Background : Due to immature development of embryo in ripened berries, dehiscence process is required for the proper germination of ginseng seeds. Such process involves the preparation of the container with alternating layers of seed and moist sand. In order to make sand fully moist, water sprayer has been usually used by farmers, which is labor intensive, time consuming and causing uneven sand moisture. Methods and Results : In this study, we investigated the effects of different stratification methods on dehiscence ratio of ginseng seeds. Ginseng seeds were stratified for 90 days in a total of 12 different treatments and the dehiscence ratios were compared; drainage methods, drainage time, the ratio between ginseng seeds and sand, and etc. Seed stratification process was performed according to the guideline of ginseng GAP. One thousand ginseng seeds were used for each treatment. It was found that the average of dehiscence of the 12 treatments was 84.6 %. The highest dehiscence ratio (90.3 %) was observed in the seeds that were treated with water soaking, immediately followed by drainage. Higher ratio was also observed in the seeds that were soaked for 60 min, followed by drainage. Therefore, our findings indicate that ginseng seeds soaked in water less than 60 min could dehisce more efficiently than traditional method. Conclusion : Our study demonstrated that ginseng seeds that are subjected to water soaking and then drainage showed better ration of dehiscence. This method will eventually decrease the time and labor used for seed stratification.

Background : This study was conducted to determine the impact of temperature elevated and the effect of transplanting times based on climate change scenario on growth of 2-year-old korean ginseng (Panax ginseng C. A. Meyer.) in temperature gradient chambers (TGC). Methods and Results : As a plant materials, ‘Yunpoong’ was cultivated in TGC at ambient temperature(Amb), Amb+2℃, Amb+4℃ and Amb+6℃ respectively. Ginseng was also transplanted on March 29, April 12 and 26 respectively. Investigation on characteristic of aerial parts were carried out on 28, 56, 84 and 112 days after transplanting and characteristic of roots were conducted on October 19. As transplanting time was faster and temperature was higher, the growth of aerial parts were increased. Compared with those of ginseng transplanted on March 29 with Amb, the root weight which tend to decrease depending on late transplanting time and high temperature decreased about 11.1%, 35.4% and 42.4% in Amb+ 2℃, Amb+4℃ and Amb+6℃ respectively. Ginseng transplanted on April 12 and 26 decreased about 20.9%, 33.9% respectively. Conclusion : Consequently, the more transplanting time extend, the more quantity increased in all temperature treatment. So, it is possible to increase in quantity to advance transplanting time although high temperature will be caused by the climate change.

Background : When ginseng seeds were gathered, the seeds were unripe. To grow immature embryo definitely, special treatment called dehiscence must be performed. Even though dehiscence is completed, most ginseng seeds are on enforced dormancy. The breaking seed dormancy is generally achieved using cold treatment. Also it is reported that gibberellin treatment can replace the treatment. It is very time consuming process in order to develop new ginseng cultivar because ginseng flowers after 3 years of growth. To shorten the ginseng breeding period, it is necessary to establish fast generation progress. Therefore, this study examined the possibility of breaking seed dormancy of ginseng using GA3 treatment and alternating temperature. Methods and Results : Seeds were obtained from local variety fruit which is not inbred. Gibberellin of 100 ppm was treated at seeds for 24 hours. Fixed cold condition was treated on both –2℃ and 2℃. Alternating cold condition was treated on 2℃ and then –2℃, finally 2℃. Fixed and alternating temperature was continued for 15, 30, 45, 60, 90 days that 15 days of alternating temperature is first 2℃ for 5days and then -2℃ for 5days, finally 2℃ for 5days. The other treatment periods such as 30, 45, 60, 90 days mean 10, 15, 20, 30 days respectively. Each of 48 seeds were sowed on tray in greenhouse at 3 replication. Experimental plot was completely randomized. Conclusion : Seeds untreated with GA3 were germinated little and there is no difference between 2℃ and –2℃. Alternating temperature until 60days made no difference with fixed temperature but germination rate increased up to 70.8% when seeds were treated for 90days. Germination of seeds treated with GA3 is much higher than untreated seeds especially combined with alternating temperature.

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.

To understand molecular mechanisms underlying adaptation of plant cells to saline stress and stress memory, we developed Arabidopsis callus suspension-cultured cells adapted to high salt. Adapted cells to high salt exhibited enhanced tolerance compared to control cells. Moreover, the salt tolerance of adapted cells was stably maintained even after the stress is relieved, indicating that the salt tolerance of adapted cells was memorized. Salt-adapted and stress memorized cells were densely aggregated and formed multi-layered cell lump. Cell morphology analysis using transmission electron microscopy indicated that cell wall thickness of salt-adapted cells was significantly induced compared to control cells. In order to characterize metabolic responses of plant cells during adaptation to high salt stress as well as stress memory, we compared metabolic profiles of salt-adapted and stress-memorized cells with control cells by using NMR spectroscopy. A principle component analysis showed clear metabolic discrimination among control, salt-adapted and stress-memorized cells. Compared with control cells, metabolites related to shikimate metabolism such as tyrosine, and flavonol glycosides, which are related to protective mechanism of plant against stresses were largely up-regulated in adapted cell lines. Moreover, coniferin, a precursor of lignin, was more abundant in salt-adapted cells than control cells. The results provide new insight into metabolic level mechanisms of plant adaptation to saline stress as well as stress memory.

To identify novel signaling components involved in regulation of plant responses to phosphate (Pi) starvation, we screened an Arabidopsis T-DNA activation tagging library for mutants with altered Pi-starvation responses. Here, we report the identification and characterization of novel activation-tagged mutant involved in Pi starvation signaling in Arabidopsis. The hpd (hypersensitive to Pi deficiency) mutant exhibits enhanced phosphate uptake and altered root architectural change under Pi starvation compared to wild type. Expression analysis of auxin-responsive DR5::GUS reporter gene in hpd mutant indicated that both auxin biosynthesis and auxin translocation under Pi starvation are suppressed in hpd mutant plants. Impaired auxin translocation in roots of hpd mutant was attributable to abnormal root architecture changes in Pi starvation conditions. Mis-regulation of auxin translocation in hpd mutant was further confirmed by analysis of expression patterns of auxin efflux carrier proteins, PIN-FORMED (PIN) 1, 2, and 3 fused with GFP. Not only expression levels but also expression domains of PIN proteins were altered in hpd mutant in response to Pi starvation. Molecular genetic analysis of hpd mutant revealed that the mutant phenotype is caused by the lesion in ENHANCED SILENCING PHENOTYPE4 (ESP4) gene whose function is proposed in mRNA 3’-end processing. The results propose that mRNA processing plays crucial roles in Pi homeostasis as well as developmental reprograming in response to Pi deprivation in Arabidopsis.

There are many known renal complications of rheumatoid arthritis. These complications can develop by the extraarticular aspect of rheumatoid arthritis or the use of disease modifying antirheumatic drugs or pain-killer. The most common renal disease induced by use of disease modifying antirheumatic drugs is membranous nephropathy. We report on a case of bucillamine induced nephropathy with prominent proteinuria and hypoalbuminemia. The renal biopsy result was stage I to II membranous nephropathy. After withdrawal of bucillamine the proteinuria improved and hypoalbuminemia recovered. Even in cases of accompanying prominent proteinuria and hypoalbuminemia, bucillamine nephropathy can be treated with withdrawal of bucillamine and immunosuppressant drugs are not required.

We determined intravitreal levels of vascular endothelial growth factor (VEGF) and pigment epithelium derived factor (PEDF) in patients with early phase rhegmatogenous retinal detachment (RRD). Twenty five eyes of 25 RRD patients with symptom onset from one day to 21 days prior to vitrectomy were selected. Concentrations of VEGF and PEDF in vitreous were determined by enzyme-linked immunosorbent assay and relationships between these concentrations and durations and involved quadrants were analyzed. Duration of RRD was found to show significant correlation with PEDF concentration. However, number of involved quadrants did not show correlation with PEDF concentration in vitreous.

The aim of this study was to analyze the effect of antifreeze proteins (AFPs) on vitrification of mouse mature (MII) oocytes. We studied about 3 types of AFPs from different origins (FfIBP, LeIBPand Type III AFP). The MII oocytes were obtained from 4-week-old BD-F1 mice. Vitrification of oocyte was performed by 2 steps using the Cryotop (equilibration: 7.5% EG + 7.5% PROH for 5 min, vitrification: 15% EG + 15% PROH + 0.5M sucrose for 1 min). The concentrations of AFPs added to these solutions were 0.05 mg/ml for FfIBP and 0.1 mg/ml for LeIBP and Type III AFP. After fertilization, embryo development was assessed up to 5 days. Through immunostaining of vitrified-warmed oocytes, we assessed the normal meiotic spindle. Also, intracellular ROS and mitochondrial activity was analyzed. In the developmental stages, FfIBP and LeIBP groups showed significantly higher survival rates. In the blastocyst and apoptotic blastomere rates were significant differences in AFPs treated groups. AFPs treated groups were significantly higher in blastocyst cell numbers than control group. Among the AFPs treated groups, FfIBP, LeIBP groups were significantly higher rates. And, in cleavage rates, FfIBP group was significantly higher rates than the other groups. In vitrified-warmed MII oocytes, the normal meiotic spindle organization and chromosome alignment rate was significantly higher in FfIBP and LeIBP groups. And in intracellular ROS levels, control group was significantly increased than AFPs treated groups. However, in the mitochondrial activity, LeIBP group was significantly higher than control, FfIBP and LeIBP groups. AFPs treated groups were significant differences in development, meiotic spindle organization and intracellular ROS levels. And in the AFPs treated groups, FfIBP and LeIBP groups were significantly higher rates in normal meiotic spindle and mitochondrial activity than Type III AFP group respectively. In conclusion, FfIBP and LeIBP can be thought to improve oocyte cryopreservation efficiency.

The follicle loss of transplanted ovarian tissue (OT) is caused by ischemia and slow revascularization. To shorten the ischemic period and promote angiogenesis, some angiogenic factors have been treated for transplanted tissues. Angiopoietin-2 (ANG-2) is one of the major angiogenic factors and has been reported to promote blood vessels and increase vascular permeability in the ischemic and/or hypoxic environment. So, this study was designed to assess the impact of ANG-2 on follicle integrity and revascularization of mouse OT grafts. The 5-week-old B6D2F1 female mice were divided into 3 groups (a control and 2 ANG-2 groups) followed by ovary collection and vitrification. After warming, the ovaries were autotransplanted into kidney capsules with/without ANG-2 injection (50 or 500 ng/kg), and then killed at day(D)2, 7, 21 and 42 after transplantation. Total 2,437 follicles in OT grafts were assessed for the follicular density, integrity and classification by H&E staining. Apoptosis, revascularization, and serum FSH levels were evaluated by TUNEL assay, CD31 immunohistochemistry, and ELISA respectively. All the ANG-2 groups showed remarkable increase of morphologically intact follicle ratio across all the grafting duration except D21 (no statistical difference). The numbers of CD31(+) vessels (the sum of 3 fields at ×400 magnification) were significantly increased in both ANG-2 groups compared with the control group at all the grafting duration. Especially at D42, the 500ng ANG-2 group showed significantly more vessels than the 50 ng ANG-2 group as well as the control group. However the mean follicle numbers of grafts, apoptosis ratio and serum FSH levels showed no significant difference among the groups. In this study, remarkably well preserved follicles and larger amount of vessels were appeared in ANG-2 treated groups. So we thought that ANG-2 treatment is effective for OT transplantation and improve transplantation outcomes.

Polyethyleneglycol-adsorbed–superoxide dismutase (PEG-SOD), has been proposed as an effective agent for reducing free radical-mediated injury. The objective of this study was to investigate a protective effect of PEG-SOD supplementation on ovarian tissue during transplantation. Ovaries from F-1 mice were collected and vitrified. After warming, ovaries were autotransplanted under kidney capsule. Mice were randomly divided into four groups according to dose of PEG-SOD, (0 U/ml, 100 U/ml, 1,000 U/ml and 10,000 U/ml respectively). Grafted ovaries were retrieved 2, 7 and 21 days later. PEG-SOD was treated by intraperitoneal injection once every 48 hours and especially for 21 days group, after first week treatment, PEG-SOD was treated once every 4 days. Morphology of ovaries was assessed histological analysis and ELISA for FSH was performed to evaluate restoration of ovarian function. In 2 days groups, morphologically intact follicle ratio of 10,000 U/ml group was significantly higher than other groups. In 7 days groups, morphologically intact follicle ratio was significantly higher in all treatment groups. In 21 days groups, there was no significant difference of intact follicle ratio in total follicles in all groups but intact primordial, primary and secondary follicles ratio was higher in 10,000 U/ml group. FSH levels in blood serum were decrease as time goes on, but there is no statistical difference in each groups. In conclusion, the data of the present study show that PEG-SOD has a beneficial effect on preservation of the morphologically intact follicle.

Ovarian tissue cryopreservation and transplantation causes follicle depletion. To overcome this problem, we investigate the effect of Anti-Müllerian hormone (AMH), a follicle recruitment control hormone, supplementation before and/or after mouse ovarian transplantation. A total of 120 5-week-aged BD F-1 female mice were used. The mice were randomly divided into four groups according to AMH doses (0, 5, 25, 125 μg/mL, respectively). AMH was injected intraperitoneally on every other day for a week before, after, or before and after transplantation of ovaries under kidney capsules was performed. One week after transplantation, follicular normality was evaluated by histological analysis and TUNEL assay. In Group A and C, morphologically intact follicle (G1) ratios of AMH treated groups showed no statistically significant difference. In Group B, G1 ratios of 25 and 125 μg/mL of AMH treated groups were higher than those of 5 μg/mL treated group, but there was no improvement in G1 ratio after AMH treatment. In every group, apoptotic follicle ratios did not show any trend according to AMH treatment. Proportions of primordial follicle were not significantly different according to AMH treatment in all groups. The result of the present study demonstrated that AMH treatment during on transplantation of cryopreserved ovaries has no significant effect on follicle survival and prevention of follicle depletion.

Objective : To investigate the effects of Simvastatin and Methylprednisolone on ovarian tissue cryopreservation and transplantation using mouse models. Methods : The mice were randomly distributed into 1 control and 3 experimental groups. The B6D2F1 mice were given oral Simvastatin (5 mg/kg), intravenous Methylprednisolone (15 mg/kg), or a combination of both at 2 hours before ovariectomy. Same volume of normal saline was given perorally in the control group at 2 hours before ovariectomy. The ovarian tissues were vitrified accrording to our protocols. The vitrified ovaries were warmed 1 week later and auto-transplanted under bilateral kidney capsules. The ovaries and blood sera were collected at 2, 7 or 21 days after transplantation. Histological analysis, TUNEL assay, immuno-histochemistry for CD31, serum AMH level and embryonic development after in vitro fertilization were assessed for evaluation. Results : With regard to the total grade 1 follicle rate, both Simvastatin or Methylprednisolone treated groups were significantly increased at 2, 7 or 21 days after transplantation (except Simvastatin treated group at 7 days). A combination of Simvastatin and Methylprednisolone group was significantly improved in terms of the total G1 follicle rate, apoptotic follicle rate, CD31 positive area and serum AMH after ovarian tissue transplantation. However, there were no statistically difference with respect to the oocyte maturation rate, blastulation rate, and the other embryonic development parameters after in vitro fertilization procedure among the four groups. Conclusion : Our results suggest that combined donor Simvastatin and Methylprednisolone have beneficial effects on the quality and function of transplanted ovarian tissues.

Study question: What is the optimal vitrification protocol according to the cryoprotective agent (CPA) for ovarian tissue (OT) cryopreservation? Summary answer: The two-step protocol with 7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO) for 10 min then 20% EG, 20% DMSO and 0.5 M sucrose for 5 min showed the best results in mouse OT vitrification. What is known already: Establishing the optimal cryopreservation protocol is one of the most important steps to improve OT survival. However, only a few studies have compared vitrification protocols with different CPAs and investigated the effect of in vitro culture (IVC) on vitrified–.warmed OT survival. Some recent papers proposed that a combination of CPAs has less toxicity than one type of CPA. However, the efficacy of different types and concentrations of CPA are not yet well documented. Study design, size, duration: A total of 644 ovaries were collected from 4-week-old BDF1 mice, of which 571 ovaries were randomly assigned to 8 groups and vitrified using different protocols according to CPA composition and the remaining 73 ovaries were used as controls. After warming, each of the eight groups of ovaries was further randomly divided into four subgroups and in vitro cultured for 0, 0.5, 2 and 4 h, respectively. Ovaries of the best two groups among the eight groups were autotransplanted after IVC. Participants/materials, setting, methods: The CPA solutions for the eight groups were composed of EDS, ES, ED, EPS, EF, EFS, E and EP, respectively (E, EG; D, DMSO; P, propanediol; S, sucrose; F, Ficoll). The IVC medium was composed of a-minimal essential medium, 10% fetal bovine serum and 10 mIU/ml follicle-stimulating hormone (FSH). Autotransplantation of vitrified–.warmed OTs after IVC (0 to 4 h) using the EDS or ES protocol was performed, and the grafts were recovered after 3 weeks. Ovarian follicles were assessed for morphology, apoptosis, proliferation and FSH level. Main results and the role of chance: The percentages of the morphologically intact (G1) and apoptotic follicles in each group at 0, 0.5, 2 and 4 h of IVC were compared. For G1 follicles at 0 and 4 h of IVC, the EDS group showed the best results at 63.8 and 46.6%, respectively, whereas the EP group showed the worst results at 42.2 and 12.8%, respectively. The apoptotic follicle ratio was lowest in the EDS group at 0 h (8.1%) and 0.5 h (12.7%) of IVC. All of the eight groups showed significant decreases in G1 follicles and increases in apoptotic follicles as IVC duration progressed. After autotransplantation, the EDS 0 h group showed a significantly higher G1 percentage (84.9%) than did the other groups (42.4–.58.8%), while only the ES 4 h group showed a significant decrease in the number of proliferative cells (80.6%, 87.6–.92.9%). However, no significant differences in apoptotic rates and FSH levels were observed between the groups after autotransplantation. Limitations, reasons for caution: The limitation of this study was the absence of in vitro fertilization using oocytes obtained from OT grafts, which should be performed to confirm the outcomes of ovarian cryopreservation and transplantation. Wider implications of the findings: We compared eight vitrification protocols according to CPA composition and found the EDS protocol to be the optimal method among them. The data presented herein will help improve OT cryopreservation protocols for humans or other animals.

Genome sequencing researches for considerable numbers of crops and wild plants are being developed. Cytogenetic researches according to chromosome number and size are essential to confirm and comprehend ploidy level and genome size before genome sequencing project is actually conducted. Cytogenetic researches on six food crop plants were carried out by DAPI staining and fluorescence in situ hybridization (FISH) method. Fagopyrum esculentum Moench showed 2n=2x=16, each chromosome length of 1.42㎛ to 1.77㎛, total chromosome length of 13.31㎛, and karyotypic formula of 2n=8m; Phaseolus angularis W.F. Wight, 2n=2x=22, 2.01㎛ to 3.84㎛, total 28.03㎛, 2n=9m+2sm, Perilla frutescens var. japonica Hara, 2n=2x=40, 1.73㎛ to 2.76㎛, total 44.36㎛, 2n=5m+13sm+2st. Chromosome sizes of the other three species such as, Panicum miliaceum L., 2n=2x=36, total chromosome length of 30.83㎛, Sesamum indicum L., 2n=2x=26, 27.39㎛, lpomoea batatas L., 2n=2x=30, total 33.51㎛ were too small for each chromosome type to be identified and analyzed. The result of FISH analysis using 5S and 45S rDNA probe showed species-specific chromosome locations in the genome. These preliminary analyses were carried out to decide which food crop to prioritize for genome sequencing. This work was supported by the “Cooperative Research Program for Agriculture Science & Technology Development (No.PJ009837), Rural Development Administration, Republic of Korea.

본 연구는 삼채뿌리분말을 첨가하여 김치의 맛과 저장성 을 증진시키는 김치를 개발하고자 품질특성을 관찰하였다. 삼채뿌리분말 0.1%, 1%, 5%를 첨가하여 4℃에서 8주 동안 저장하였으며 저장기간에 따른 pH, 산도, 염도, 환원당, 미 생물의 변화 및 관능특성을 살펴보았다. 삼채뿌리의 비타 민 B1및 비타민 C의 함량은 각각 0.04 mg/100 g 및 5.76 mg/100 g으로 나타났으며, 삼채 뿌리의 비타민 B1의 함량은 마늘보다는 적고 양파와 비슷한 수준으로 검출되었다. 삼 채뿌리분말 첨가 김치의 pH의 측정결과 초기의 pH는 비슷 하였으나, 저장기간 중 삼채뿌리분말을 첨가하지 않은 일 반김치는 삼채뿌리분말을 첨가한 김치에 비하여 대체적으 로 낮은 pH를 보였으며, 삼채뿌리분말 5% 첨가한 김치의 pH가 가장 높게 나타났다. 산도의 측정결과 pH 변화와 비슷 하였으며, 삼채뿌리분말첨가 김치의 염도는 대체로 2.23% 를 유지하였다. 환원당 함량의 측정 결과 삼채뿌리분말 0.1%, 1%, 5% 첨가 김치는 발효가 진행됨에 따라 모든 군에서 환원당 함량이 지속적으로 감소되었고, 삼채뿌리분 말 첨가량이 많을수록 환원당 함량이 높은 것을 알 수 있었 다. 숙성 중 총 균수 및 젖산균수 변화는 숙성 초기에 급격히 증가한 후 서서히 증가하는 경향을 보였다. 관능적 특성을 살펴보면 외관, 맛, 전반적인 기호도 에서 삼채뿌리분말 1% 첨가 김치의 평가가 가장 우수하였다. 이상의 결과를 보면 삼채뿌리분말을 첨가한 김치의 pH가 높고 천천히 감 소되는 점으로 미루어 볼 때, 숙성 지연 등의 저장성과 기호 성이 좋은 삼채뿌리분말 1%를 첨가하여 김치를 개발하는 것이 적합한 것으로 사료된다.

In order to adapt to various environmental stresses, plants have employed diverse regulatory mechanisms of gene expression. Epigenetic changes, such as DNA methylation and histone modifications play an important role in gene expression regulation under stress condition. It has been known that some of epigenetic modifications are stably inherited after mitotic and meiotic cell divisions, which is known as stress memory. To understand molecular mechanisms underlying stress memory mediated by epigenetic modifications, we developed Arabidopsis suspension-cultured cell lines adapted to high salt by stepwise increases in the NaCl concentration up to 120 mM. Adapted cell line to 120 mM NaCl, named A120, exhibited enhanced salt tolerance compared to unadapted control cells (A0). Moreover, the salt tolerance of A120 cell line was stably maintained even in the absence of added NaCl, indicating that the salt tolerance of A120 cell line was memorized even after the stress is relieved. By using salt adapted and stress memorized cell lines, we intend to analyze the changes of DNA methylation, histone modification, transcriptome, and proteome to understand molecular mechanisms underlying stress adaptation as well as stress memory in plants.