본 연구에서는 좁은잎천선과 잎 추출물 및 극성별 용매 분획물의 항산화 활성을 검색하고 유효성분을 분리하여 화학구조를 규명하였다. 좁은잎천선과 잎 추출물 및 용매 분획물의 DPPH 및 ABTS+ 라디칼 소거 활 성을 측정한 결과 에틸아세테이트 분획물에서 우수한 라디칼 소거 활성을 확인하였다. 활성이 좋은 에틸아세테 이트 분획물에서 유효성분을 찾고자 vacuum liquid chromatography (VLC), silica gel column chromatography를 실시하였으며, 분리된 화합물은 1H 및 13C NMR 데이터 분석 및 문헌치 비교를 통하여 총 5개의 화합 물을 동정하였다. 분리된 화합물은 monoolein (1), oleic acid (2), lutein (3), afzelechin (4), catechin (5)으 로 확인되었으며 이들은 모두 좁은잎천선과에서 처음으로 분리된 화합물이다. 분리된 화합물에 대한 DPPH 및 ABTS+ 라디칼 소거 활성 실험 결과 afzelechin (4) 및 catechin (5)에서 활성이 우수하게 나타났으며 특히 catechin (5)의 경우 대조군인 비타민 C보다 더 좋은 라디칼 소거 활성이 있음을 확인하였다. 또한 HPLC 분석 을 통해 좁은잎천선과 잎에서 분리된 catechin의 함량을 확인한 결과 추출물에서 3.8 mg/g, 에틸아세테이트 분획물에서 20.8 mg/g이 함유되어 있는 것으로 확인되었다. 이상의 연구 결과로부터 좁은잎천선과 잎을 이용한 천연 항산화 소재로의 개발이 가능할 것이라 사료된다

본 연구에서는 세토카 가지 정유의 주요 성분을 분석하고 이들의 항균, 항염 및 세포독성 실험을 진행하 였다. 세토카는 제주도에서 널리 재배되고 있는 감귤류의 품종이다. 세토카 나무의 가지는 간벌 작업으로 인해 대부분 폐기되고 있고, 이러한 폐자원의 활용은 최근 많은 관심을 받고 있다. 정유 성분은 세토카 가지의 에탄올 추출물을 호호바오일로 처리하여 얻었다. 세토카 정유의 주요 성분은 ethyl linoleate (64.1%), ethyl palmitate (16.5%), neophytadiene (11.1%) 및 β-citronellol (5.1%)임을 확인하였다. 이들의 항균활성을 확인 하기 위하여 피부 관련 미생물에 대한 paper disc 확산법을 실시한 결과 Staphylococcus aureus 및 Propionibacterium acnes에서 좋은 항균활성을 보였다. 또한 항염활성을 확인하기 위해 lipopolysaccharide (LPS)로 염증이 유도된 대식세포에서 nitric oxide (NO) 생성량을 측정한 결과 세토카 정유 성분은 농도의존적 으로 NO 생성을 저해하였다. WST-1 분석법을 이용하여 세포독성을 측정한 결과 RAW 264.7 macrophage 및 HaCaT 각질형성세포에서 세포생존율이 무처리 대조군과 비슷한 결과를 보였다. 이러한 결과를 바탕으로 세 토카 가지 정유는 세포독성이 없으면서 염증억제 및 항균효과가 있음을 확인하였으며, 이를 응용한 화장품소재 로써의 개발이 가능할 것으로 여겨진다.

Early life stage mortality in fish is one of the problems faced by loach aquaculture. However, our understanding of immune system in early life stage fish is still incomplete, and the information available is restricted to a few fish species. In the present work, we investigated the expression of immune-related transcripts in loach during early development. In fishes, recombination-activating gene 1 (RAG-1) and sacsin (SACS) have been considered as immunological function. In this study, the expression of the both genes was assessed throughout the early developmental stages of loach using real-time PCR method. maRAG-1 mRNA was first detected in 0 dph, observed the increased mostly until 40 dph. Significant expression of maRAG-1 was detected in 0 to 40 dph. These patterns of expression may suggest that the loach start to develop its function after hatching. On the other hand, maSACS was detected in unfertilized oocyte to molura stages and 0 to 40 dph. maSACS mRNA transcripts were detected in unfertilized oocytes, suggesting that they are maternally transferred.

Ionizing radiation directly and indirectly affects gene expression within the plant genome. To access the physiological response of rice to different types of ionizing radiation, rice seeds were exposed to gamma-ray and ion beam radiation. Exposure to ionizing radiation dramatically decreased the shoot length compared with non-irradiated plants. Fluorescence-activated-cell-sorting (FACs) was used to measure DNA contents. There were significant correlations of dose-dependent between irradiated plant and non-irradiated plant. The radicals induced by the ionizing radiation in the plant could be observed by electron spin resonance (ESR). It was confirmed that the number of free radicals in cell was greatly increased all irradiated plants than non-irradiated plant. A significant positive correlation was shown between ionizing radiation dose and signal intensity. In order to determine the Genetic diversity, AFLP analysis was conducted with the irradiated plant and non-irradiated plant. Based on band patterns, the cluster analysis was conducted to evaluate the genetic variation by using the UPGMA (Unweighted Pair Grouping Method of Averages). Genetic diversity of irradiated plants by low dose ion beam was the closest non-irradiated plant and irradiated by high dose gamma-ray was the furthest from non-irradiated. We describe the detailed methods of ionizing irradiation and discuss its applications in genetic research as well as plant breeding.

Ionizing radiation affects gene expression from plant genomes. To monitor the genome-wide transcriptional changes induced by three types of ionizing radiation, we used the rice RNA sequencing to identify genes that are up- or down-regulated by gamma rays (GAs), proton (PRs) and ion beams (IBs). The Oryza sativa jacalin-like lectin domain containing proteins (OsJAC1) gene was highly induced by GAs, PRs and IBs. OsJAC1 was selected based on the expression patterns of a genome-wide dataset of RNA sequencing. Many jacalin-related lectin genes have been shown to be associated with disease resistance, biotic and abiotic stress signaling. Therefore, we studied its expression pattern in response to different abiotic stress and phytohormone treatments. The expression patterns of OsJAC1 under two different abiotic stress conditions (salt and heat stress) and phytohormones (salicylic acid and methyl jasmonate) were examined. The transcripts of OsJAC1 were significantly induced in response to abiotic stress conditions, including salt and heat treatments. In addition, it was induced in response to the salicylic acid and methyl jasmonate treatments, respectively. To investigate the sub-cellular localization of OsJAC1, the gene was expressed as a fusion protein tagged with GFP, in tobacco leaf epidermis and examined under confocal microscope. The OsJAC1 was clearly localized at the nucleus. These results provide critical insights into the molecular functions of the rice jacalin-like lectin domain containing proteins as receptors of external signals.

‘Tocomi-1’, a new japonica rice cultivar derived from a 200 Gy gamma ray irradiation with high tocopherol content and red pericarp. The local adaptability test of MRXII-1001-1 was carried out from 2012 to 2014 and it was named as ‘Tocomi-1’ in 2014. This variety is medium matured with heading date of August 12 in honam plain area of Korea. This variety is about 80 cm tall culm length and 106 spikelets per panicle. Its 1,000 grain-weight of rice seeds is 25.4 g. The yield potential of this variety is about 5.15 MT/ha in local adaptability test for three years. This variety exhibited greater seed longevity than the Donganbyeo, indicating a crucial role for tocopherols in maintaining viability during quiescence, and displayed faster seedling growth during the early growth stage. Tocopherol contents was 50% higher than the Donganbyeo. To study the molecular mechanism underlying vitamin E biosynthesis, we examined the expression patterns of seven rice genes encoding vitamin E biosynthetic enzymes. Accumulation levels of the OsVTE2 transcript and OsVTE2 protein in the ‘Tocomi-1’ were significantly higher than in the Donganbyeo. Sequence analysis revealed that the ‘Tocomi-1’ harbored a point mutation in the OsVTE2 promoter region, which resulted in the generation of MYB transcription factor—binding cis-element. These results help identify the promoter regions that regulate OsVTE2 transcription, and offer insights into the regulation of tocopherol content in ‘Tocomi-1’.

To successful molecular breeding, identification and functional characterization of breeding related genes and development of molecular breeding techniques using DNA markers are essential. Although the development of a useful marker is difficult in the aspect of time, cost and effort, many markers are being developed to be used in molecular breeding and developed markers have been used in many fields. Single nucleotide polymorphisms (SNPs) markers were widely used for genomic research and breeding, but has hardly been validated for screening functional genes in olive flounder. We identified single nucleotide polymorphisms (SNPs) from expressed sequence tag (EST) database in olive flounder; out of a total 4,327 ESTs, 693 contigs and 514 SNPs were detected in total EST, and these substitutions include 297 transitions and 217 transversions. As a result, 144 SNP markers were developed on the basis of 514 SNP to selection of useful gene region, and then applied to each of eight wild and culture olive flounder (total 16 samples). In our experimental result, only 32 markers had detected polymorphism in sample, also identified 21 transitions and 11 transversions, whereas indel was not detected in polymorphic SNPs. Heterozygosity of wild and cultured olive flounder using the 32 SNP markers is 0.34 and 0.29, respectively. In conclusion, we identified SNP and polymorphism in olive flounder using newly designed marker, it supports that developed markers are suitable for SNP detection and diversity analysis in olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.

Molecular markers are useful for selecting to include superior character genetic like as strong immune system and rapid growth in fish. The marker is also very important part of breeding technology in Olive flounder (Paralichthys olivaceus). Single nucleotide polymorphisms (SNPs) marker is already in use widely for genomic research and breeding. But this SNPs marker hardly has been validated for screening functional genes in Olive flounder. We study identify single nucleotide polymorphisms (SNPs) on Expressed sequence tag (EST) database, develop usable SNP marker and apply to wild sample and cultured of olive flounder. As a result, Out of total 4.327 ESTs, 693contigs and 514 SNP from total contigs were detected while these substitutions include 297 transitions and 217 transversions. 144 developed markers were applied in 16 samples (wild 8, culture 8), Out of total marker, only 32 markers had detected polymorphic in sample. Polymorphism of 32 markers was observed in the variety genes region involved in immunity and protein synthesis. And the 32 marker were identified 21 transitions, 11 transversions, and indel was not detected in polymorphic SNPs. The analysis on heterozygosity by sample showed 0.34 in wild sample and 0.29 in cultured sample. In conclusion, we was identified SNP and Polymorphism by designed new marker, it supports that development marker is suitable for SNP detection and diversity analysis in Olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.

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.

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.

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

FGF9/16/20 signaling pathway specify the developmental fates of notochord, mesenchyme, and neural cells in ascidian embryos. Although a conserved Ras/MEK/Erk/Ets pathway is known to be involved in this signaling, the detailed mechanisms of regulation of FGF signaling pathway have remained largely elusive. In this study, we have isolated Hr-Erf, an ascidian orthologue of vertebrate Erf, to elucidate interactions of transcription factors involved in FGF signaling of the ascidian embryo. The Hr-Erf cDNA encompassed 3110 nucleotides including sequence encoded a predicted polypeptide of 760 amino acids. The polypeptide had the Ets DNA-binding domain in its N-terminal region. In adult animals, Hr-Erf mRNA was predominantly detected in muscle, and at lower levels in ganglion, gills, gonad, hepatopancreas, and stomach by quantitative real-time PCR (QPCR) method. During embryogenesis, Hr-Erf mRNA was detected from eggs to early developmental stage embryos, whereas the transcript levels were decreased after neurula stage. Similar to the QPCR results, maternal transcripts of Hr-Erf was detected in the fertilized eggs by whole-mount in situ hybridization. Maternal mRNA of Hr-Erf was gradually lost from the neurula stage. Zygotic expression of Hr-Erf started in most blastomeres at the 8-cell stage. At gastrula stage, Hr-Erf was specifically expressed in the precursor cells of brain and mesenchyme. When MEK inhibitor was treated, embryos resulted in loss of Hr-Erf expression in mesenchyme cells, and in excess of Hr-Erf in a-line neural cells. These results suggest that zygotic Hr-Erf products are involved in specification of mesenchyme and neural cells.

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.

Manganese () is a trace element that is essential for normal physiology, and is predominantly obtained from food. Several lines of evidence, however, demonstrated that overexposure to exerts serious neurotoxicity, immunotoxicity and developmental toxicity, particularly in male. The present study aimed to evaluate the effect of 0, 1.0, 3.3, and 10 mg/kg/day doses of on the reproductive organs in the immature female rats. Rats (PND 22; S.D. strain) were exposed to () dissolved in drinking water for 2 weeks. The animals were sacrificed on PND 35, then the tissues were immediately removed and weighed. Histological studies were performed using the uteri tissue samples. Serum LH and FSH levels were measured with the specific ELISA kits. Body weights of the experimental group animals were not significantly different from those of control group animals. However, ovarian tissue weights in 1 mg and 3.3 mg dose groups were significantly lower than those of control animals (p<0.05 and p<0.01, respectively). Uterine tissue weights of 3.3 mg dose groups were significantly lower than those of control animals (p<0.01), while the 1 mg dose and 10 mg dose failed to induce any change in uterine weight. Similarly, only 3.3 mg dose could induce the significant decrease in the oviduct weight compared to the control group (p<0.05). Non-reproductive tissues such as adrenal and kidney failed to respond to all doses of exposure. The uterine histology revealed that the exposure could affect the myometrial cell proliferation particularly in 3.3 mg dose and 10mg dose group. Serum FSH levels were significantly decreased in 1mg dose and 10 mg groups (p<0.05 and p<0.01, respectively). In contrast, treatment with 1 mg dose induced a significant increment of serum LH level (p<0.05). The present study demonstrated that exposure is capable of inducing abnormal development of reproductive tissues, at least to some extent, and altered gonadotropin secretions in immature female rats. Combined with the well-defined actions of this metal on GnRH and prolactin secretion, one can suggest the might be a potential environmental mediator which is involved in the female pubertal process.

Molecular genetic markers have been widely used as powerful tools for analyzing the genome. In particular, SSR markers have practical applications in breeding systems because they can be used in high-throughput analyses for genetic mapping, heritable diversity testing, purity analysis, and marker-assisted selection. Currently, due to technical advances in DNA sequencing, large sequence databases are available for large-scale SSR mining and marker development. Here, we describe an automated method, the SSR Finder program, for SSR discovery in the onion sequence database, and primer design for amplifying the detected SSRs. A total of 1,049 SSR primers were obtained for genetic purity testing, and 100 SSR sets were analyzed in 14 bulb onion breeding lines using clustering analysis. A total of 20 selected markers from screening of all 1,049 SSR primers, were finally applied for genetic purity testing in three breeding lines, NW1, NW9, and NW10. The initial tests showed that 15%, 71%, and 97% of individuals within NW1, NW9, and NW10, respectively, were genetically homogeneous. These markers produced using the SSR Finder will be useful for investigating the genetic purity of onion breeding lines.

Ionizing radiation is known to cause chromosomal alterations such as inversions and deletions and affects gene expression within the plant genome. To monitor the genome-wide transcriptome changes by ionizing radiation, we used rice Affimetrix GeneChip microarray to identify genes that are up- or down regulated by gamma-ray (200 Gy, 60Co source), cosmic-ray and ion beam (40 Gy, 220 MeV carbon ion). The overall expression patterns between gamma-ray and ion beam were similar but cosmic-ray was regulated differently. Combined results from all 3 radiations identified 27 up-regulated genes and 188 down regulated genes. These results mean the induction of similar mechanism changes in treatments of gamma ray and ion beam. However the different expression in treatment of cosmic-ray might be due to the other environmental conditions. Among the commonly up- or down- regulated genes, we chose highly up- or down- regulated several genes and confirmed its regulation in response to ionizing radiation exposure by RT-PCR analysis. Moreover, we showed that specific co-expression networks of candidate radio marker genes by ARACNE algorithm. Our results present profiles of gene expression related to different ionizing radiation and marker gene to predict sensitivity to ionizing radiation, such as GS (glutelin subunit) and FBX322.

이전 연구에서 저자들은 여뀌 추출물의 항산화, 항노화, 항균 활성 및 여뀌 추출물을 함유한 크림의 보습 효과에 대한 결과를 보고한 바 있다. 본 연구에서는 여뀌 추출물과 그 주요 성분인 isoquercitrin의 사람 적혈구와 HaCaT 세포에서의 세포 보호 효과를 측정하였다. 여뀌 추출물의 피부 전달시스템으로 에토좀 및 탄성 리포좀을 제조하여 입자크기,포집효율, 안정성 및 피부 흡수 증진 능력을 평가하였다. 적혈구 광용혈에서 여뀌 추출물과 isoquercitrin은 5 μg/mL의 농도에서 α-토코페롤보다도 큰 세포보호효과를 나타내었다. 여뀌 추출물은 HaCaT 세포에 대해 50 μg/mL의 농도에서 독성을 나타내지 않았다, UVB 400 mJ/cm2를 HaCaT cell에 조사하였을 때, 여뀌 추출물은 농도 의존적(12.5 ~ 50 μg/mL)으로 자외선으로부터 세포를 보호하였고, 25 μg/mL의 농도에서 양성 대조군(세포 생존율, 36 %)에 비하여 큰 세포보호 활성(생존율, 90 %)을 나타내었다. 0.04 % 여뀌 추출물을 담지한 에토좀의 입자 크기는 173.0 nm, 포집효율은 55.58 %이였다. 에토좀 제조 후 일주일동안 안정한 단분산 형태를 나타내었다. 피부 투과 실험 결과 에토좀은 일반 리포 좀이나 에탄올 용액에서보다도 더 큰 피부 투과능을 보여주었다. 0.1 % 여뀌 추출물을 담지한 탄성 리포좀의 최적의 제형은 인지질 대 계면활성제(Tego®care 450)의 비율이 95 : 5으로 확인되었다. 0.1 % 여뀌 추출물 함유한 최적의 탄성 리포좀의 입자크기는 176.5 nm, 가변형성은 16.4, 포집효율은 68.8 %이었다. 여뀌 추출물을 담지한 탄성 리포좀은 계면 활성제가 포함되지 않은 제형보다 더 큰 피부 투과능을 나타내었다.