뻐ny studìes have shown the anti-proli ferative effects of irondeprivation on cancer cell s‘ but the effects 01' iron-chelators on oral cancer have not been clearly elucidated , To investigate the effects of an iron chelato r, desferrioxamine( 01"O).on the growth of ilIllTIortali zed human o1'al ke ratinocytes(IHOK), primary oraJ cancer cel ls(HN4)‘ metastatic oral cancer cell s(HN12) , and human skin keratìnocytes(HaCaT) in the MTr assay, three-dimensional(3D) raft cul tmes, Western blott ing, cell cycJ e analysis‘ nuclear staining‘ and cytochrome c expression for apoptosis s ig naling pathway were used OFO inhibited the growth of immortalized IHOK and HaCaT and mal ignant HN4 and HN12 keratinocytes in a time- and dose-dependent manner according to the MTT assay, The 3D organotypic cu l tu re also revealed that OF'O-treated cells showed less epithelial maturation, less surface keratinizati on‘ and de creased epithelia l thickness, The major mechanìsm of growth inhìbition with the micromolar 0 1"0 treatment was by the induction of apoptosis‘ which was supported by nuclear OAPI staining, ONA fragmentation analysis, and J10w cytometric analysis for sub-Gl phase ar rest and Annexin V-1"ITC stainìng, Furthermore‘ Bax expression in creased together with p53 and p21WAF1!CIPl, whìle the Bcl-2 expression decreased in the immortalized and malig nant keratinocytes treated with 01"0 , Time-dependent cytochrome c from mitochondria was observed in D1"O-treated [l-IOK and 0 1'머 cancel‘ ceJJ s, and was accompanied by the activation of caspase-3 in IHOK cells. These resu lts demonstrate that 0 1"0 has growth inhibitory effects on immortalized and malignant oral keratinocyLes Lhrough the induction of apoptosis and suggest that further evaluation of OFO as a potcntial thcrapcutic agent for human oral precancerous and cancerous lesions is warranted

Radish (Raphanus sativus L.) is a species of the Brassicaceae family and an important root vegetable crop, produced worldwide. A total of 82 radish accessions with various morphological and physiological characteristics analyzed for total sugars content. These accessions includes five subspecies and classified as wild, wild-relative, traditional and improved cultivar. The four sugars, glucose, fructose, sucrose and maltose, showed various contents in 82 accessions. Total sugar content ranged from 5.64 to 46.68 ㎎/g and showed average 25.33 ㎎/g. Total sugar content was not statistically significant among the five subspecies, but individual sugar ratio varied. The wild, wild-relatives and traditional cultivars were not significantly among average total sugars content compared with improved cultivars. On the other hand, the wild and traditional cultivars were showed high ratio of individual sugars. These results could be valuable information for the development of new radish cultivars and regulation of sugars biosynthesis in radish.

Dual-color fluorescence in situ hybridization karyotype analysis was created using repetitive sequences including two types of rDNA repeats (45S and 5S rDNAs) and Arabidopsis-type telomere sequence repeats. The somatic metaphase cells of Carthamus tinctorius were observed as diploids (2n=2x=24). A symmetrical or slightly asymmetrical karyotype with seven pairs of metacentric and five pairs of submetacentric chromosomes was observed. The lengths of the somatic metaphase chromosomes ranged from 4.18 to 6.53 ㎛, with a total length of 60.71 ㎛. One locus of 45S rDNA was located on the pericentromeric regions of three pairs of chromosomes and the other pair was situated on the terminal regions of the short arms of a single pair of chromosomes. One locus of 5S rDNA was detected on the interstitial regions of the short arms of two pairs of chromosomes. Arabidopsis-type telomeric repeats were detected on the terminal regions of all pairs of chromosomes. Co-localization of loci between telomeric repeats and 45S rDNA was observed in a single pair of chromosomes. The results provide additional information for the existing physical mapping project of C. tinctorius and will also serve as a benchmark to a more intricate cytogenetic investigation of C. tinctorius and its related species.

The Asteraceae/Compositae family is one of the biggest families in flowering plants and has more than 23000 species including the economically important lettuce, sunflower, and chicory as well as the agronomic weeds. With its significance and the progress in sequencing technology, its species have been subjected to the genome sequencing project worldwide. Although chrysanthemum is an important plant in the floricultural industry, however, it has been less studied at the level of genomics, compared with other species in the Asteraceae. There were only several reports on comparative analysis of transcriptome for chrysanthemum. Actually, the genome of Chrysanthemum species is known to be gigantic and complex with diverse status ranging from diploid to decaploid. Since the cultivated and commercial chrysanthemum exhibits hexaploid genome, we decided to select the diploid species with smaller genome as a material for reference genome sequencing. Thus, we launched a genome sequencing project with C. boreale which was previously reported to be diploid by cytogenetic analysis. We constructed sequencing libraries with insert size 300bp and 500bp and sequenced them from the paired end in 100bp read length with Illumina’s HiSeq platform. After quality checking, we preprocessed raw reads by removing duplicated reads and trimming reads with low quality value. Kmer frequency analysis with the cleaned reads showed that the genome is heterozygous, highly repetitive and gigantic, ranging from 2.9Gb to 5.8Gb. The cleaned reads were further subjected to error correction and primary assembly with SOAPdenovo2. Here, we’ll report the result of Kmer frequency analysis and genome assembly.

In Brassica as matter of seedling manner, they have the bilocular ovary and 20~28 seeds per silique after fertilization. Rarely some of B. juncea and yellow sarson (Brassica rapa ssp, tricolaris) have multilocular ovary. In this stdudy, the LP8 (YS-033, CGN06835) is shown tetralocular ovary as well as high seed yields. As microscope study for the different size of immature bud sections and we have known the floral meristem with already four locules in immature buds less size than 1mm of LP8. To identify of determining of tetralocular ovary formation, RNA-seq was carried out on the isolated RNA from less than 1mm and from 1mm of bud size respectively. By contrast tetralocular ovay and bilocular ovary, Chiifu is used. A total of 994 differentially expressed genes(DEGs) are detected in only LP8. Among the DEGs, we identify 18 DEGs in only immature buds of less size than 1mm. The expression patterns of 18 DEGs are validated by real time quantitative PCR and these genes are cloned and the sequence analyzed. At present, 12 candidated gene are analyzed by sequencing and there are detected by large fragment insertion as well as SNPs in sequence comparison to Chiifu. We will perform the genetic transformation of these DEG genes in Arabidopsis for relation between genes and tetralocular ovary. Our results will be helpful in understanding for mechanisms of tetraovular ovary in Brassica rapa.

Brassica rapa subspecies show morphological variability, containing vegetable types and oilseed types. The yellow sarson types(Brassica rapa ssp, tricolaris) have distinct morphology, yellow seeded and contain some lines with very unique character of tetralocular ovary. For genetic studies on tetralocular ovary related to high seed yields, we produced genetic segregation population with F2 and double haploid(DH) population. The yellow sarson LP8 (YS-033, CGN06835) with character of tetralocular ovary used as a maternal plant and crossed by LP21 of turnip rape type with bilocular ovary as paternal plant. We took on the microspore cultures on immature bud which is collected on sizing from 2mm to 3.2mm for DH population. The regenerations DH plants are analyzed by ploidy determination using flow cytrometer and selected on diploid plants. These regenerated DH and F2 plants are doing bud pollination and measuring the phenotype traits. Also, these populations will be used for identify of genetic locus relate to tetralocular ovary using genotyping by sequencing.

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.

Glucosinolates of Brassica rapa collection from Korea genebank were measured to determine total glucosinolate content and their variation of diverse glucosinolates; Around 100 accessions representing the different morphotypes and geographical origin of Brassica rapa were analysed. The principal component analysis was performed to evaluate the differences among morphotypes using the profiles of 14 glucosinolates identified from the leaves. DMRT test and box plots showed the significant difference between total glucosinolates of subspecies. Most of turnip accessions had higher gluconilates compared to the other type accessions, Chinese cabbage and pak choi. These accessions will be used for GWAS study for glucosinolate. Now they are being finger-printed by genotyping by sequencing (GBS). Among these accession, we selected a turnip accession with high amount of glucosinolate, K0466 and two Chinese cabbage accession with low amount of glucosinolate, K0015 and K0621. To analyse quantitative traits loci (QTL) for glucosinolate synthesis, these three accessions were fixed through microspore culture. Finally, six homozygous lines were selected and were crossed each other to make F1 hybrids. We just harvested F2 seeds and transferred doubled haploid plants to pots. QTL analysis for glucosinolate will be performed these F2 and DH population.

This study was carried out to investigate the correlation between root diameter and ginsenoside compositionof Panax ginseng C. A. Meyer cultivar Yunpoong. Dry matter ratio of main root was a little higher than that of lateral rootand fine root, and that was higher by the increase of root diameter in the same root parts. Total ginsenosides composition ofmain and lateral roots increased by the decrease of root diameter, especially in lateral root. Similar resulted in fine root, butthere was no significant difference where root diameter was below 2.5㎜. Except for ginsenoside-Rg1, other ginsenosidescomponent, PDs, PTs and total ginsenosides had highly negative correlation with the root diameter within whole root, mainroot+lateral root and lateral root+fine root, while Rg1 had positive correlation with the root diameter.