• FLOWER RESEARCH JOURNAL
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• v.19 no.4
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• pp.225-230
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• 2011

This research was performed for making data-base of cross-breeding between Cymbidium goeringii cultivars. Morphological characteristics were investigated and then genetic relationship was analyzed. Collected 20 Cymbidium goeringii cultivars were clustered into 2 groups. Seven cultivars were clustered into group I, and thirteen cultivars were clustered into group II. Group I doesn't have leaf pattern. Group have leaf pattern. The genetic relationship among collected 20 Cymbidium goeringii cultivars was anaylzed using RAPD with ten 10-mers random primer. Eighty-nine bands were generated by RAPD. Among the rest, three bands were monomorphic and eight-six bands were polymorphic. Overall similarity degree ranged from 0.521 to 0.862. The result of RAPD analysis was clustered into 2 groups, too. Sixteen cultivars were clustered into GroupX, and four cultivars were clustered into GroupY. Result of classification with morphological characteristics and RAPD showed different pattern, but 4 cultivars of GroupY by RAPD analysis were included in groupby morphological characteristics. Crossbreeding combination among low related coltivars in RAPD analysis may get more efficient result.

• Korean Journal of Breeding Science
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• v.41 no.1
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• pp.1-8
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• 2009

This study was carried out to breed and develop high quality and functional nutrient tomato with multi disease resistance as well as a stable growing adaptation for fresh market usage under protected plastic houses cultivation. The materials were used 5 inbred lines and their 6 hybrids of large tomato group, which have been bred and developed from 1999 to 2007 in Division of Plant Resource Department of Chungnam National University. Fruit weight showed hybrid vigor effect that $F_1$ hybrids weighed more than their parent lines, fruit shape formed three type of oblate, deep oblate and globe shape, in firmness and pericarp thickness have got a high significant correlation, inbred DN611 line was measured the most firm fruit with 6.04 mm pericarp thickness. In fruit color at maturity, pink color crossed to red color appeared all red fruit color in the $F_1$ hybrids, it means red skin color is a dominant gene compared to pink skin color is a recessive gene in tomato, while between fruit skin color and shoulder part color showed no any co-relationship. The sugar content and titratable acid of $F_1$ hybrids inherited an intermediate data of their parent lines, the flavor of KP543 inbred line and the hybrid (JB535 x KP543) revealed the better taste with high brix and proper titratable acid content$^{*}$. In beta carotene content DN611 line showed 2~3 times higher than other materials so that its 3 hybrids contained an increased level of beta carotene, lycopene content was not so much difference among inbred lines and $F_1$ hybrids, of them MD508 contained higher of 8.72 mg and hybrid (JB535 x JA517) had 8.05 mg lycopene content per 100 g fruit, overall pink skin color and red skin color measured a higher lycopene content than yellow and orange skin color at ripe stage. In disease resistance test by PCR marker for Fusarium race2 (I2), Nematode (Mi1), ToMV ($Tm2^2$), Cladosporium (Cf9), (JB535 x JA517) hybrid have got multi-resistance with homozygote band in Nematode, ToMV, Cladosporium and heterozygote band in Fusarium race2. Through this breeding program we could select high quality and functional nutrient with multi resistant $F_1$ hybrids and inbred lines in tomato which are two best hybrids (JB535 x MD508), (JB535 x JA517), additionally developed high beta carotene inbred line DN611 and increased the level of lycopene inbred line MD508. These results will be very useful to make a high quality tomato variety continuously.

• Development and Reproduction
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• v.22 no.4
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• pp.379-391
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• 2018

Through the development of organic synthetic skill, chemicals that mimic signaling mediators such as steroid hormones have been exposed to the environment. Recently, it has become apparent that this circumstance should be further studied in the field of physiology. Estrogenic action of chronic low-dose nonylphenol (NP) and di-(2-ethylhexyl) phthalate (DEHP) in mouse uterus was assessed in this study. Ten to twelve-week-old female mice (CD-1) were fed drinking water containing NP (50 or $500{\mu}g/L$) or DEHP (133 or $1,330{\mu}g/L$) for 10 weeks. Uterine diameter, the thickness of myometrium and endometrium, and the height of luminal epithelial cells were measured and the number of glands were counted. The expression levels of the known $17{\beta}$-estradiol ($E_2$)-regulated genes were evaluated with real-time RT-PCR methodology. The ration of uterine weight to body weight increased in $133{\mu}g/L$ DEHP. Endometrial and myometrial thickness increased in 133 and $1,330{\mu}g/L$ DEHP treated groups, and in 50, $500{\mu}g/L$ NP and $133{\mu}g/L$ DEHP, respectively. The height of luminal epithelial cell decreased in NP groups. The numbers of luminal epithelial gland were decreased in NP groups but increased in $50{\mu}g/L$ DEHP group. The histological characters of glands were not different between groups. The mRNA expression profiles of the known $17{\beta}$-estradiol ($E_2$) downstream genes, Esr1, Esr2, Pgr, Lox, and Muc1, were also different between NP and DEHP groups. The expression levels dramatically increased in some genes by the NP or DEHP. Based on these results, it is suggested that the chronic low-dose NP or DEHP works as estrogen-like messengers in uterus with their own specific gene expression-regulation patterns.

• Journal of Food Hygiene and Safety
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• v.34 no.5
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• pp.502-507
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• 2019

Cephalopods are one of the most important fishery resources in the world because of their desirable taste and nutritional value. In south Korea, one of the countries in which a large amount of seafood is consumed, cephalopods (e.g., octopus, squid, and cuttlefish) have an annual consumption rate of over 400,000 metric tons. In this study, octopus and squid products (n=28) sold on the market were monitored by analyzing sequences of DNA barcode markers (cytochrome c oxidase subunit I and 16S ribosomal RNA genes). For species identification, the NCBI BLAST database was screened with the sequences and analyzed as a query. In this BLAST search, twelve squid products showed 99-100% sequence identity to Dosidicus gigas (n=3) and Todarodes pacificus (n=9). In the case of the other 16 products that were declared using octopus as raw materials on the labels, six products were identified as Cistopus taiwanicus (n=1), Amphioctopus marginatus (n=1), Scaeurgus unicirrhus (n=1), and Dosidicus gigas (n=3). Monitoring results indicated that a significant percentage (37.5%) of mislabeling was present in octopus products sold on the South Korean market.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.2
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• pp.133-147
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• 2005

Objectives: This study was carried out to establish human embryonic stem cells derived from frozen-thawed embryos using mouse embryonic fibroblasts (mEFs), human fetal skin and muscle fibroblasts as feeder cells, and to identify the characteristic of embryonic stem cells. Methods: When primary mEFs, human fetal skin and muscle fibroblasts were prepared, passaging on 4 days from replating could have effective trypsinization and clear feeder layers. Eight of 23 frozenthawed 4~8 cell stage embryos donated from consenting couples developed to blastocysts. Inner cell mass (ICM) was isolated by immunosurgery. ICM was co-cultured on mEFs, human fetal skin or muscle fibroblasts. The ICM colonies grown on mEFs, human fetal skin or muscle fibroblasts were tested the expression of stage specific embryonic antigen-3, -4 (SSEA-3, -4), octamer binding transcription factor-4 mRNA (Oct-4) and alkaline phosphatase surface marker. Results: We obtained 1 ICM colony from 2 ICM co-cultured on mEFs as feeder cells and did not obtain any ICM colony from 6 ICM clumps co-cultured on human fetal skin or muscle fibroblasts. The colony formed on mEFs could be passaged 30 times every 5 days with sustaining undifferentiated colony appearance. When the colonies cultured on mEFs were grown on human fetal skin or muscle fibroblasts, the colonies could be passaged 15 times every 9 days with sustaining undifferentiated colony appearance. The colonies grown on mEFs and human fetal fibroblasts expressed SSEA-4 and alkaline phosphatase surface markers and positive for the expression of Oct-4 by reverse transcription-polymerase chain reaction (RT-PCR). The produced embryoid body differentiated spontaneously to neural progenitorlike cells, neuron-like cells and beating cardiomyocyte-like cells, and frozen-thawed embryonic stem cells displayed normal 46,XX karyotype. Conclusions: The human embryonic stem cells can be established by using mEFs and human fetal fibroblasts produced in laboratory as feeder cells.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.2
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• pp.121-132
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• 2005

연구목적: 본 연구에서는 비폐쇄성 무정자증 환자에서 나타나는 정자형성과정의 이상과 고환세포의 세포자연사와의 연관관계 여부를 확인하였다. 또한 SELDI-TOF MS 분석을 통하여 고환 내 단백질 발현 양상을 확인하고, 질환에 따른 효과적인 biomarker 개발 가능성 여부를 확인하였다. 재료 및 방법: RT-PCR 및 면역조직화학법을 사용하여 고환에서의 Fas, FasL, Bcl-2, Bax와 Caspase-3의 발현 양상을 확인하고, in situ DNA 3'-end-labelling 방법으로 고환세포의 세포자연사 양상을 확인하였다. SELDI-TOF MS 분석법에 의한 고환의 병리학적 소견에 따른 단백질 발현 변화는 소수성 칩 ($H_4$)을 사용하여 분자량 10~100 kDa 범위 내에서 분석하였다. 결 과: 정상적인 정자형성과정을 보이는 폐쇄성 무정자증 환자의 고환에 비해 지주세포 증후군 (Sertoli cell only syndrome)과 성숙정지 (maturation arrest)를 보이는 고환 내 생식세포와 지주세포에서 세포자연사가 현저하게 증가한 것을 확인할 수 있었다. 세포자연사 관련인자들의 발현 양상을 확인한 결과, 지주세포 증후군과 성숙정지 환자군에서 Fas와 FasL mRNA의 발현이 증가하였으나, bcl-2, bax와 caspase-3 mRNA 발현의 경우에는 두 질환 모두에서 유의한 차이를 확인할 수 없었다. FasL 단백질 발현의 경우, 세포자연사의 증가가 관찰되었던 지주세포 증후군과 성숙정지를 보이는 환자의 간질세포와 지주세포에서 증가하는 양상을 나타내었다. SELDI-TOF MS 분석 결과에서 폐쇄성 무정자증 환자군에 비해 전체적인 단백질 발현양이 지주세포 증후군과 성숙정지 환자의 고환에서 감소하는 양상을 보였으며, 특히, 16.730 kDa 단백질의 현저한 감소를 확인할 수 있었다. 결 론: 본 연구결과를 통해 비폐쇄성 무정자증 환자에서 나타나는 정자형성과정의 장애는 생식세포의 비정상적인 세포자연사와 연관되어 있으며, 고환 내 Fas와 FasL의 비정상적인 발현이 주된 원인인 것을 확인할 수 있었다. 또한, SELDI-TOF MS 분석법을 통한 단백질 발현 양상의 연구는 무정자증 환자에서의 다양한 병리학적 소견을 쉽게 파악할 수 있는 biomarker 발굴뿐만 아니라 질환의 원인규명을 위한 연구에도 유용하게 이용될 수 있을 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.3
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• pp.279-286
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• 2005

Objective: Although several genetic factors have been associated with defects in human spermatogenesis, the unambiguous causative genes have not been elucidated. The male infertility by haploinsufficiency of PRM1 or PRM2 has been reported in mouse model. The aim of this study was to identify the single nucleotide polymorphisms (SNPs) of PRM1 and PRM2, related to the genotype of Korean infertile men. Methods: Genomic DNAs were extracted from peripheral bloods of infertile men with oligozoospermia or azoospermia, and analyzed using polymerase chain reaction (PCR) and direct sequencing. We carried out the direct sequencing analysis of amplified fragments in PRM1 (557 nucleotides from -42 to 515) and PRM2 (599 nucleotides from 49 to 648) genes, respectively. Results: Three SNPs of coding region in the PRM1 gene was found in the analysis of 130 infertile men. However, the SNPs at a133g (aa 96.9%, ag 3.1% and gg 0.0%), c160a (cc 99.2%, ca 0.8% and aa 0.0%) and c321a (cc 56.9%, ca 35.4% and aa 7.7%) coded the same amino acids, in terms of silence phenotypes. On the other hand, as results of the PRM2 gene sequencing in 164 infertile men, only two SNPs, g398c (gg 62.2%, gc 31.1% and ga 6.7%) and a473c (aa 63.4%, ac 29.9% and cc 6.7%), were identified in the intron of the PRM2 gene. Conclusions: There was no mutation and significant SNPs on PRM1 and PRM2 gene in Korean infertile men. These results suggest that the PRM1 and PRM2 genes are highly conserved and essential for normal fertility of men.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.3
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• pp.207-216
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• 2005

Objective: To understand the crucial requirement for the normal early folliculogenesis, we evaluated molecular as well as physiological differences during in vitro ovarian culture. Among the important regulators for follicle development, anti-Müllerian hormone (AMH) and FSH Receptor (FSHR) have been known to be expressed in the cuboidal granulosa cells. Meanwhile, it is known that c-kit is germ cell-specific and GDF-9 is also oocyte-specific regulator. To evaluate the functional requirement for the competence of normal follicular development, we investigated the differential mRNA expression of several factors secreted from granulosa cells and oocytes between in vivo and in vitro developed ovaries. Materials and Methods: Ovaries from ICR neonates (the day of birth) were cultured for 4 days (for primordial to primary transition) or 8 days (for secondary follicle formation) in ${\alpha}$-MEM glutamax supplemented with 3 mg/ml BSA without serum or growth factors. The mRNA levels of the several factors were investigated by quantitative real-time PCR analysis. Freshly isolated 0-, 4-, and 8-day-old ovaries were used as control. Results: The mRNA of AMH and FSHR as granulosa cell factors was highly increased according to the ovarian development in both of 4- and 8-day-old control. However, the mRNA expression was not induced in both of 4- and 8-day in vitro cultured ovaries. The mRNA expression of GDF-9 known to regulate follicle growth as an oocyte factor was different between in vivo and in vitro developed ovaries. In addition, the transcript of GDF-9 was expressed in the primordial follicles of mouse ovaries. The mRNA expression of c-kit was not significantly different during the early folliculogenesis in vitro. Conclusion: This is the first report regarding endogenous AMH and FSHR expression during the early folliculogenesis in vitro. In conclusion, it will be very valuable to evaluate cuboidal granulosa cell factors as functional marker(s) for normal early folliculogenesis in vitro.

• The Korean Journal of Malacology
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• v.29 no.3
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• pp.171-179
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• 2013

Arginine kinase (ArK) is known to play an important role in most invertebrates the level of ATP by phosphorylation of phosphagens in cell and immuninty in living organisms. ArK has been identified in many kinds of organisms ranging from invertebrate to vertebrate. However, no ArK gene has been cloned and investigated from N. samarangae. This leads us to identify ArK cDNA (NsArK) from the expressed sequence tag (EST) sequencing of N. samarangae. Sequence analysis indicated that the coding region of 1,065 bp contains 355 amino acid residues. Molecular phylogenetic analysis shows that NsArK had very high similarities with mollusca and arthropoda. In an attempt to investigate a potential role of NsArK in the digestive gland of N. samarangae, expression patterns were analyzed. RT-PCR analsysis shows that NsArK mRNA is induced in the rane of 1.2 fold at 6 hr by laminarin when compared with the control. The immunnologial and physiological role of NsArK remains to be further investigated in N. samarangae.

• Asian Journal of Turfgrass Science
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• v.23 no.2
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• pp.307-316
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• 2009

Creeping bentgrass (Agrostis palustrics Huds.) is cool season turfgrasse that is used for putting green in golf course. Creeping bentgrass cultivars are difficult to distinguish with the same species because of similar morphological characters and low level of genetic diversity. The SCAR markers using the specific DNA can be useful for differentiating between creeping bentgrass cultivars. Five RAPD primers were used for specific band detection among creeping bentgrass cultivars, penncross, penn A-4, crenshaw, L-93, CY-2, T-1. The pairs of SCAR primers for six cultivers were designed by the specific sequences of the bands that amplified by RAPD. Three of the six SCAR primers could not make the use as SCAR primers because the specific false bands were detected in all cultivars. The remaining pairs of SCAR primer, CY850F/R, T700F/R, L2900F/R, amplified the specific band at expected size for three cultivars, CY-2, T-1, L-93, respectively. The CY850F/R primer amplified a band of 850bp in CY-2 cultivar, the T700F/R primer amplified a band of 700bp in T-1 cultivar, and the L2900F/R primer amplified a band of 2.9kb in L-93 cultivar. In this study we developed the SCAR markers to identify and distinguish the inerseeded creeping bentgrass cultivars in a golf course green.