1'0 determine Lhe ll1echanism of cell c1eath incluced by iron chelators. we explored the pathways of the three structurally relatecl ll1 itogen-activatecl protein(MAP) kinase subfami li esduring iron cbelator- inclucecl apoptosis ancl differentiation of oral precancerous ancl cancel‘ cells. The iron chelator c1 eferoxamine(DFO) exertecl potent timeancl c1ose-c1epenclent inhi bitory effects on the growth of IHOK and HN4 cells The major mechanism of growth inhibition following DFO treatment was fOllncl to be apoptosis incluction. as assessecl by annexin V-FITC staining. cell cycle analysis‘ DNA lacldering, a ncl Hoechst staining. We report that DF'O s trongly activates the p38 MAP kinase and extracell ular signal- regu lated kinase(ERK). but c10es not activate the c-Jun N-terminal kin ase/ stl않s-activaLecl protein kinase(JNK/8APK) . Of the three MAP kinase blockers usecl‘ the selective p38 MAP kinase inhibitor 8ß203580 ancl ERK inhibitor PD98059 protected oral premaIignant ancl malignant cells againsL iron chelator- nclllced cell death. which incl icates that the p38 MAP kinase serves as a major mecliator 01' apoptos is induced by this iron chelator DFO also evoked the release of cytochrome c from mitochondria, and incluced the activation of caspase-3 ancl caspase-8 in oral cancer cells, which suggests that apoptosis occurs via the mi tochoncl ri on - mecl iaLed pathway. DFO enhanced the expression of Bax in IHOK ancl HN4 cells. consistent witll thei r p53 status Moreover. DFO downregulatecl the expression 01' Bcl-2 in oral cancer cells. which suggests that DFO- incluced apoptos is 01' oraJ cancer and precancerous cells may be mediatecl by an increase in the ratio of pro-apoptotic to anti-apoptotic proteins. ln terestingJy, trcatmcnt 01' IHOK ancl HN4 cel ls with 8B203580 abolishecl cytochrome c release‘ as wel l as the activation of caspase-3 and caspase-8. DFO suppressecl the expression of epithelial di ffe rentiation markers, such as involucrin, t ransglutaminase II. CK6. and CK19. ancl this suppression was blockecl by p38 ancl ERK MAP kinase ll1hlbltors The oral premalignant(IHOK) ancl malignant cell s(I-lN4) showed differential responses to DFO with respect to the expression of cel l cycle regulatory proteins. cell growth. ancl apoptosis. Coll ectively. the current study reveals that p38 MAP kinase plays an ill1 portant role in iron chela tor-mecliatecl cel l cleath and in the suppression of differentiation of oral premalignantandmalignanLcell s.by activating a c10wnstream apoptotic cascade that executes the ceIl c1eath pathway

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

This study was carried out to examine the effects of NaOCl, sucrose, and BA concentration as pretreatment solution on quality and vase life of cut flowers in Dendranthema grandiflorum ‘Jinba’. Flower diameter, fresh weight, and vase life in control and 0 mg･L -1 NaOCl treatments decreased, and the treatment with 100~200 mg･L -1 NaOCl was more effective in the quality and vase life. In pretreatment with 2.5% sucrose solution, flower diameter and fresh weight decreased and vase life was shortest due to the rapid leaf wilting. However, pretreatment with 0.1% sucrose solution increased the flower diameter and fresh weight, and showed the longest vase life. When more than 80 mg･L -1 BA was treated with pretreatment solution, flower diameter and fresh weight decreased, and vase life was shortened. With pretreatment of 20 mg･L -1 BA, the flower diameter was bigger than in the other treatments, but it was no effect on fresh weight and vase life. Therefore, it was suggested that pretreatment solution mixed with 200 mg･L -1 NaOCl, 0.1% sucrose, and 20 mg･L -1 BA was the most effective for the quality and vase life of cut flowers in standard chrysanthemum ‘Jinba’.

Citrus canker caused by Xanthomonas citri is a notorious disease affecting a decrease in fruit productivity and quality. Citrus export to USA is also prohibited by the disease. Therefore, development of citrus canker resistant variety is essential and exploitation of markers for molecular breeding is urgent. To develop DNA molecular markers, we performed whole genome resequencing for 8 varieties: 4 citrus canker resistant varieties including C. hybrid ‘Kioymi’ and 4 citrus canker susceptible varieties including C. iyo ‘Miyauchiiyokan’. In total, 642 polymorphic SNPs were detected between resistant and susceptible varieties. Of the 642 SNPs, 50 SNPs were preferably selected based on integrative genomics viewer. To apply the markers in a broad range of citrus variety, we performed genotyping with 6 other varieties very well known as citrus canker resistant and susceptible varieties in addition to previous mentioned 8 varieties. Three of the 50 SNPs were identified as a marker to distinguish citrus canker resistant varieties from susceptible varieties. Secondly, we developed molecular markers to apply for F1 lines crossed by ‘Kiyomi’ and ‘Miyauchiiyokan’. Of the 50 SNPs, we identified 2 SNP markers to distinguish between F1 resistant and susceptible lines. One of them is a resistance gene that plays a role in plant defense mechanism. In this study, we developed 5 molecular marker candidates possible to apply for molecular breeding to develop citrus canker resistant variety. We are working on development of candidate markers related to citrus canker.

Livestock wastewater has high potential as one of energy resources because this wastewater is including high organic matter. Therefore the studies attempting to bio-gasification and bio-electricity generation using livestock wastewater is being tried. The pre-treatment system used in this study was the purpose to control the ammonia nitrogen in conjunction with the system and the microbial fuel cell. Because ammonia nitrogen is to inhibit the electricity generation efficiency of microbial fuel cell. These studies were to ascertain the effect of oxidants on the nitrogen removal in the pre-treatment system using catalyst and microbubbles to treat the ammonia nitrogen. The three kinds of oxidant such as air, oxygen (O2), and hydrogen peroxide (H2O2) were used to know the ammonia and nitrate nitrogen removal. This system was operated with four kinds of conditions. First method is O2 gas with 100 mL/min with 1ml of 30% H2O2 was supplied to the wastewater. A second method, the O2, with 400 and 1,000 mL/min was supplied through the flow meter before livestock wastewater was flow in the reactor. The last method, air was supplied 800 mL/min. The nitrate removal had no significant difference all conditions except the air supply. On the other hand, the ammonia and nitrate nitrogen removal when oxygen was supplied with 1000 mL O2/min was higher than that of the other conditions. The removal rate when air was supplied 800 mL/min was similar to the value in the supplied with 400 ml O2/min. This result showed that oxidant was important factor to improve the ammonia nitrogen removal rate.

Citrus canker caused by Xanthomonas citri pv. citri is one of economically important diseases in the citrus industry. The devastating bacterial disease results in unattractive quality and a significant reduction in fruit production. Citrus growers and industry in Korea has been struggling with the serious disease causing the prohibition of export market. Korea also became the top import market for oranges. The development of markers linked to citrus canker resistance is strongly needed. In this study, we investigated molecular markers between ‘Kiyomi’ (Citrus unshiu ｘ C. sinensis), a resistant cultivar, and Natsudaidai (C. natsudaidai), a susceptible cultivar. To develop markers, we focused on structural variation (copy number variation, CNV, and presence/absence variation, PAV). It has been well documented that CNV and PAV of defense-related genes are associated with resistant cultivars. Using a read depth approach following next-generation sequencing, we performed genome-wide analysis of CNV and PAV in two varieties. As a result, 633 genes showing at least two times difference between the mapping reads from two varieties and 61 genes showing presence of the mapping reads in only either one of them were screened. Visual inspection using the Integrative Genomics Viewer (IGV) was performed and experimental validation is being investigated. Interestingly, one of PAV candidates showed polymorphism in ‘Kiyomi’ and ‘Natsudaidai’ as well as other resistant and susceptible cultivars. Our results suggest a necessity for the detection of structural variation and indicate that the candidates may be useful for molecular breeding for citrus canker resistance and understanding disease resistance mechanism.

In this study, the concentrations of isoflavones, anthocyanins and total phenol content (TPC) in 19 soybean mutant lines changed seed coat color from yellow to black or brown were determined. Among 19 soybean mutant lines, 5 soybean mutant lines with black pigment were detected 3 anthocyanins (delphinidin-3-O-β-D-glucoside, D3G; cyaniding 3-O-β -D-glucoside, C3G; petunidin 3-O-β-D-glucoside, Pt3G). The highest concentration of anthocyanins among 5 soybean mutant lines was D-16 (1280.0 ± 19.4 mg/100g seed coat) derived from cv. Danbaek. Although isoflavone contents of all soybean mutant lines showed lower levels compared to original cultivars, glycitein was detected only 5 soybean mutant lines (DP-37-2, DP-38, DP-39, DP-40, and DP-41 derived from cv. Daepung). In TPC of 19 soybean mutant lines, DP-10 was increase levels compared to original cultivar, while DP-37-2, DP-40, and DP-41 were decrease levels of TPC. Using reduction of DPPH, we measured the free radical scavenging activity (FRSA) among 19 soybean mutant lines. Five black and 4 brown soybean mutants showed significant increase in FRSA. On the basis of these results, it was concluded that gamma irradiation may change the isoflavone, anthocyanin, and total phenol contents of soybean. These mutant lines using in this study can be useful for the breeding of soybean varieties altering the nutritional values.

Ramie (Boehmeria nivea L. Gaudich.) is a hardy perennial herbaceous plant of the Urticaceae family and has been grown as a fiber crop in several countries including Korea for many centuries. Ramie leaves also have been traditionally used as a major ingredient of a type of rice cake called ‘Song-pyun’ in the Southwest area of Korea, especially Yeong-Gwang province. Despite its economic importance, the molecular genetics of ramie have not been studied in detail yet. Genetic resources of ramie were widely collected from domestic local sites by Bioenergy Crop Research Center (RDA) and Yeong-Gwang Agricultural Technology Center. For the systematic and efficient management of the genetic resources, we developed microsatellite molecular markers of ramie. To do this, we generated microsatellite-enriched genomic DNA libraries using magnetic bead hybridization selection method. 216 contigs containing microsatellite repeat motif were generated using nucleotide sequences of 376 clones from the libraries. Primer sets were designed from the flanking sequences of the repeat motif. Finally, we selected 26 microsatellite markers, possibly showing polymorphism among the genetic resources. Results on the genotype analysis of the ramie genetic resources using the microsatellite markers will be presented.

A new graft cactus (Gymnocalycium marsoneri x mihanovichii x G. mihanovichii) cultivar, 'Hwangjo' was developed from a interspecific cross between an orange colored G. marsoneri x mihanovichii breeding line '9834036', a line with characteristics of vigorous growth and stable color under the strong sun light, and an orange colored graft cactus (G. mihanovichii) breeding line 'IG177' and through line selection at the National Horticultural Research Institute, Rural Development Administration in 2005. It was developed with the aim to breed a bright and vivid orange colored cultivar. Its characteristic evaluation was conducted three times during 2003 to 2005. The color of both body and tubercle was yellow. The shape of globe was flattened round and it had 8 to 10 deep ribs. The spine was semi erect, medium sized and grayish brown color. Growth was faster than the comparison cultivar, 'Hugwang'. Propagation capability was excellent, setting 15 tubercles per globe. Characteristics of the cultivar could be maintained by vegetative propagation. Strong sun light and virus infection should be avoided.

A new graft cactus (Gymnocalycium mihanovichii) cultivar, 'Simhong' was developed from a cross between graft cactus breeding line 'Keumhong', a cultivar revealing a red colored globe with yellow colored tubercles, and a dark red line '9502052' and consecutive line selection at the National Horticultural Research Institute, Rural Development Administration in 2005. It was developed with the aim to breed a dark red cultivar. Its characteristic evaluation was conducted three times during 2003 to 2005. The color of both body and tubercle was dark red. The shape of the globe was flattened round and it had 8 to 10 deep ribs. The spine was medium sized, straight and dark brown. Its growth was faster than the comparison cultivar, 'Jinhong'. Propagation capability was excellent, setting 12 tubercles per globe. Characteristics of the cultivar could be maintained by vegetative propagation. Strong sun light and virus infection should be avoided.

A new graft cactus (Gymnocalycium mihanovichii x G. marsoneri x mihanovichii) cultivar, 'Yeonsim' was developed from a interspecific cross between two black with pink graft cactus (Gymnocalycium mihanovichii) breeding line '9603026' and G. marsoneri x mihanovichii breeding line 'BP', a line with the characteristics of wide spacing between ribs, and vigorous growth and through line selection at the National Horticultural Research Institute, Rural Development Administration in 2005. It was developed with the aim of breeding cultivars with new colors and globe shapes. Its characteristic evaluation was conducted during 2003 to 2005. The color of both body and tubercle was pink. The shape of the globe was flattened round and it had 7 to 9 deep ribs. The spine was short, straight and brown color. Growth was faster than the comparison cultivar, 'Yeonhwa'. Propagation capability was excellent, setting 12 tubercles per globe. Characteristics of the cultivar could be maintained by vegetative propagation. Strong sun light and virus infection should be avoided.

A new graft cactus (Gymnocalycium mihanovichii) cultivar ‘Suyeon’ was developed from a cross between two pink with black breeding lines ‘9603010’ and ‘9603026’, by line selection at the National Horticultural Research Institute, Rural Development Administration in 2005. It was developed with the aim of breeding a cultivar showing vigorous growth and multiple colors. A line ‘9603010’ was derived from a cross between a black line and a pink line. Investigation into characteristics was conducted three times from 2003 to 2005. The color of both body and tubercle was pink with black. The shape of the globe was round and it had 7 to 9 deep ribs. The spine was short and straight, and its color was brown. Growth was retarded compared to a comparative cultivar ‘Ojack’, on the other hand, showed good propagation capability, setting 11 tubercles per globe. Characteristics of the cultivar could be maintained by vegetative propagation. Growers should take care to protect them from strong sun light and virus infection.

Molecular genetic markers have wide applicability for a various genetic analyses, and genetic mapping with PCR-based markers has identified many loci in the rice genome. This study was conducted to develop a genetic map of rice based on SSR and MITE-AFLP

A new soybean variety “Geomjeongkong 3” was released by the National Crop Experiment Station, RDA, in 2001. This variety was derived from the cross between Shinano-kuro and SNU78010, the elite breeding line with large seed, in the summer season, 1990. The

Genetic diversity and phylogenetic relationships among five cultivated Capsicum species were assessed using both mor-phological characters and DNA finger-printing patterns. Based on the clustering analysis using morphological characters withineach species