined. Among these 100 rice proteins, thirty were proteins for which homologous sequence in the rice genome database could be identified. However, the rest of the proteins lacked homologous proteins. This appears to be consistent with the fact that about 45% of total rice cDNA have been deposited in the EMBL database. Also, the major proteins involved in the growth and development of rice can be identified using the proteome approach. Some of these proteins, including a calcium-binding protein that tuned out to be calreticulin, gibberellin-binding protein, which is ribulose-1.5-bisphosphate carboxylase/oxygense active in rice, and leginsulin-binding protein in soybean have functions in the signal transduction pathway. Proteomics is well suited not only to determine interaction between pairs of proteins, but also to identify multisubunit complexes. Currently, a protein-protein interaction database for plant proteins(http://genome.c.kanazawa-u.ac.jp/Y2H)could be a very useful tool for the plant research community. Also, the information thus obtained from the plant proteome would be helpful in predicting the function of the unknown proteins and would be useful be in the plant molecular breeding.

• Korean Journal of Plant Tissue Culture
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• v.24 no.4
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• pp.239-256
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• 1997

Plants belonging to the category of 2n apomixis or agamospermy form embryos and seeds without the processes of normal meiosis and syngamy. Seeds produced in this way have identical genotype of their maternal parent. Three different types of agamospermy are recognized: diplospory, apospory, and adventitious (adventive) embryony. $F_1$ hybrid cultivars cannot be used as seed sources in the next ($F_2$) generation because this generation would be extremely variable as a result of genetic segregation. Hybrid vigor is also reduced in the $F_2$ generation. Therefore, parental stocks for hybrid seed production need to be maintained and cross must be continuously repeated. Agamospermic 2n apomixis would make it possible to fix the genotype of a superior variety so that clonal seeds faithfully representing that genotype could be continuously and cheaply produced independent of pollination. That is, $F_1$ hybrid seeds could be produced for many generations without loss of vigor or genotype alteration. Production of apomictic $F_1$ hybrid seed would be simplified because line isolation would not be necessary to produce seed or to maintain parental lines, and the use of male-sterile lines could be avoided. Overall, apomixis would enable a significant reduction in hybrid seed production costs. Additionally, the production of clonal seed is not only important for seed propagated crops, but also for the propagation of heterozygous fruit trees and timbers. Clonal seed would help avoid costly and time-consuming vegetative propagating methods that are currently used to ensure the large-scale production of these plants. Apomixis is scattered throughout the plant kingdom, but few important agricultural crops possess this trait Therefore, most research to date has centered on introgressing the trait of apomixis into agricultural crops such as wheat, maize, and some forage grasses from wild distant relatives by traditional cross breeding. The classical breeding approach, however is slow and often impeded by many breeding barriers. These problems could be surmounted by taking mutagenesis or molecular approach. Arabidopsis thaliana is a tiny sexually reproducing plant and is convenient in constructing and screening in molecular researches. Male-sterile mutants of Arabidopsis are particularly suitable genetic background for mutagenesis and screening for apomictic mutants. Molecular approaches towards isolating the genes controlling the apomictic process are feasible. Direct isolation of genes conferring apomixis development would greatly facilitate the transfer of this trait to wide variety of crops. Such studies are now in progress.

• Korean Journal of Ichthyology
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• v.10 no.2
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• pp.207-215
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• 1998

Just after hatching the embryo has a yolk sac and straighted digestive tract. Just after parturition in 5.0mm TL the mouth and anus were opened. In the larval stage of 5.2mm TL, digestive tract could be divided into pharynx, esophagus, stomach, pyloric caeca and intestine of the early differentiated state. The esophagus of larvae in 5.3mm TL has a PAS positive mucous cell. From over 5.7mm TL, PAS positive goblet cell recognized in the intestine. PAS positive absorptive cell began to appear in the intestine from 5.9mm TL. Yolky materials were absorbed completely in 6.0mm TL. In the larvae of B.5mm TL, gastric glands were observed in the mucosa, but the surface epithelium did not have PAS positive granules. From over 9.0mm TL, the histological structure of esophagus showed similar to adult. In the juvenile stage from 13.0mm to 15.0mm TL, histological structure of the stomach, pyloric caeca and intestine showed similar to adult. From the ultrastructural and histochemical study, it is concluded that the functional digestive tract is present in the juvenile stage from 18.0mm to 20.0mm TL.

• Journal of Forest and Environmental Science
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• v.11 no.1
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• pp.81-101
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• 1995

Random amplified polymorphic DNA (RAPD) technology, a recent approach in molecular genetics, is much usable to select the elite trees and to maximize the genetic gain in forest tree breeding program, providing a clue to determine the genetic marker-trait correlation. This review intorduces research on bark thickness and breeding strategy in Pinus elliottii, Pinus caribaea and their hybrid by Queensland Forest Service and ForBio Research Pty Ltd, University of Queensland, which employ RAPD technology. Genetic linkage map of $F_1$ hybrids includes 186 RAPD markers and 16 linkage groups (1641 cM long in total) and 6 quantitative trait loci are located putatively for bark thickness. Following recent research results and experiences in pine breeding programs, the forseeable stages in the application and development are proposed for marker assisted selectin; stage 1-determination of species specific markers for genes controlling traits of commercial interest, and stage 2-determination of marker-allele association for specific allelic variants within pure species. As pines inherit their megagametophytes from the seed parent and zygotic embryos from both male and female parents, the determination of marker-trait correlation is possible even in embryo stage, eventually making ways for the early selection of elite individuals.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.6
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• pp.655-660
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• 1998

To produce blastocysts more efficiently, it is required to identity accurately the factors involving embryonic cleavage in the chemically defined medium. Effects of pyruvate, lactate, calcium and protein sources on early cleavage of bovine follicular oocytes were investigated. The percentage of IVF embryos cleaved to ${\geq}$ 2-cell or ${\geq}$ 8-cell was higher in pyruvate (+) and lactate (+) (48 or 14%) than in pyruvate (-) and lactate (-) (22% or 4%), than in pyruvate (+) and lactate (-) (28% or 5%) and than in pyruvate (-) and lactate (+) (40% or 10%). Lactate was more effective than pyruvate during early cleavage of bovine embryos in the chemically defined medium. The percentage of IVF embryo cleaved to ${\geq}$ 2-cell and ${\geq}$ 8-cell in calcium (-) (19 and 6%) was significantly (p < 0.05) lower than in calcium (+) (78 and 45%). The percentage of embryos developed to ${\geq}$ 2-cell showed no significant (p < 0.05) difference among BSA, 1 and 20% FBS (57, 57 and 57%). Also the percentage of A grade embryos developed to ${\geq}$ 2-cell showed no significant (p < 0.05) difference among BSA, 1 and 20% FBS (40, 35 and 28%). The percentage of embryos developed to ${\geq}$ 8-cell showed no significant (p < 0.05) difference among BSA, 1 and 20% FBS (33, 23, and 22%). However, the percentage of A grade embryos developed to ${\geq}$ 8-cell in BSA (24%) was significantly (p < 0.05) higher than in 1 and 20% FBS (13 and 8%). The percentage of embryos developed to ${\geq}$ morula showed no significant (p < 0.05) difference among BSA, 1, 10 and 20% FBS (76, 76, 80 and 68%). The percentage of A grade embryos developed to ${\geq}$ morula in 10% FBS (59%) was significantly (p < 0.05) higher than 20% FBS (43%). The percentage of embryos developed to blastocyst showed no significant (p < 0.05) difference among BSA, 1, 10 and 20% FBS (34, 41, 43 and 32%). However, the percentage of A grade embryos developed to ${\geq}$ blastocysts in 10% FBS (25%) was significantly (p < 0.05) higher than in 20% FBS (8%).

• Korean Journal of Medicinal Crop Science
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• v.22 no.6
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• pp.423-428
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• 2014

This study was performed to identify optimal harvesting time of ginseng seeds and to examine the effect of $GA_3$ treatment for improvement of seed stratification rate. Ginseng seeds harvested from Land race, Chunpoong and Yunpoong cultivar in July 20 were tested for stratification rate. It was shown that stratification rates of land race, Yunpoong and Chunpoong cultivar were 94.1%, 93.1%, and 82.6%, respectively. Seeds of Chunpoong cultivar harvested 10-15 days later showed a comparable stratification rate to that of Land race, indicating that late harvest of Chunpoong seeds is beneficial for the increase of stratification rate. The higher stratification rate was found in mature seeds (92.3%) than immature seeds (37.8%), both of which were harvested in July 20. Stratification rate of mature seeds harvested in July 15 was 87.5%, demonstrating optimal harvesting time of ginseng seeds with higher stratification rate is after mid-July. An exponential growth of endosperms of ginseng seeds was observed from early June to mid-June and then slow growth was observed. There was no obvious growth of embryos from fertilization to mid-August. After the this time, embryos quickly grew until late October. Thus, appropriate stratification control is essential during the period (from early September to late October) in order to optimize embryo growth and development. While no increase of stratification rate was observed in seeds treated with 50 ppm of $GA_3$, significant increases were observed in seeds treated with 100 ppm of $GA_3$. At this concentration of $GA_3$, the stratification rate of Land race, Chunpoong and Yunpoong cultivar was 95.0%, 95.3%, and 96.5%, respectively.

• Development and Reproduction
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• v.13 no.4
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• pp.305-312
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• 2009

Multipotent spermatogonial stem cells (mSSCs), derived from uni-potent SSC, are a type of reprogrammed cells with similar characteristics to embryonic stem cells (ESCs). The aim of this study was to evaluate the potential for transgenesis of mSSC derived from outbred mice and the production of transgenic animal by the mSSC-insertion into embryo. mSSCs, established from outbred mice (ICR strain) in the previous study, were maintained and then transfected with a lenti-viral vector expressing green fluorescent protein (GFP), CS-CDF-CG-PRE. Embryonic stem cells (ESCs) were derived from inbred transgenic mice (C57BL/6-Tg (CAG-EGFP)) and were used as an experimental control. Transfected mSSCs were well proliferated in vitro and maintained their characteristics and normal karyotype. Ten to twelve mSSCs and ESCs were collected and inserted into perivitelline space of 8-cell mouse embryos, and then transferred them into uteri of poster mothers after an additional 2-days of culture. Percentage of mSSC-derived offsprings was 4.8% (47/980) and which was lower than those (11.7% (67/572)) of ESC-derived ones (P<0.05). However, even though different genetic background of mSSC and ESC origin, the production efficiency of coat-colored chimeric offspring in mSSC group was not different when compared it with ESC (6.4% (3/47) vs. 7.5% (5/67)). From these results, we confirmed that mSSC derived from outbred mice has a pluripotency and a potential to produce chimeric embryos or mice when reaggregatation with mSSC is performed.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2001.11a
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• pp.74-76
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• 2001

Comparing to mammals, male bird has the homozygote ZZ and female has the heterozygote n. Therefore, the sex of fertilized eggs is defined by female chromosome constitution. Although this cytological observation had been established, the molecular and cellular mechanism of germ cell differentiation are essentially unknown in aves. Especially, the differentiation of germ cells in mixed-sex chimeras has not yet been clearly elucidated. Primordial germ cells, which are the progenitors of sperm or egg after sexual maturity, firstly arise in the epiblast and migrate to embryonic gonads through the blood vessel. During the embryo development, these PGCs differentiate in the pathway of mate or female, respectively and develop the sperm or egg cells after sexual maturity. In this paper, we confirmed that the female PGCs could migrate into the recipient male gonads after transferring and differentiate into germ cells in the embryonic stages. The primordial germ cells were isolated from the female embryonic gonads of 5.5-day-old incubation and re-injected into the male recipient embryos of 2-day-old incubation, which produced mixed-sex chimera in the germline. The finding in this study demonstrated the ability of migration and differentiation of gonadal primordial germ cells in mixed-sex chicken.