Background : Platycodon grandiflorum is a perennial plant and a member of Camanulaceae family. Since ancient times, they have been using P. grandiflorum as an important medicinal plant in Korea. Platycodin D is the most abundant saponin derived from P. grandiflorum and pharmacologically active component. UDP-glycosyltransferases (UGTs) are important enzymes in the saponin biosynthesis. UGT is a glycosyltransferase and act on the final step of the secondary metabolite biosynthesis. Methods and Results : We tried to identify UGT genes related to saponin biosynthesis of P. grandiflorum through RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obtained 122,663 contigs and found 137 putative UGT genes. Familes of UGT71, UGT73, and UGT74 were selected as putative saponin biosynthesis related gene families using phylogenetic relationship analysis. qPCR condition about UGT73 is preheating 94℃ 180 sec, denaturation 94℃ 60 sec, annealing 53℃ 60 sec, extension 72℃ 90 sec, final extension 72℃ 600 sec, 45 cycles repeated. Conclusion : The results in this study could help to find the UGTs related to saponin biosynthesis pathway of P. grandiflorum.

Background : Astragalus membranaceus is one of the most widely used traditional medicinal herbs in Korea. Studies on the genomic of A. membranaceus resources have not been carried out so far. The present study was carried out to discriminate A. membranaceus based on genetic diversity using genomic simple sequence repeat (SSR) markers. Methods and Results : We collected 5 A. membranaceus lines: Asung, Poongsung, Am-Jecheon, Am-Sancheong, and Am-China. One hundred mg of fresh leaves were used for genomic DNA extraction using the DNeasy plant DNA isolation kit (Qiagen GmbH, Hilden, Germany). 450,449 contigs were searched for 147,766 SSR candidate loci in this study using the MicroSAtellite identification tool (MISA). We selected 949 A. membranaceus genomic SSR markers that were showed variation for the five collections in silico screening with CLC genomics workbench program. The genetic diversity of all A. membranaceus resources was analyzed using 17 SSR markers employing the DNA fragment analysis method. Based on the genetic diversity analysis, these lines were classified into four distinct groups. Conclusion : These findings could be used for further research on cultivar development using molecular breeding techniques and for conservation of the genetic diversity of A. membranaceus. Furthermore, the markers could be used for marker-assisted selection for crop breeding.

Background : Codonopsis lanceolata is used as a natural medicine or vegetables. It originates in East Asia such as Korea, Japan and China. C. lanceolata roots contain various chemical compounds including saponins like Panax ginseng. Although C. lanceolata are cultivated in different regions of South Korea, no variety has been developed. Therefore, it is necessary to develop discriminating methods such as molecular markers in C. lanceolata species. Methods and Results : To find simple sequence repeat (SSR) markers, we sequenced C. lanceolata genomic DNA using Illumina HiSeq 2000 System. A total of 250,455 putative SSR loci were obtained, and 26,334 non-redundant primers were designed to amplify these SSRs. Di-nucleotied repeats were the most abundant SSR reapeats, accounting for 89.53% (23,578) of primer designed SSRs. Tri-nucleotide, tetra-nucleotide and penta-nucleotide accounted for 8.44% (2,223), 1.3%, (348) 0.2% (55), respectively. Tri-, tetra-, and penta-nucleotide (total of 2,626 SSRs) were investigated in silico to identify polymorphism between individuals. Consequently, 573 SSRs showed polymorphism. Forty genomic SSR markers were tested in 16 C. lanceolata plants for determination of PCR amplification and polymorphism. From these primers, 27 (67.5%) amplified products and the average polymorphism information content (PIC) value was 0.52. Conclusion : We development 27 SSR markers from C. lanceolata using NGS, and it could be used for breeding of new varieties in the future.

Background : Angelica gigas is a monocarpic biennial or short lived perennial plant. A. gigas, also called Dang Gui or Korean Angelica, is a major medicinal herb used in Asian countries such as Korea, Japan and China. In Korea, we are using the roots of A. gigas, but, they are using Angelica sinensis in China and Angelica acutiloba in Japan to obtain many active constituents. The biggest problem in the using of A. gigas would be the confusion with A. acutiloba or A. sinensis. These three plants can't be distinguished by appearance. And the constituent ratios of the three plants are different. This confusion can cause an accident or the pharmaceutical effects do not meet the expectations. In this study, we developed chloroplast SSR markers that can distinguish A. gigas, A. acutiloba and A. sinensis. Methods and Results : We collected A. gigas, A. acutiloba and A. sinensis. and extrated DNA using CTAB method. The DNA was diluted to 10 ng/㎕ and kept -20℃. We designed the primer sets using CLC Main Workbench based on chloroplast DNA SSR region of A. gigas. PCR were performed on the three angelica plant samples (in 5 repeat). Conclusion : We made five primer sets from five SSR regions of A. gigas cpDNA. All of the primer sets amplified the amplicon effectively. Two of the 5 primer sets had polymorphism. We can distinguish A. gigas, A. acutiloba, and A. sinensis using the 2 primer sets

Twenty apple germplasm accessions from the Korean Genebank were successfully cryopreserved using two-step freezing to back up genetic resources maintained by field collections. This study examined the morphological and genetic stability of cryopreserved dormant apple buds that were stored in liquid nitrogen, and then rewarmed and regrown. Whole plants were regenerated directly from dormant buds through budding without an intermediary callus phase. The cryopreserved buds produced high levels of shoot formation (76.2-100%), similar to those of noncryopreserved buds (91.3-100%), with no observed differences between cryopreserved and noncryopreserved materials. Three of the twenty cryopreserved apple germplasm accessions were used to assess morphological and genetic stability. No differences in morphological characteristics including shoot length, leaf shape, leaf width/length ratio, and root length were observed between controls (fresh control and noncryopreserved) and cryopreserved plantlets. The genetic stability of regenerants (before and after cryopreservation) was investigated using inter simple sequence repeat (ISSR) markers. The ISSR markers produced 253 bands using four primers, ISSR 810, SSR 835, ISSR 864, and ISSR 899. These markers showed monomorphic banding patterns and revealed no polymorphism between the mother plant and regenerants before and after cryopreservation, suggesting that cryopreservation using two-step freezing does not affect the genetic stability of apple germplasm. These results show that two-step freezing cryopreservation is a practical method for long-term storage of apple germplasms.

We micropropagated pear (Pyrus species) using shoot tips and nodal explants from three pear genotypes. The ability to establish shoot tip cultures, proliferate shoots, induce rooting, and acclimatize the resulting plantlets are all elements of in vitro micropropagation. Shoots were induced from shoot tips on Murashige and Skoog medium (MS) with five different plant growth regulator combinations. The highest shoot formation rates were achieved for the three genotypes using MS supplemented with 1.0 ㎎/L N 6 -benzyladenine (BA) and 0.1 ㎎/L gibberellic acid (GA3). The maximum shoot number and shoot length for the three cultivars were recorded with 2.0 ㎎/L BA and 0.2 ㎎/L indole-3-butyric acid (IBA) in multiplication medium using nodal explants produced from microshoots. Nodal explants with one or two axillary buds cultured for three weeks initiated roots on medium supplemented with various concentrations of 1-naphthaleneacetic acid (NAA) or/and IBA in half-strength MS medium for adventitious rooting. The highest rooting response was with the combination of 0.2 ㎎/L NAA and 0.2 ㎎/L IBA. A combination of NAA and IBA resulted in a significant increase in the rooting ratio over NAA or IBA alone. In this medium, the root formation rate according to ranged from 68.9% for the BaeYun No. 3 genotype to 51.8% for the Hwanggeum genotype. We also investigated the influence of the concentration the polyamine phloroglucinol in rooting medium. For all three genotypes, the highest rooting ratio, longest root length, and greatest root number were observed in the treatments with 75-150 ㎎/L phloroglucinol. Most rooted plants were acclimatized successfully.

The pregnancy and abortion process involves a complex mechanism with various immune cells present in the implantation sites and several hormones associated with pregnancy, such as leptin, ghrelin and nesfatin-1. However, the mechanism underlying spontaneous abortion by maternal T helper 17 (Th17) present in the implantation sites and nesfatin-1, which is of anorexigenic hormones, is not fully understood so far. Therefore, the purpose of this study was to examine the possible roles of Th17 cells present in the implantation sites and nesfatin-1 expressed in the uterus on spontaneous abortion using the CBA/j × DBA/2 mouse model. Th17 transcription factor, ROR-γt mRNA expression was significantly increased in the abortion sites compared with the implantation sites of abortion model mice on day 14.5 and 19.5 of pregnancy. In addition, the expression levels of IL-17A mRNA were significantly higher in abortion sites than in implantation sites on day 14.5 and 19.5. Moreover, the nesfatin-1/NUCB2 protein and mRNA levels were increased in abortion sites compared with levels in implantation sites of both normal pregnant and abortion model mice on day 14.5 of pregnancy. Interestingly, nesfatin- 1/NUCB2 serum levels were not changed throughout the whole pregnancy in abortion model mice, but its serum level was dramatically increased on day 14.5, and then rapidly decreased on day 19.5 in normal pregnant mice. In this study, we showed for the first time the expression of nesfatin-1/NUCB2 mRNA and protein in implantation sites during pregnancy. The present results suggest that Th17 cells in the uterus may play an important role in the period of implantation and for maintenance of pregnancy. Furthermore, the present results suggest that Th17 cells in implantation sites may be a key regulator for maintenance of pregnancy and provides evidence that activation of these cells may be regulated by nesfatin-1/NUCB2. Further study is needed to elucidate the role of nesfatin-1 expressed in the uterus during pregnancy.

This study was examined the ovogenesis of Ussurian bullhead, Leiocassis ussuriensis and the morphological development of its larvae and juveniles and to use the results as basic information for the preservation of species and resource enhancement. For artificial egg collection, human chorionic gonadotropin (HCG) was injected at a rate of 10 IU per gram of fish weight. During breeding period, water temperature maintained at 24.5~26.5℃ (mean 25.0±0.05℃). The process of ovogenesis reached the two-cell stage in 50 minutes after fertilization. In 73 hours of fertilization the movement of the embryoid body became active state and the larvae began to hatch from the tail through the oolemma. Length of prelarvae were 6.33~6.50 mm long (mean 6.40±0.06 mm) just after hatching having yolk with their mouth not opened. After thirty eight days of hatching, juveniles were 30.6∼32.5 mm long (mean 31.5±0.65 mm). The color was dark yellowish brown throughout the entire body, and the number of caudal fin rays developed to thirty six perfectly.

Coronal Mass Ejections (CME), which originate from active regions of the Sun’s surface, e.g., sunspots, result in geomagnetic storms on Earth. The variation of the Earth’s geomagnetic field during such storms induces surface currents that could cause breakdowns in electricity power grids. Hence, it is essential to both monitor Geomagnetically Induced Currents (GICs) in real time and analyze previous GIC data. In 2012, in order to monitor the variation of GICs, the Korean Space Weather Center (KSWC) installed an induced current measurement system at SINGAPYEONG Substation, which is equipped with 765 kV extra-high-voltage transformers. Furthermore, in 2014, two induced current measurement systems were installed on the 345 kV high-voltage transformers at the MIGEUM and SINPOCHEON substations. This paper reports the installation process of the induced current measurement systems at these three substations. Furthermore, it presents the results of both an analysis performed using GIC data measured at the SINGAPYEONG Substation during periods of geomagnetic storms from July 2013 through April 2015 and the comparison between the obtained GIC data and magnetic field variation (dH/dt) data measured at the Icheon geomagnetic observatory.

Several optical monitoring strategies by a ground-based telescope to protect a Geostationary Earth Orbit (GEO) satellite from collisions with close approaching objects were investigated. Geostationary Transfer Orbit (GTO) objects, Inclined GeoSynchronous Orbit (IGSO) objects, and drifted GEO objects forced by natural perturbations are hazardous to operational GEO satellites regarding issues related to close approaches. The status of these objects was analyzed on the basis of their orbital characteristics in Two-Line Element (TLE) data from the Joint Space Operation Center (JSpOC). We confirmed the conjunction probability with all catalogued objects for the domestic operational GEO satellite, Communication, Ocean and Meteorological Satellite (COMS) using the Conjunction Analysis Tools by Analytical Graphics, Inc (AGI). The longitudinal drift rates of GeoSynchronous Orbit (GSO) objects were calculated, with an analytic method and they were confirmed using the Systems Tool Kit by AGI. The required monitoring area was determined from the expected drift duration and inclination of the simulated target. The optical monitoring strategy for the target area was analyzed through the orbit determination accuracy. For this purpose, the close approach of Russian satellite Raduga 1-7 to Korean COMS in 2011 was selected.

By using the Optical Wide-field Patrol (OWL) network developed by the Korea Astronomy and Space Science Institute (KASI) we generated the right ascension and declination angle data from optical observation of Low Earth Orbit (LEO) satellites. We performed an analysis to verify the optimum number of observations needed per arc for successful estimation of orbit. The currently functioning OWL observatories are located in Daejeon (South Korea), Songino (Mongolia), and Oukaïmeden (Morocco). The Daejeon Observatory is functioning as a test bed. In this study, the observed targets were Gravity Probe B, COSMOS 1455, COSMOS 1726, COSMOS 2428, SEASAT 1, ATV-5, and CryoSat-2 (all in LEO). These satellites were observed from the test bed and the Songino Observatory of the OWL network during 21 nights in 2014 and 2015. After we estimated the orbit from systematically selected sets of observation points (20, 50, 100, and 150) for each pass, we compared the difference between the orbit estimates for each case, and the Two Line Element set (TLE) from the Joint Space Operation Center (JSpOC). Then, we determined the average of the difference and selected the optimal observation points by comparing the average values.

As a governmentally approved domestic entity for Space Situational Awareness, Korea Astronomy and Space Science Institute (KASI) is developing and operating an optical telescopes system, Optical Wide-field PatroL (OWL) Network. During the test phase of this system, it is necessary to determine the range of brightness of the observable satellites. We have defined standard magnitude for Low Earth Orbit (LEO) satellites to calibrate their luminosity in terms of standard parameters such as distance, phase angle, and angular rate. In this work, we report the optical brightness range of five LEO Satellites using OWL-Net.

To overcome the risk of the ovarian hyperstimulation syndrome (OHSS) in patients have polycystic ovarian syndrome (PCOS) and to prepare emergency fertility preservation in patients undergoing anticancer treatment, several researchers have reported IVM of oocytes retrieved from ovaries exposed by only hCG priming. However, the maturation rate and the developmental potential of embryos from IVM oocytes are significantly lower than those of oocytes matured in vivo. Here, we investigated the optimal time point for immature oocyte collection at post hCG only injection for in vitro maturation, in vitro fertilization and blastocyst formation. Immature GV oocytes were collected from 25 days old B6D2F1 female mouse at 12 hr, 14 hr, 16 hr or 24 hr post hCG injection. Oocytes were collected from antral or late secondary follicle by puncturing with 26 G needle. Collected oocytes were cultured in G2 medium with 10% FBS, FSH, estradiol, and hCG for 16 hr in vitro and subjected in vitro fertilization and further embryonic development. To examine follicular maturation, we estimated the numbers of primordial, primary, secondary follicle and antral follicle on ovaries of each time point post hCG. To confirm the optimal time point post hCG injection for collecting immature oocytes, we recovered the oocytes from each time point. There is no difference in the number of oocytes per mice. Oocytes collected at 14 hr post hCG injection were shown higher maturation rate to MII stage and blastocyst formation compare to other three groups (p<0.01). However, there is no difference in the maturation rate on the other three groups. Also, apoptotic signal with TUNEL assay or anti-PARP staining was not change in ovaries from all experimental groups. Granulosa cell proliferation test with anti Ki-67 or anti AMH was not show any difference. According to these results, there are no significant differences in four different time points at 12 hr, 14 hr, 16 hr or 24 hr of collection of immature oocytes in hCG primed mouse. However, oocytes from 14 hr post hCG injection showed higher percentages of maturation rate, in vitro fertilization rate, blastocyst formation.

Endocrine disruptors are exogenous chemicals that their endocrine disrupting effects mediated by androgenic signaling plays crucial roles in the control of development and several androgen-related diseases. However, there are no authorized in vitro screening and testing methods to evaluation of (anti-)androgenic activity. To find out a better in vitro cell line model, we have previously reported that 22Rv1 cells, a human prostate cancer cells contained functional Androgen Receptor (AR), might be an appropriate model for the evaluation of (anti-)androgenic endocrine disruptors. Based on this result, we developed a stable 22Rv1/mouse mammary tumor virus (MMTV) cell line to test AR-mediated transcriptional activation (TA). Using 22Rv1/MMTV cells, we established the test protocol and optimized the testing condition for AR-TA assay. In this study, we performed the inter-validation assay by four different laboratories to evaluate the 20 coded chemicals which were selected from the ICCVAM list (ICCVAM, 2003) or academic articles that exhibited exact (anti-) androgenic activity. The statistical analysis of the results of the inter-laboratory validation study revealed that there was reproducibility between the four participating laboratories. In conclusion, 22Rv1/MMTV AR-TA assay might be a quick and relatively inexpensive method, which can be used to screen large numbers of chemicals for their potential to activate or inhibit AR-mediated gene transcription. Furthermore, it will provide mechanistic data relevant to understanding adverse reactions observed in intact organisms.

Cryopreservation has become a powerful method of the assisted reproduction technology and supports fertility preservation of cancer and other indication patients. After controlled ovarian hyperstimulation, surplus oocytes and embryos were recommended to store using cryopreservation. Recently, vitrification is replaced with traditional slow freezing protocol, because of improved survival rates and clinical outcomes. Vitrification requires a high concentration of CPAs that may induce significant osmotic and metabolic damage to cells including oocytes even in a short exposure of a few minutes. Generally, MPF plays a crucial role in the cell cycle regulation and maintaining the meiotic arrest of oocytes. In fact, it has been observed to decline in MII ovine oocytes after vitrification and would be suggested that one of the main causes of low fertilization rate and developmental competence derived from cryoinjury during vitrification. Therefore, the aim of this study was to evaluate the effect of caffeine treatment on the activity of MPF, MAPK level in vitrified/warmed mouse mature eggs. Caffeine, Phosphataseinhibitor, may maintain active form of MPF. We evaluated their survival after warming procedure, fertilization, cleavage, and developmental rates. Ovulated MII eggs were retrieved from 6 weeks old B6D2F1 female mouse at 14hr post hCG injection. Collected MII eggs were maintained in HTF medium containing 10% KSR with or without caffeine for 1hr. Eggs were vitrified in 7.5%EG +7.5%DMSO equilibrium solution, 15%EG + 15%DMSO + 0.5M sucrose vitrification solution with or without caffeine. Also warming solution contained sucrose (0.5M, 0.25M, 0.125M, and 0M) with or without caffeine. After warming, eggs were cultured in HTF medium with or without caffeine for 2 hr then fertilized with epididymal sperm in vitro and cultured in KSOM for 5 days to analyze embryonic development. Survival rates were similar in all experimental groups. However, fertilization rate was higher in with caffeine group compare to without caffeine significantly (80% vs. 85%, p<0.05). 2-cell and blastocyst formation were increased in caffeine group (p<0.05). MPF activity and MAP kinase activity were recovered in with caffeine group after vitrification/warming process. In conclusion, Caffeine may maintain MPF and MAPK level in vitrified/warmed MII eggs, and enhance fertilization and further embryonic development.

A strategy is needed for a regional survey of geosynchronous orbits (GSOs) to monitor known space objects and detect uncataloged space objects. On the basis of the Inter-Agency Debris Committee’s recommendation regarding the protected region of geosynchronous Earth orbit (GEO), target satellites with perigee and apogee of GEO ± 200 km and various inclinations are selected for analysis. The status of the GSO region was analyzed using the satellite distribution based on the orbital characteristics in publicly available two-line element data. Natural perturbation effects cause inactive satellites to drift to two stable longitudinal points. Active satellites usually maintain the designed positions as a result of regular or irregular maneuver operations against their natural drift. To analyze the detection rate of a single optical telescope, 152 out of 412 active satellites and 135 out of 288 inactive satellites in the GSO region were selected on the basis of their visibility at the observation site in Daejeon, Korea. By using various vertical view ranges and various numbers of observations of the GSO region, the detection efficiencies were analyzed for a single night, and the numbers of follow-up observations were determined. The orbital estimation accuracies were also checked using the arc length and number of observed data points to maintain the GSO satellite catalog.

We estimated the orbit of the Communication, Ocean and Meteorological Satellite (COMS), a Geostationary Earth Orbit (GEO) satellite, through data from actual optical observations using telescopes at the Sobaeksan Optical Astronomy Observatory (SOAO) of the Korea Astronomy and Space Science Institute (KASI), Optical Wide field Patrol (OWL) at KASI, and the Chungbuk National University Observatory (CNUO) from August 1, 2014, to January 13, 2015. The astrometric data of the satellite were extracted from the World Coordinate System (WCS) in the obtained images, and geometrically distorted errors were corrected. To handle the optically observed data, corrections were made for the observation time, light-travel time delay, shutter speed delay, and aberration. For final product, the sequential filter within the Orbit Determination Tool Kit (ODTK) was used for orbit estimation based on the results of optical observation. In addition, a comparative analysis was conducted between the precise orbit from the ephemeris of the COMS maintained by the satellite operator and the results of orbit estimation using optical observation. The orbits estimated in simulation agree with those estimated with actual optical observation data. The error in the results using optical observation data decreased with increasing number of observatories. Our results are useful for optimizing observation data for orbit estimation.

An algorithm to automatically extract coordinate and time information from optical observation data of geostationary orbit satellites (GEO satellites) or geosynchronous orbit satellites (GOS satellites) is developed. The optical wide-field patrol system is capable of automatic observation using a pre-arranged schedule. Therefore, if this type of automatic analysis algorithm is available, daily unmanned monitoring of GEO satellites can be possible. For data acquisition for development, the COMS1 satellite was observed with 1-s exposure time and 1-m interval. The images were grouped and processed in terms of “action”, and each action was composed of six or nine successive images. First, a reference image with the best quality in one action was selected. Next, the rest of the images in the action were geometrically transformed to fit in the horizontal coordinate system (expressed in azimuthal angle and elevation) of the reference image. Then, these images were median-combined to retain only the possible non-moving GEO candidates. By reverting the coordinate transformation of the positions of these GEO satellite candidates, the final coordinates could be calculated.

Storm sudden commencements (SSCs) occur due to a rapid compression of the Earth's magnetic field. This is generally believed to be caused by interplanetary (IP) shocks, but with exceptions. In this paper we explore possible causes of SSCs other than IP shocks through a statistical study of geomagnetic storms using SYM-H data provided by the World Data Center for Geomagnetism – Kyoto and by applying a superposed epoch analysis to simultaneous solar wind parameters obtained with the Advanced Composition Explorer (ACE) satellite. We select a total of 274 geomagnetic storms with minimum SYM-H of less than –30nT during 1998-2008 and regard them as SSCs if SYM-H increases by more than 10 nT over 10 minutes. Under this criterion, we found 103 geomagnetic storms with both SSC and IP shocks and 28 storms with SSC not associated with IP shocks. Storms in the former group share the property that the strength of the interplanetary magnetic field (IMF), proton density and proton velocity increase together with SYM-H, implying the action of IP shocks. During the storms in the latter group, only the proton density rises with SYM-H. We find that the density increase is associated with either high speed streams (HSSs) or interplanetary coronal mass ejections (ICMEs), and suggest that HSSs and ICMEs may be alternative contributors to SSCs.