To define whole genome-level of structural variation by ionization energies and radiation doses in plant, the seeds of Ilpum rice cultivar were acutely irradiated with gamma rays (100Gy, 200Gy, and 400Gy) and ion-beams (20Gy, 40Gy, and 80Gy), respectively. Six M1 rice plants were re-sequenced by Hi-Seq2500 with Ilpum cultivar as control. The average sequencing coverage of the individuals was 10.6X, and the average mapping rate to the rice reference genome (IRGSP-1.0) sequence was 96.95%. The individual plants were irradiated with gamma-400Gy and ion-50Gy had highest variation of SNP with 471,837 and 469,147, respectively. The number of insertion/deletion was 77,500 and 77,106, the synonymous and frame-shift were 7,859 and 7,763 in above two individuals. Although high genome variation shown between Ilpum cultivar and irradiated individuals, there were non-correlation between number of variation and radiation doses. However, five individuals, except ion-20Gy, showed 33 common variant blocks (CVBs) spanning 6 Mb in whole rice genome (1.6%). The CVBs were distributed on 12 rice chromosomes, Chromosome 6 had biggest CVB (5 blocks, 1.3Mb), whereas chromosome 9 had smallest CVB (0.01Mb). Total five hundred fifty one genes were in CVBs which can regard radiation sensitive genes or may be regarded as radiation hot spots in rice genome. This study will contribute to the improvement of the radiation mutation breeding research in genetic and genomic aspect.

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.

꽃게 조미소재, 야채, 천연조미료 등을 활용하면서, 편의 성을 부여하기 위하여 꽃게풍미 볶음밥을 제조하였다. 꽃 게풍미 볶음밥의 저장 중 품질변화를 조사하기 위해 –20, 4, 25℃에서 6주 동안 저장하면서 pH, 산도, 총균수, TBA, VBN, 관능적 기호도를 조사하였으며, 총균수 결과를 활용 하여 볶음밥의 유통 기한을 예측하였다. pH 변화를 조사한 결과, -20℃에 저장된 볶음밥의 경우 저장 0주째 pH는 7.74 이었고 저장 6주에는 pH 7.76으로 나타나 저장 중 변화가 거의 없는 것으로 나타났다. 적정산도를 조사한 결과, -20℃ 에 저장된 볶음밥의 경우 저장 0주째에 0.4%이었고 저장 6주에는 0.4%로 거의 변화가 없었다. 반면, 4℃ 와 25℃ 의 저장 6주에는 각각 2.7%, 4.6%의 수준으로 나타났다. TBA 값을 측정한 결과, -20℃에 저장한 볶음밥의 경우 저장 0주째 1.07 μg/g이었고, 저장 6주까지 1.39 μg/g으로 나타나 저장 중 다소 증가하는 경향을 보였다. 휘발성염기질소를 조사한 결과, -20℃에 저장된 볶음밥의 경우 저장 0주째에 26.60 mg%이었고, 저장 6주에는 28.14 mg%로 나타나 신선 도가 유지되는 것으로 사료되었다. 4℃와 25℃의 저장 6주 에는 각각 65.15 mg%, 68.00 mg% 수준으로 나타났다. 총균 수의 변화를 조사한 결과, -20℃ 저장된 볶음밥의 경우 저장 0주째 3.58 log CFU/g이었고 저장 6주째에는 4.11 log CFU/g으로 나타나 미생물변화는 안정적인 것으로 관찰되 었다. 관능적 기호도를 조사한 결과, -20℃에 저장된 볶음밥 의 경우 종합적 기호도의 변화를 보았을 때, 저장초기 7.8이 었고 6주째에는 6.7으로 나타나 저장 기간 동안 처리구 중 가장 높은 평가를 받았다. 25℃의 경우 저장초기 7.5이었으 나, 저장 6주째 1.0 수준의 평가를 받아 저장온도 및 저장기 간 동안 가장 낮은 평가를 받았다. 미생물 결과를 토대로 볶음밥의 유통기한을 예측한 결과, -20℃에 저장할 경우 14.9주, 4℃에서 저장할 경우 0.4주 인 것으로 각각 조사되었 다. 이상의 결과를 종합해 볼 때, 꽃게풍미 볶음밥은 -20℃에 서 저장하는 것이 미생물학적 및 관능적인 측면에서 저장성 이 좋을 것으로 사료되었고, 편의식품화로서의 이용 가능 성이 있을 것으로 생각되었다.

Gamma irradiation has been used as a tool for plant mutation breeding to select new cultivar with improved characteristics. Generally, the irradiation of seeds with high doses of gamma rays disturbs the synthesis of protein, hormone balance, and enzyme activity. And also, high dose of gamma rays to reduce plant height, number of tiller, and root length, although the effect of gamma-irradiated plants may depend on the species and cultivar or stress conditions. Biological effects of radiations can be divided into two types according to dose range and periods of exposure. Acute irradiation represents exposure to high-dose of irradiation over short period time, whereas the chronic irradiation is comprised of exposure to low doses of radiation over extended period of time. To compare the effects of acute and chronic exposure to ionizing radiation on two wheat cultivars (K4191 and Geumgangmil), we measured their germination rate, seedling height, and root length. In order to understand the influence of antioxidant-related genes and DNA repair-related genes, we used qRT-PCR methods to identify their expression levels. To study the behavior of a radiation-induced free radical, gamma-irradiated seeds were used for ESR spectroscopy. Plant growth pattern was showed positive correlation with ESR results. This study indicates that low level chronic radiation exposure is even more serious effects than short doses of high level radiation according to different wheat cultivar.

Chronic gamma irradiation can be used an alternative mutation breeding methods for induction of many useful mutants. Seedlings of purple-colored wheat plants were irradiated with wide range doses of chronic gamma-rays (20, 25, 30, 40, 50, 70, 100, 125, 150, 200, 250, 300 Gy) during 6 weeks at gamma-phytotron in the Korea Atomic Energy Research Institute, respectively. To identify the biological responses purple-colored wheat, we examined the plant height, chlorophyll, carotenoid and total anthocyanin contents in leaf. Plant growth, chlorophyll and carotenoid contents in leaf were decreased when the dose rate increased. Anthocyanin contents were increased with the increase of the radiation dose until 50 Gy treatment. To confirm the real contents of anthocyanin, we also investigated cyanidin-3-glucoside in purple-colored wheat leaf by using UPLC analysis. These results indicate that anthocyanin accumuation was observed under chronic gamma irradiation.

Miraculin can modify a sour taste into a sweet taste and is an alternative to traditional sweeteners. Mirculin gene was introduced into the Miyagawa Wase Satsuma mandarin (Citrus unshiu Marc.) callus by Agrobacterum-mediated transformation to produce citrus transgenic plant. Transgenic plant candidates were selected via hygromycin resistance. Transformation was confirmed by Polymerase Chain Reaction, Southern blotting, and Western blot analysis. Expression of this gene in transgenic citrus resulted in the accumulation of miraculin protein in the leaves. Multiple Shoots of transgenic citrus planets were micrografted onto trifoliate rootstocks in the sterile soil. Plants were established in the greenhouse 2 years after planting.

Somatic embryogenic calli were obtained from different native citrus species in Jeju island, South Korea. Undeveloped ovules were cultured on 5 different media, respectively; MSI (MS, modified, with the addition of malt extract 500 mg･L-1 and sucrose 50 g･L-1 and agar 2 g･L-1), MSII (MS, modified, with the addition of malt extract 500 mg･L-1 , kinetin 1 mg･L-1 and sucrose 50 g･L-1 and agar 2 g･L-1), MSIII (MS, modified, with the addition of malt extract 500 mg･L-1, 6-benzyladenine, 3 mg･L-1 and sucrose 50 g･L-1 and agar 2 g･L-1), EME-S (MT, modified, with the addition of malt extract 500 mg･L-1 and sucrose 50 g･L-1 and agar 2 g･L-1), 1/2 EME-S (half concentration of MT marcronutrients, half concentratrion of BH3 marcronutrients, malt extract 500 mg･L-1, glutamine 1.55 g･L-1 and sucrose 50 g･L-1 and agar 2 g･ L-1). Embryogenic calli were induced in the surface of undeveloped ovules in different manners, depending on citrus species and culture conditions. Somatic embryos developed into plantlets with a high frequency. Citrus embryogenic calli can be applied widely to somatic hybridization, genetic transformation, and in vitro germplasm conservation

Cinnamyl alcohol dehydrogenase (CAD) catalyzes cinnamyl aldehydes into cinnamyl alcohols, the final step in lignin biosynthesis. In this studied the purification, identification and characterization of a new cinnamyl alcohol dehydrogenase gene isolated from Citrus platymamma hort. Ex Tanaka. We expressed CAD potential gene in E.coli and then characterized its features in variety of specificity aldehydes substrates. The recombinant CAD protein was shown highest efficiently catalytic toward cinnamyl and coniferyl aldehydes and the shown lowest efficiently catalytic toward sinapyl aldehydes. We used a new improved analytical HPLC method in CAD enzymatic assay for fast and accurately measurement in various aldehydes substrates. In conclusion, our studies indicated the enzymatic activity of cDNA cloned CAD protein from Citrus platymamma hort. Ex Tanaka.

Perilla frutescens (L.) is an annual herbaceous and ornamental plant in the Lamiaceae family. Perilla frutescens (L.) Britt.cv.Chookyoupjaso were irradiated using a 200 Gy gamma ray in 1995. By HPLC analysis, this new cultivar significantly induced isoegomaketone content compared with ‘Chookyoupjaso’ control. The phenotypical difference was the changed leaf color of the ‘Atom-Ketone’ from violet to green. The yield potential of this cultivar (106 kg/10a) was 1.83 folds higher than that of ‘Chookyoupjaso’ (57.65 kg/10a). The methanol extracts of ‘Atom-Ketone’ inhibit nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells. This extract was further partitioned using ethyl acetate (EtOAc), butanol (BuOH), and water. The EtOAc fraction (EF-Atom-Ketone) was evaluated for antiinflammatory activities. These results indicated that the EF-Atom-Ketone reduced NO production by inhibiting inducible nitric oxide synthase (iNOS) expression. The EF-Atom-Ketone treatment also significantly diminished expression of MCP-1 and IL-6. Therefore, ‘Atom-Ketone’ reveals the potential therapeutic use of bioactive

Perilla frutescens (L.) is an edible plant, not only used as s food ingredient, but also in skin cream, soaps, and medicinal preparetions. ‘Atom-Ros’, a perilla (Perilla frutescenc (L.) Britt. cv. Chookyoupjaso was developed in 1995 by 200 Gy gamma irradiation-mutagenesis. This new cultivar has high rosmarinic acid content more than two fold compare with ‘Chookyoupjaso’ control. The observed phenotypical difference was changed leaf color of the ‘Atom-Ketone’ from violet to green. The yield potential of this cultivar (123.5 kg/10a) was 2.14 fold higher than that of ‘Chookyoupjaso’ (57.65 kg/10a). The methanol extracts of ‘Atom-Ros’ were tested for inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophase cells. Atom-Ros showed significant inhibition of NO production. This rosmarinic acid extracted from ‘Atom-Ros’ has a good potential to be developed as an antioxidant agent.

May-Thurner syndrome is caused by blockade of local venous flow due to local vascular intimal proliferation, caused by repeated pulsatile compression of the iliac or iliofemoral vein between the iliac artery and the lumbar spine. In this case, we confirmed May-Thurner syndrome using lower extremity computed tomographic angiography and venography. However, on venography, it was impossible to distinguish the left iliac vein from the collateral vein; a thrombus was also seen, although some of the thrombus was not seen clearly. These problems were overcome with use of intravascular ultrasound. We report on intravascular ultrasound guided treatment of May-Thurner syndrome.

MFG-E8 (Milk fat globule-epidermal growth factor VIII), also called lactadherin or BA46, SED1 is a glycoprotein found in milk and mammary epithelial cells, it is a major protein component associated with milk fat globule membrane. Previously, our study showed that expression of MFG-E8 is gradually increased with hepatic differentiation of human embryonic stem cells (hESCs). Therefore, we hypothesized that MFG-E8 would be an early cancer stem cell marker, which may predict cancer progression. Our results showed that MFG-E8 was expressed in various human cancer cell lines such as HepG2, Hep3B, and Huh7. Production and secretion of the MFG-E8 were also confirmed in the conditioned media of those three cell lines using enzyme-linked immunosorbent assay. Next, we analyzed the MFG-E8 expression in 11 clinical cases of cholangiocellular carcinoma (CC) and 33 cases of hepatocellular carcinoma (HCC) by immunohistochemistry and examined the potential correlation with β-catenin and AFP, which are known cancer markers. According to hitological criteria, the progression of HCC and CC was evaluated and classified into high, low, metastatic, and well-, moderate-, poor-differentiated, respectively. Statistical analysis indicated that incidence of both HCC and CC is significantly associated with male compared to female (P<0.05). Tumor size also has positive correlation with age (r2=08948). Our immunohistochemistry data showed that MFG-E8 was expressed both HCC and CC tissue. Interestingly, the MFG-E8 expression was significantly increased with cancer progression (P<0.05) in both cases. Additionally, b-cateninexpression was increased and its localization was changed from membrane to cytoplasm and nucleus with the degree of HCC. Likely b-catenin, AFP was also increased with the degree of HCC but it was not correlated with severalty of CC. Importantly, both AFP and b-catenin were highly co-localized with MFG-E8 in HCC. These results suggest that MFG-E8 may have important physiological roles and its expression in HCC and CC would be considered as an important prognostic factor.

Copy number variations (CNVs) are considered major sources of genetic variation, and CNVs may influence phenotypic variation and gene expression. To detect CNVs, rice seeds were exposed with 100～400 Gy of gamma-ray (GA, 60Co), cosmic-ray (CR) by Russia ISS, and 30 and 40 Gy of ion beam (IB, 220 Mev carbon ion). After the exposed rice seeds were cultured in 1/2 MS medium for 14 days, they were used for array-based Comparative genomic hybridization (CGH) analysis using Agilent’s RICE CGH array. As a results, the highest number of CNVs (Gain 808 and Loss 24,080) were detected in the CR treatment, whereas GA100 (100 Gy of GA) was identified the least CNVs. Compared individual chromosome, the chromosome 8 and 11 were identified the highest CNVs, the chromosome 3 had the least CNVs. Most of identified CNVs existed in the range of 10～500kb. In particular, the same CNV locations among different types of ionizing radiation were observed in chromosome 12, and these CNVs contained the commonly 5 amplified genes, containing retrotransposon protein, NADH-ubiquinone oxidoreductase chain 3, heavy metal transport/detoxification protein domain containing protein, and 2 unknown proteins. Other studies were reported that Ty1 (Long Terminal Repeat-retrotransposon family 1) transcription and retrotransposition were induced by different environmental stresses such as ionizing radiation, UV-light exposure, DNA damage and nutrient starvation in Saccharomyces cerevisiae. Our results also show that retrotransposon protein (LOC_Os12g34016) was specifically amplified by different types of ionizing radiation.

VitE (tocotrienols and tocopherols) are micronutrients with antioxidant properties synthesized by photosynthetic bacteria and plants that play important roles in animal and human nutrition. A new mutant line, T1001-1, was isolated from in vitro mutagenized population by ionizing radiation and shown to have increased VitE contents. The total VitE content was 26% increased in the T1001-1 mutant seeds compare with cv. Dongan (wild-type). In addition, we showed that the mutant confers retarded seedling growth during the early seedling growth stage in rice. To study the molecular mechanism of VitE biosynthesis, we used the rice microarray to identify genes that are upor down-regulated in T1001-1 mutant. In addition, we identified differentially regulated pathway using MapMan analysis, which provides deep insight into changes in transcript and metabolites. Our results enhanced the transcription of genes involved in starch and lipid metabolism in T1001-1 mutant. To identify the molecular mechanisms of the events involving transcription factors in tocopherol accumulation, we compared the expression patterns of transcription factors. The AP2-EREBP, WRKY, C2H2 transcription factor were up-regulated, whereas the MYB family was down-regulated in T1001-1 mutant. Our results demonstrate change of important transcript in high level of VitE accumulating rice mutant.

To determine the expression levels of genes related to the salt stress response in rice, gene expression profiles were investigated through microarray analysis using the rice mutant line Till-II-877. There were no significant changes in physiological response under salt stress of the mutant increased less than that in the WT. The intensity of gene expression was analyzed and compared between the wild type and mutant lines using a microarray. Among the most significantly affected pathways, α-linolenic acid metabolism and linoleic acid metabolism (in lipid metabolism), fructose and mannose metabolism and glycolysis-gluconeogenesis (in carbohydrate metabolism), cysteine and methionine metabolism (in amino acid metabolism), and carbon fixation (in the energy metabolism of photosynthetic organisms) showed changes in gene expression levels under salt stress. These results further our understanding of the effects of salt stress in rice and may aid in the development of salt-tolerant rice cultivars.

Soluble sugar content in soybean seed is an important quality attribute for soyfood and feed. Usually, soluble sugars comprise 6 to 17% of total dry wt. in mature soybean seeds. In this study, 414 soybean mutant lines induced by gamma-ray were screened by colormetric assay, FACE (Fluorophore-assisted carbohydrate electrophoresis), and GC-MS to identify the change of soluble sugar contents. Among 414 soybean mutant lines, 12 mutant lines derived from three different soybean cultivars (Hwanggum, Paldal, and Bangsa) showed higher level of soluble sugar content compared to their original cultivars. However, 5 mutant lines derived from soybean landrace KAS 636-15 showed lower level in the colormetric assay. In FACE, 17 soybean mutant lines selected by colormetric assay also showed different band intensity compared with their original cultivars. However, there were no different soluble sugar patterns between soybean original cultivars and mutant lines. Finally, the variations of soluble sugar content in 17 soybean mutant lines were confirmed by using GC-MS. These mutant lines will be used for genetic study to find mutations of genes related soluble sugar biosynthesis.

Compared to wide ranges of genetic variation of natural populations, very limited Miscanthus cultivar has been released. This study was the first report on the development of Miscanthus cultivar by means of radiation breeding. Seeds of M. sinensis were initially exposed to gamma rays of 250 Gy for 24 h, generated from a 60Co gamma-irradiator. The irradiated seeds were sown and then the highly tiller-producing mutants were selected for this study. Biomass-related parameters including tiller number, plant height, stem diameter, and leaf number were measured. Ploidy level and internal transcribed spacer (ITS) were investigated to characterize the mutants compared to wild type (WT) Miscanthus. Plant height and tiller number were negatively related, where multi-tillering mutants were relatively short after 4 month growth. However stem diameter and leaf number were greater in mutants. All the materials used in this study were diploid, implying that the mutants with greater tiller numbers and stem diameter were not likely related to polyploidization. Based on the sequence of ITS regions, the mutants demonstrated base changes from the gamma irradiation where G+C content (%) was decreased in the ITS1, but increased in ITS2 when compared to WT sequence. ITS2 region was more variable than in ITS1 in the mutants, which collectively allows identification of the mutants from WT. Those mutants having enhanced tillers and allelic variations might be used as breeding materials for enhanced biomass-producing Miscanthus cultivars.

Citrus is one of the major fruits produced in Korea. There are about 20 species mainly grown in Jeju Island, Korea. Four representative species, which are quite different in the shape of leaf and the taste of fruit, were selected and were used to profile the transcriptomes. These species are ‘Miyagawa Wase’ (C. unshiu Marcov.) satsuma mandarin, ‘Kiyomi’ (C. unshiu Marcov. × C. sinensis) mandarin hybrid, ‘Dangyuja’ (C. grandis) and ‘Natsudaidai’ (C. natsudaidai). Classification of the up-regulated and down-regulated genes using the Cluster of Orthologous Groups of proteins (COG) database reveals that the number of genes included in each group differed significantly among the four species. Several genes that showed significant differences in expression on the microarray were selected and their expression patterns were examined by reverse transcription- ploymerase chain reaction. Metabolic genes such as tyrosine decarboxylase and β-glucosidase ligase were found to be highly expressed in Miyagawa Wase, relative to other species. On the other hand, the expression level of mannose phosphate isomerase was lower in Miyagawa Wase. An efflux pump gene was found to be up-regulated in Kiyomi, whereas cinnamyl-alcohol dehydrogenase was down-regulated. β-carotene 15,15’-dioxygenase, which is involved in the vitamin metabolism, was up-regulated in Natsudaidai. Interspecific differentiations of gene expression are analyzed in terms of the metabolic pathways and their possible roles in citrus species.