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Studies of Molecular Breeding Technique Using Genome Information on Edible Mushrooms

  • Kong, Won-Sik;Woo, Sung-I;Jang, Kab-Yeul;Shin, Pyung-Gyun;Oh, Youn-Lee;Kim, Eun-sun;Oh, Min-Jee;Park, Young-Jin;Lee, Chang-Soo;Kim, Jong-Guk
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.53-53
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    • 2015
  • Agrobacterium tumefaciens-mediated transformation(ATMT) of Flammulina velutipes was used to produce a diverse number of transformants to discover the functions of gene that is vital for its variation color, spore pattern and cellulolytic activity. Futhermore, the transformant pool will be used as a good genetic resource for studying gene functions. Agrobacterium-mediated transformation was conducted in order to generate intentional mutants of F. velutipes strain KACC42777. Then Agrobacterium tumefaciens AGL-1 harboring pBGgHg was transformed into F. velutipes. This method is use to determine the functional gene of F. velutipes. Inverse PCR was used to insert T-DNA into the tagged chromosomal DNA segments and conducting sequence analysis of the F. velutipes. But this experiment had trouble in diverse morphological mutants because of dikaryotic nature of mushroom. It needed to make monokaryotic fruiting varients which introduced genes of compatible mating types. In this study, next generation sequencing data was generated from 28 strains of Flammulina velutipes with different phenotypes using Illumina Hiseq platform. Filtered short reads were initially aligned to the reference genome (KACC42780) to construct a SNP matrix. And then we built a phylogenetic tree based on the validated SNPs. The inferred tree represented that white- and brown- fruitbody forming strains were generally separated although three brown strains, 4103, 4028, and 4195, were grouped with white ones. This topological relationship was consistently reappeared even when we used randomly selected SNPs. Group I containing 4062, 4148, and 4195 strains and group II containing 4188, 4190, and 4194 strains formed early-divergent lineages with robust nodal supports, suggesting that they are independent groups from the members in main clades. To elucidate the distinction between white-fruitbody forming strains isolated from Korea and Japan, phylogenetic analysis was performed using their SNP data with group I members as outgroup. However, no significant genetic variation was noticed in this study. A total of 28 strains of Flammulina velutipes were analyzed to identify the genomic regions responsible for producing white-fruiting body. NGS data was yielded by using Illumina Hiseq platform. Short reads were filtered by quality score and read length were mapped on the reference genome (KACC42780). Between the white- and brown fruitbody forming strains. There is a high possibility that SNPs can be detected among the white strains as homozygous because white phenotype is recessive in F. velutipes. Thus, we constructed SNP matrix within 8 white strains. SNPs discovered between mono3 and mono19, the parental monokaryotic strains of 4210 strain (white), were excluded from the candidate. If the genotypes of SNPs detected between white and brown strains were identical with those in mono3 and mono19 strains, they were included in candidate as a priority. As a result, if more than 5 candidates SNPs were localized in single gene, we regarded as they are possibly related to the white color. In F. velutipes genome, chr01, chr04, chr07,chr11 regions were identified to be associated with white fruitbody forming. White and Brown Fruitbody strains can be used as an identification marker for F. veluipes. We can develop some molecular markers to identify colored strains and discriminate national white varieties against Japanese ones.

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Novel Species Candidates Belonging to the Phyla Bacteroidetes, Firmicutes, and Actinobacteria Isolated from the Halla Mountain Wetlands (제주도 고산 습지에서 분리한 Bacteroidetes, Firmicutes, Actinobacteria 문에 속하는 신종후보 세균)

  • Choi, Ah-Young;Choi, Jae-Hee;Kang, Ji-Young;Choe, Jeong-Uk;Lee, Sang-Hoon;Kim, Ha-Neul;Yi, Ha-Na;Shin, Young-Min;Jahng, Kwang-Yeop;Lee, Hyune-Hwan;Kim, Kyu-Joong;Joh, Ki-Seong;Chun, Jong-Sik;Kim, Seung-Bum;Cho, Jang-Cheon
    • Korean Journal of Environmental Biology
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    • v.29 no.3
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    • pp.126-137
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    • 2011
  • Although Sumunmulbangdui wetland at the Halla Mountain in Jeju Island, a kind of montane wetlands, has been considered to bear high biodiversity, no study has been reported on the bacterial diversity. In this study, soil and water samples were collected from the wetland in order to isolate novel bacterial species. Bacterial strains belonging to the phyla Bacteroidetes, Firmicutes, and Actinobacteria were isolated after spreading soil and water samples onto solid agar media. The 16S rRNA gene sequences of the strains assigned to the three phyla were compared to those of type strains of the species in the phyla. The strains that showed less than 98.7% 16S rRNA gene sequence similarity to the validly published species were considered to be novel species candidates. A total of 32 strains were regarded as novel species candidates in the phyla Bacteroidetes, Firmicutes, and Actinobacteria. Diversity of novel species candidates was very low; the candidates were confined to only few genera. In the Bacteroidetes, 13 novel candidate species were affiliated with the genera Mucilaginibacter, Sphingobacterium, Pedobacter, Flavobacterium, and Chryseobacterium. A total of 13 novel candidate species that assigned to the genera Paenibacillus Lysinibacillus, and Bacillus were identified in the phylum Firmicutes. Only two candidate species that belonged to the genera Mycobacterium and Nocardia were excavated in the Actinobacteria. Cultural, physiological, and chemotaxonomic characteristics have been determined for the novel species candidates, and the characteristics are described in this study.

Selection of (Ac/Ds) insertion mutant lines by abiotic stress and analysis of gene expression pattern of rice (Oryza sativar L.) (비생물학적 스트레스 관련 벼 Ac/Ds 삽입 변이체의 선발 및 유전자 발현 분석)

  • Jung, Yu-Jin;Park, Seul-Ah;Ahn, Byung-Ohg;Yun, Doh-Won;Ji, Hyeon-So;Lee, Gang-Sup;Park, Young-Whan;Suh, Seok-Cheol;Baek, Hyung-Jin;Lee, Myung-Chul
    • Journal of Plant Biotechnology
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    • v.35 no.4
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    • pp.307-316
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    • 2008
  • Transposon-mediated insertional mutagenesis is one of powerful strategy for assessing functions of genes in higher plants. In this report, we have selected highly susceptible and tolerance plant by screening about high salt (3% NaCl) and cold stresses ($4^{\circ}C$) from F2 seeds of 30,000 Ac/Ds insertional mutagenesis lines in rice (Oryza sativa L. cv. Dongjin). In order to identify the gene tagging, insertion of Ds element was analyzed by Southern blot and these results revealed that 19 lines were matched genotype of selected lines with phenotype from the first selected 212 lines, and 13 lines have one copy of Ds elements. The Franking Sequence Tags (FSTs) of selected mutant lines showed high similarities with the following known function genes: signal transduction and regulation of gene expression (transpoter, protease family protein and apical meristem family protein), osmotic stress response (heat shock protein, O-methyltransferase, glyceraldehyde-3-phosphate dehydrogenase and drought stress induce protein), vesicle trafficking (SYP 5 family protein) and senescence associated protein. The expression pattern of 19 genes were analyzed using RT-PCR under the abiotic stresses of 9 class; 250mM NaCl, osmotic, drought, 3% $H_2O_2$, $100{\mu}M$ ABA, $100{\mu}M$ IAA, 0.1 ppm 2,4-D, $4^{\circ}C$ cold and $38^{\circ}C$ high temperature. Isolated knock-out genes showed the positive response about 250 mM NaCl, drought, $H_2O_2$, PEG, IAA, 2,4-D, ABA treatment and low ($4^{\circ}C$) and high temperature ($38^{\circ}C$). The results from this study indicate that function of selected knock-out genes could be useful in improving of tolerance to abiotic stresses as an important transcriptional activators in rice.

Isolation of Nontuberculous Mycobacteria by DNA Probe and Clinical Characteristics of Patients with NTM Pulmonary Disease (DNA probe를 이용한 비결핵항산균의 분리 및 폐질환자들의 임상적 특징)

  • Kim, Hee Kyoo;Kim, Yu Ri;Park, Jung Pil;Kim, Nang Hee;Ok, Chul Ho;Jung, Maan Hong;Jang, Tae Won;Jeong, Seok Hoon;Kim, Cheol Min;Park, Hee Kyung
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.3
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    • pp.248-256
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    • 2005
  • Background : Nontuberculous mycobacterial (NTM) infections are increasingly being recognized as a cause of chronic pulmonary disease. This study describes the prevalence of NTM species from clinical specimens and the clinical characteristics of NTM pulmonary disease. Material and Methods : The NTM isolated from March 2003 to December 2003 at the Kosin Medical Center were identified using an oligonucleotide chip containing the internal transcribed space (ITS) sequence. The medical records of the patients with the NTM isolates, who fulfilled the 1997 ATS diagnostic criteria for NTM pulmonary disease, were analyzed, retrospectively. Results : Twenty four species (24.2%) of NTM were isolated from 99 cultured AFB specimens. M. avium complex (MAC) (13 isolates), M. szulgai (3), M. kansasii (2), M. malmoense (2), M. abscessus (1), M. chelonae (1), M. scrofulaceum (1), and unclassified (1). Of the 23 patients with isolated NTM, 11 patients were found to be compatible with a NTM pulmonary infection according to the ATS criteria; MAC was found in 6 cases (54.5%), M. szulgai in 2 cases (18.2%), and M. abscessus, M. szulgai, M. kansasii and M. malmoense in 1 case each (9.1%). Ten patients (91%) were male and the median age at diagnosis was 61 years. In the pre-existing diseases, malignant disease was found in 6 cases including 5 patients with lung cancer, and history of old pulmonary tuberculosis was identified in 4 cases. The radiological patterns showed lung destruction lung in 3 cases, a cavitary mass in 3 cases, a nodular pattern in 2 cases, and reticulonodular, consolidation and a bronchiectasis pattern were in 1 case each. Conclusion : Various types of NTM pulmonary diseases were found in a tertiary hospital at Busan, Korea. The NTM pulmonary diseases were caused by MAC, M. szugai, M. kansasii, M. malmoense, M. abscessus, M. chelonae, and M. scrofulaceum in the order of frequency.

Phytosociological Changes of Pinus densiflora Forest Induced by Insect Damage in Kyonggi-do Area (경기도지방(京畿道地方) 적송림(赤松林)의 식물사회학적(植物社會學的) 연구(硏究))

  • Yim, Kyong Bin;Park, In Hyeop;Lee, Kyong Jae
    • Journal of Korean Society of Forest Science
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    • v.50 no.1
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    • pp.56-71
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    • 1980
  • To study the changes of vegetational composition of each plant successional stage of Pinus densiflora forest caused by pine gall midge damage, Thecodiplosis japonensis, located in Kyonggi-do area, the middle part of Korea, four districts, Anseong, Yongin, Gwangju, and Gapyeong, were selected according to the length of insect damage duration. The forest in the Anseong district was the noninfested one selected as the check. And forests in Yongin, Gwangju, and Gapyeong were the recent]y damaged, severely damaged, and damage prolonged ones in sequence. It was considered that this order could be in concord with the gradual transition of plant successional stage. In view of temperature, precipitation and soil factors, these four districts were sited within the same environment. Four survey plots were selected from each district and the distance between them was 3~4 km. Then again four subplots were set within a plot. Through needle accumulation after insect infestation, light intensity, soil moisture content, and the amount of soil organic matter were increased and these conditions could create a favorable environment enabling the new species invasion and their growth. These were naturally closely related with the increase in quantity of Pinus densiflora defoliation. After considerable time lapse, the quantities of above mentioned factors decreased with the development of the new substitute tree species. It was found that Quercus mongolica was an excellently substitute tree species come first in this studied area. Species diversity increased to a degree at the early stage of the duration of pine gall midge damage and decreased thereafter. As the same manner, similarity index value between plots in the same district decreased to a degree and increased thereafter. This was linked admittedly with the increase and decrease of tree species and evenness. Synthesizing above mentioned results, the process of plant succession studied quantitatively by changes of relative density and relative importance value, it can be concluded that Pinus densiflora forest gives way to Genus Quercus forest in which Genus Lespedeza and Genus Rhododendron species become as subdominance species after removal of the Pinus densiflora trees damaged by pine gall midge.

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${T_2}weighted$- Half courier Echo Planar Imaging

  • 김치영;김휴정;안창범
    • Investigative Magnetic Resonance Imaging
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    • v.5 no.1
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    • pp.57-65
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    • 2001
  • Purpose : $T_2$-weighted half courier Echo Planar Imaging (T2HEPI) method is proposed to reduce measurement time of existing EPI by a factor of 2. In addition, high $T_2$ contrast is obtained for clinical applications. High resolution single-shot EPI images with $T_2$ contrast are obtained with $128{\times}128$ matrix size by the proposed method. Materials and methods : In order to reduce measurement time in EPI, half courier space is measured, and rest of half courier data is obtained by conjugate symmetric filling. Thus high resolution single shot EPI image with $128{\times}128$ matrix size is obtained with 64 echoes. By the arrangement of phase encoding gradients, high $T_2$ weighted images are obtained. The acquired data in k-space are shifted if there exists residual gradient field due to eddy current along phase encoding gradient, which results in a serious problem in the reconstructed image. The residual field is estimated by the correlation coefficient between the echo signal for dc and the corresponding reference data acquired during the pre-scan. Once the residual gradient field is properly estimated, it can be removed by the adjustment of initial phase encoding gradient field between $70^{\circ}$ and $180^{\circ}$ rf pulses. Results : The suggested T2EPl is implemented in a 1.0 Tela whole body MRI system. Experiments are done with the effective echo times of 72ms and 96ms with single shot acquisitions. High resolution($128{\times}128$) volunteer head images with high $T_2$ contrast are obtained in a single scan by the proposed method. Conclusion : Using the half courier technique, higher resolution EPI images are obtained with matrix size of $128{\times}128$ in a single scan. Furthermore $T_2$ contrast is controlled by the effective echo time. Since the suggested method can be implemented by software alone (pulse sequence and corresponding tuning and reconstruction algorithms) without addition of special hardware, it can be widely used in existing MRI systems.

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Investigation of the Relationship between Interleukin-4 Promoter Polymorphism and Severity of Patients with Bronchial Asthma (천식 환자에서 증상의 정도에 따른 IL-4 유전자 다형에 관한 연구)

  • Kang, Sea-Yong;Shim, Jae-Jeong;Cho, Jae-Yun;Kwon, Young-Hwan;Lee, Seung-Yong;Kim, Je-Hyeong;Lee, Sang-Youb;Lee, So-Ra;Han, Seon-Ae;Kim, Han-Gyum;Kang, Kyung-Ho;Yoo, Se-Hwa;In, Kwang-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.3
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    • pp.529-535
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    • 1998
  • Background: Interleukin-4 plays an important role in pathogenesis of asthma, especially in developing atopy by means of switching B lymphocytes to produce IgE. It has been shown that there is polymorphism in the Interleukin-4 promoter region, transversion of cytosine to thymine at-598 from translation initiation site of IL-4 gene. There has also been quite a few works to reveal the role of the polymorphism of IL-4 gene in patients with asthma. We performed this investigation to determine the role of the polymorphism in the severity of symptoms of patients with asthma. We also examined the frequency and the type of the polymorphism in asthmatics compared with non-asthmatics as well. Method: The subjects enrolled in this study were 49 asthmatics and 33 non-asthmatics. All the asthmatics were classified as mild and moderate to severe by the NHLBI/WHO Workshop. DNA from both asthmatics and non-asthmatics was extracted, then performed ARMS(Amplification Refractory Mutation System) as well as RFLP using BsmFl restriction enzyme in order to confirm the polymorphism of Il-4 gene. Results: There was no significant difference in the occurrence of polymorphism of the IL-4 promoter sequence between asthm and non-asthma groups(P=0.7). Among those with polymorphisms, the number of C/C type was slightly more than C/T type in both asthmatics and non-asthmatics, 26 vs 21 in asthmatics and 18 vs 15 in non-asthmatics, which was, however, insignificant statistically. No significant relationship between the severity of asthma and the polymorphism was found(P=0.7). Conclusion: There was no significant difference between the severity of asthma and the IL-4 promoter polymorphism(P=0.709). Interestingly, the frequency of the polymorphism in both asthmatics as well as non-asthmatics was found to be even higher than that occurred in Caucasians. However, no significant difference in the frequency of the polymorphism was found in both groups.

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The Program Development of Nursing Activities for Community Health Promotion (지역사회 건강증진 간호활동의 프로그램 개발)

  • Park, Jeong-Hee
    • Research in Community and Public Health Nursing
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    • v.10 no.2
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    • pp.291-306
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    • 1999
  • Health promotion has come to the fore through new concept approach in consequence of the increase of chronic diseases. increase of medical cost and social trend of putting more emphasis on the individual responsibility for health. Studies of health promotion can be classified into two types: one is micro intervention method which is mainly focused on modifying individual life style and the other. macro intervention method in which they put another emphasis on the environment the individual is surrounded. in addition to modifying individual life style. This study belongs to the later. This study aims to develop nursing activities and program it for the purpose of community health promotion. The process of the study can be briefed as follows: to draw out nursing contents to intervene for community health promotion: to promote community health to develop nursing action indicators; to develop nursing action indicators for the development of main nursing activities. And those developed nursing activities are programmed systematically. The community health promotion program is composed of a hierarchical structure with nursing process that the nurses are supposed to apply to perform professional nursing. the level of nursing perform. the main items of nursing process. health promotion nursing indicators. health promotion nursing activities. The conclusion of this study according to objectives are as follows. First. community health promotion contents at individual and community level are remained revised and complemented and those at organizational level are developed. The developed main contents of community health promotion nursing are as follows. 1) Revised individual level nursing contents: 35 items. 2) Developed organizational level nursing contents: 24 items. 3) Revised community level nursing contents: 36 items. Second. for the development of the health promotion nursing action indicators. principles were set up and applied as follows. 1. Developed indicators should be provided with such qualities as comprehensiveness. diversity. developability, availability. practicability. 2. Developed indicators should be provided with functional abilities to measure the conditions and changes in any phenomena or state. inspect the development of the states. control the implementing program. evaluate the result of program and grasp what nurses should do. 3. Developed indicators should be provided with relevance and sequence. 4. Developed indicators should be undergo inspections from the expert. The developed community health promotion action indicators developed in this study. observing above mentioned principles. are total 330 indicators of 95 items. Third. when the main nursing activities were developed for each nursing action indicator. five priciples were set up in accordance with each nursing action indicators to decide main nursing activities. Main nursing activities developed observing those principles. are total 1273. Forth. for the programming of the developed nursing activities. three principles were set up. 1. The nursing activities are systematized in line with (nursing process) (nursing client) (key items of nursing process) (nursing action indicators for health promotion) (nursing activities). 2. The program is constructed in downward and hierarchical order. 3. The program is constructed not in relation to same level activities but in relation to high and low level activities. The process step of programming of developed main health promotion nursing activities are; Step 1. The Developed nursing action indicators are classified into nursing process. Step 2. The main nursing activities are allocated per each nursing action indicators. Step 3. The statement of main nursing activities are inspected. Step 4. The items of main nursing activities allocated by a certain nursing action indicators are sequenced. taking into consideration the elaborateness of activity. the sequency of activity. familiarity of activity. the difficulty of activity. the interest of activity. the frequency of activity. Step 5. The whole developed program should undergo comprehensive and critical inspections.

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Isolation and Characterization of Burkholderia cepacia EB215, an Endophytic Bacterium Showing a Potent Antifungal Activity Against Colletotrichum Species (탄저병균에 길항력이 우수한 식물내생세균 Burkholderia cepacia EB215의 분리 및 특성 규명)

  • Park Ji Hyun;Choi Gyung Ja;Lee Seon-Woo;Jang Kyoung Soo;Lim He Kyoung;Chung Young Ryun;Cho Kwang Yun;Kim Jin-Cheol
    • Microbiology and Biotechnology Letters
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    • v.33 no.1
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    • pp.16-23
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    • 2005
  • In order to develop a new microbial fungicide using endophytic bacteria for the control of anthracnoses occurring on various crops, a total of 260 bacterial strains were isolated from fresh tissues of 5 plant species. After they were cultured in broth medium, their antifungal activities were tested for in vivo antifungal activity against cucumber anthracnose caused by Colletotrichum orbiculare. As the results, liquid cultures of 28 strains showed potent antifungal activities more than $90\%$ against cucumber anthracnose. At 3-fold dilutions of liquid cultures, 18 strains inhibited the development of cucumber anthracnose of more than $70\%$. They were further tested for in vivo antifungal activity against red pepper anthracnose caused by C. coccodes and in vitro antifungal activity against C. acutatum, a fungal agent causing red pepper anthracnose. Among 18 strains, a bacterial strain EB215 isolated from cucumber roots displayed the most potent antifungal activity against Colletotrichum species. It was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, Biolog test and 16S rDNA gene sequence. It also controlled effectively the development of rice blast (Magnaporthe grisea), rice sheath blight (Corticium sasaki), tomato gray mold (Botrytis cinerea), and tomato late blight (Phytophthora infestans). Studies on the characterization of antifungal substances produced by B. cepacia EB215 are in progress.

An Analysis on Territorial Education of Geography Textbooks in Korea and Japan (한.일 지리교과서에 나타난 영토교육 내용 분석)

  • Lee, Ha-Na;Cho, Chul-Ki
    • Journal of the Korean association of regional geographers
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    • v.17 no.3
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    • pp.332-347
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    • 2011
  • This study is to analyze on territorial education described in geography textbooks in Korea and Japan. The following is the result that shows similarities and differences of the geography textbooks when it comes to territorial education. Korea and Japan have a contrasting territorial background. However, both countries start their territorial education by learning the location and shape of their country. Japanese geography textbooks focus on what people in the world think of Japan, but in case of Korea, the geography textbooks focus on how Koreans look at the world. In short, the territorial education in Japan try to emphasize Japan from the view point of the world. The next common ground is that the two countries provide territorial models in their geography textbooks in order to increase understanding. However, the Japanese students are provided with these territory models much earlier than Korean students and these models help them visualize and solidify their concept of territory. And, the two countries both put great importance on teaching territorial sea. In Japan, they try to include EEZ(Exclusive Economic Zone) in their territory. Considering these facts, it can be concluded that Japan is enlarging their concept of national territory as maritime territory. Lastly, after learning of territory the two countries both treat on territorial problems. But Korea treats passively territorial problem as such Dokdo, but Japan treats actively their territorial problems. Like that, the contents of territorial education described in geography textbooks in Korea and Japan are similar in terms of selection, but differ in quality in terms of organization. Therefore, future territorial education in Korea will be actively and successively done through succession and sequence of geography curriculum.

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