• Development and Reproduction
• /
• v.24 no.4
• /
• pp.249-261
• /
• 2020

Spermatogonial stem cells (SSCs) have stemness characteristics, including germ cell-specific imprints that allow them to form gametes. Spermatogenesis involves changes in gene expression such as a transition from expression of somatic to germ cell-specific genes, global repression of gene expression, meiotic sex chromosome inactivation, highly condensed packing of the nucleus with protamines, and morphogenesis. These step-by-step processes finally generate spermatozoa that are fertilization competent. Dynamic epigenetic modifications also confer totipotency to germ cells after fertilization. Primordial germ cells (PGCs) in embryos do not enter meiosis, remain in the proliferative stage, and are referred to as gonocytes, before entering quiescence. Gonocytes develop into SSCs at about 6 days after birth in rodents. Although chromatin structural modification by Polycomb is essential for gene silencing in mammals, and epigenetic changes are critical in spermatogenesis, a comprehensive understanding of transcriptional regulation is lacking. Recently, we evaluated the expression profiles of Yin Yang 1 (YY1) and CP2c in the gonads of E14.5 and 12-week-old mice. YY1 localizes at the nucleus and/or cytoplasm at specific stages of spermatogenesis, possibly by interaction with CP2c and YY1-interacting transcription factor. In the present article, we discuss the possible roles of YY1 and CP2c in spermatogenesis and stemness based on our results and a review of the relevant literature.

• Korean Journal of Medicinal Crop Science
• /
• v.8 no.1
• /
• pp.41-48
• /
• 2000

This study was aimed to proliferate Gentiana axillariflora Leveille which is one of the important medicinal and ornamental plants, by establishment of multiple shoot formation and embryogenesis through tissue culture technique. Callus was formed on MS (Murashige and Skoog) medium supplemented with 2,4-D, CPA, but not formed with BAP. The addition of 2,4-D 2 mg/ l into the medium was effective for callus formation and the rate of callus formation was about 90%. Somatic embryos were obtained on MS medium for two months. When callus was cultured on MS medium with combination treatment of 2,4-D 0.5 mg/ l and BAP 0.5 mg/ l, the number of embryo formed was better than that of other single or combination treatments and the total numbers of embryo a were 18.8 (number of total embryo/number of explants incubated = 753/40) at mean. Callus induction from stem and node explants was increased by addition of TDZ 2 mg/ l in the presence of 2,4-D 2 mg/ l, respectively. The best result about the differentiation of shoots was obtained on MS medium added BAP 2 mg/ l from node culture. Multiple shoots from shoot apex were induced on MS medium containing BAP 1 mg/ I and TDZ 1 mg/ l , BAP 2 mg/ l and TDZ 1 mg/ l. The number of multiple shoots per one explant was above seventy plants. It was the most effective regeneration system for rapid multiplication of Gentiana axillariflora Leveille.

• Journal of Embryo Transfer
• /
• v.26 no.1
• /
• pp.79-84
• /
• 2011

This study was performed to identify the differentially methylated region (DMR) and to examine the mRNA expression of the imprinted H19 gene in day 35 of SCNT pig fetuses. The fetus and placenta at day 35 of gestation fetuses after natural mating (Control) or of cloned pig by somatic cell nuclear transfer (SCNT) were isolated from a uterus. To investigate the mRNA expression and methylation patterns of H19 gene, tissues from fetal liver and placenta including endometrial and extraembryonic tissues were collected. The mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. Bisulfite analyses demonstrated that the differentially methylated region (DMR) was located between -1694 bp to -1338 bp upstream from translation start site of the H19 gene. H19 DMR (-1694 bp to -1338 bp) exhibits a normal mono allelic methylation pattern, and heavily methylated in sperm, but not in oocyte. In contrast to these finding, the analysis of the endometrium and/or extraembryonic tissues from SCNT embryos revealed a complex methylation pattern. The DNA methylation status of DMR Region In porcine H19 gene upstream was hypo methylated in SCNT tissues but hypermethylated in control tissues. Furthermore, the mRNA expression of H19 gene in liver, endometrium, and extraembryonic tissues was significantly higher in SCNT than those of control (p<0.05). These results suggest that the aberrant mRNA expression and the abnormal methylation pattern of imprinted H19 gene might be closely related to the inadequate fetal development of a cloned fetus, contributing to the low efficiency of genomic reprogramming.

• Korean Journal of Plant Tissue Culture
• /
• v.25 no.1
• /
• pp.13-19
• /
• 1998

We have reported previously that intact embryogenic cells can be used instead of protoplasts for electroporation-mediated transformation of zoysiagrass and rice. In this study, conditions of the tissue electroporation were examined to optimize the procedures. Embryogenic cell suspensions were established in liquid MS medium containing 2 mg/L of 2,4-D with embryogenic calluses induced from mature embryos of Z. japonica. The suspension-cultured cell clumps were electroporated with 35S-gusA expression vector DNA, and degrees of DNA introduction into the cells were determined by histological expression rates of the gusA marker gene. DNA transfer into the cell clumps occurred in wide range of voltage (100-400 V) and capacitance (10-1980 $\mu\textrm{F}$), but more in the ranges of 200-300 V and 330-800 $\mu\textrm{F}$ DNA concentrations higher than 6 $\mu\textrm{g}$/mL were adequate for GUS expression of the electroporated cells. DNA transfers were confirmed in all three embryogenic cell lines but only in one out of eleven non-embryogenic lines. Positive GUS expressions occurred with DNAs added even 20-40 h after pulse treatments. As a promoter of gusA, Act1 and Ubi1 were effective 7 and 5 times than 35S respectively in number of GUS expression units on electroporated cell clumps. Embryogenic cell clumps survived and regenerated into plantlets after pulse treatments of wide range of conditions.

• Korean Journal of Medicinal Crop Science
• /
• v.3 no.3
• /
• pp.207-216
• /
• 1995

This study was conducted to establish technologies for plug seedling production using somatic embryos-derived regenerants and their seeds in Bupleurum falcatum L. Among distilled water, GA (0.1mg/l) and putrescine (0.lmg/l) treated to regenerants for acclimatization, GA was most effective to develop shoots and roots, 1/2X MS medium and NAA 0.1mg/l + BA 0. 5mg/l enhanced the growth rates of the regenerants and increased dry weight. Activated charcoal effected to grow markedly leaves and roots of the regenerants at the level of 0.4 %. Regenerants increased their plant height, root length and dry weight at $30^{\circ}C$. Plug seedlings originated from seeds of the tissue culture regenerants showed the maxium growth on the mixture of peatmoss soil (2) and mountain sand (1) .Root length, leaf area and dry weight of plug seedlings increased significantly when No.1, 2 and 3 of Wondergrow solution were mixed in the ratio of 1.3 - 0.9 - 0.1. Light supplement (4%) and high tem­perature $(30^{\circ}C)$ promoted the growth of plug seedlings as well as dry weight. Ninety days seedlings were more vigorous and adaptable for transplanting than other seedlings.

• Journal of Plant Biotechnology
• /
• v.36 no.1
• /
• pp.75-80
• /
• 2009

In order to develop transgenic sweetpotato plants [Ipomoea batatas (L.) Lam. cv. Yulmi] with enhanced tolerance to oxidative stress, we constructed transformation vectors expressing 2-Cys peroxiredoxin (Prx) gene under the control of the stress-inducible SWPA2 or enhanced 35S promoter (named as SP or EP). Transgenic sweetpotato plants were attempted to generate from embryogenic calli using an Agrobacterium-mediated transformation system. Embryogenic calli gave rise to somatic embryos and then converted into plantlets on MS medium containing 100 mg/L kanamycin. Transgenic plants were regenerated in the same medium. Southern blot analysis confirmed that the Prx gene was inserted into the genome of the plants. To further study we selected the transgenic plant lines with enhanced tolerance against methyl viologen (MV). When sweetpotato leaf discs were subjected to methyl MV at $20{\mu}M$, transgenic plants showed about 40% higher tolerance than non-transgenic or empty vector-transformed plants.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2010.05a
• /
• pp.12-12
• /
• 2010

We developed droplet-vitrification protocol, a combination of droplet-freezing and solution-based vitrification, and applied to germplasm collections of garlic, potato, lily as well as cell lines, including hairy roots, somatic embryos. To establish a garlic cryobank, four Korean garlic field collections at Danyang, Suwon, Mokpo and Namhae were cryopreserved last five years. The protocol applied consisted of preculture for 3-4 days at $10^{\circ}C$ on solid MS medium with 0.3M sucrose, loading for 40 min in liquid medium with 35% PVS3, dehydration with PVS3 for 150 min, cooling in $5{\mu}l$ droplets of PVS3 placed on aluminum foil strips by dipping these strips in liquid nitrogen, warming them by plunging the foil strips into pre-heated($40^{\circ}C$) 0.8M sucrose solution for 30s. A total of over 900 accessions of garlic were stored in liquid nitrogen for long-term conservation using unripe inflorescences, cloves or bulbils. Twelve alternative plant vitrification solutions were designed by modifying cryoprotectant concentrations from the original PVS2 and PVS3. The results suggest that PVS2-based vitrification solutions with increased glycerol and sucrose and/or decreased DMSO and EG concentrations can be applied for medium size explants which are tolerant to chemical toxicity and moderately sensitive to osmotic stress. PVS3 and variants can be used widely when samples are heterogeneous, of large size and/or very sensitive to chemical toxicity and tolerant to osmotic stress.

• Plant Resources
• /
• v.5 no.1
• /
• pp.29-44
• /
• 2002

In this study, plant regeneration through in vitro culture from plantlet stems of Yooja (C. junos Sieb.) and trifoliate orange (P. trifoliata Rafin.) was attempted to make mass-production system of virus-free plants having the same genotype with mother plant. In order to investigate physiological change depending on the developmental stage of plant regeneration, the changes of total protein, peroxidase and esterase activity and their isozyme patterns as well were examined in 1/2 MS medium. The results are as follows : 1. The MS medium for the optimal callus induction and shoot formation was utilized. The medium was supplemented either with 2,4-D and Kinetin or with BA and NAA. The optimal concentrations were the combination of 1.0mg/ 2,4-D +0.3mg/ Kinetin and 1.0mg BA +0.3mg NAA in callus induction and shoot formation, respectively. 2. For the plant regeneration from somatic embryos, 1/2 MS medium was used with supplements of growth regulators (free, 1.0mg/ IBA +1.0mg/ BA ,0.5mg/ IBA +0.5mg/ BA). Shooting and rooting were the best in the treatment of 0.5mg/ IBA and 0.5mg/ BA combination. 3. The total protein content has a tendency of increase with the developmental stage of embryo, but it was decreased at the plantlet. Also it was the highest at 8 and 6 weeks stage in C. junos Sieb. and P. trioliata Rafin, respectively. In the SDS-PAGE pattern of protein, C. junos Sieb. showed bands of 29.0 and 40kDa at 10 weeks. The 45,66 and 97.4 kDa bands at 10 weeks of culture were shown in P. trifoliata Rafin. 4. The highest esterase activity was shown at the 6 and 8 weeks of culture in C.junos Sieb. and P. trifoliata Rafin.., respectively. 5. Esterase isozyme patterns were shown difference according to the developmental stage. In C. junos Sieb. a new band was observed at pl 7.7 following 4 weeks culture. On the other hand, new bands in P. trifoliata Rafin. were observed at pl 7.5~6.5 following 4 and 6 weeks culture, respectively.

• Korean Journal of Environment and Ecology
• /
• v.35 no.3
• /
• pp.285-293
• /
• 2021

This study was conducted to elucidate spawning period characteristics and early life development of eight barbel loach, Lefua costata (Balitoridae) at the Jusucheon stream, Okgye-myeon, Gangneung-si, Gangwon-do, Korea between January and December in 2018. The spawning period was estimated to be from May to August, considering the change in the gonad-somatic index, the appearance of young fry, and frequency distribution in egg diameter. It was a multi-spawning type. The gender ratio was 1:0.79 with 1,117 females and 879 males collected. The egg size was 0.24-0.93 mm, with mature and immature eggs found during the spawning period. The size of mature eggs was 0.71±0.02 mm, and the average number of fecundity was 1,786±818 (n=31). Observation of the egg development showed that the fertilized eggs were the sticky, gray, segregated, and demersal type with 0.76±0.03 mm in diameter. The hatching of the embryos began at about 34 hours (hatching rate 50%) after fertilization underwater temperature of 25℃. The average length of the newly hatched pre-larvae was 2.7±0.11 mm. The average length of pre-larvae at 4 days after hatching was 4.5±0.16 mm, and the yolk sac was completely absorbed and entered the post-larvae stage. At 20 days after hatching, the average length of post-larvae was 11.5±0.67 mm, and their fin rays were formed before they transitioned to the juvenile stage. At 100 days after hatching, the average length reached 49.8±2.60 mm, and the appearance and the lateral sideband patterns were similar to those of the adult fish.

• The Korean Journal of Malacology
• /
• v.24 no.2
• /
• pp.153-159
• /
• 2008

We investigated artificial mass seed production of a Chinese scallop, Patinopecten yessoensis, in 2004. The GSI(gonad somatic index) of the Chinese scallop, P yessoensis was 17.2 on mid-February, 20.2 on mid-March, while that of Korean scallop, P yessoensis was 6.9 on mid-February, 10.8 on mid-March. Matured 120 females and 350 males were selected for artificial mass production. They were exposed in air for 1 hr at over $20^{\circ}C$, and placed into a spawning tank(20 ton) containing sea water treated with UV radiation at $12^{\circ}C$. We gained a total of 228,000 thousand scallop embryos between March 10th and 15th, and reared larvae at the indoor tank during 25 days. When the mean shell length of larvae reached 250 ${\mu}m$ and they have eye-spots, the number of pre-settling larvae was 47,500 thousand. We gained 1,850 thousand attached scallop spats from two kinds of collectors. Attached spats were reared in indoor tank for different periods from 5 days to 60 days. They were divided into 5 groups according to the length of reared days. Each group of attached spats was moved to intermediate rearing sites at Yangyang fishing port in Gangreung-city for acclimation to ocean environments. The highest survival rate of attached spats was 13.0% shown at the group reared for 12 days, but the significant difference in their growth was not found between the groups. The shell length of artificial attached spats increased from 0.9 ${\mu}m$ on July 10th to 24.7 ${\mu}m$ on December 16th with the survival rate of 85.0% while that of natural attached spats increased from 0.6 ${\mu}m$ on July 10th to 23.9 ${\mu}m$ on December 16th with the survival rate of 85.7%.