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Improving Corsican pine somatic embryo maturation: comparison of somatic and zygotic embryo morphology and germination

  • Wtpsk, Senarath;Shaw, D.S.;Lee, Kui-Jae;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.61-62
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    • 2003
  • Clonal propagation of high-value forest trees through somatic embryogenesis (SE) has the potential to rapidly capture the benefits of breeding or genetic engineering programs and to improve raw material uniformity and quality. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Several factors limit commercialization of SE for Corsican pine, including low initiation rates, low culture survival, culture decline causing low or no embryo production, and inability of somatic embryos to fully mature, resulting in low germination and reduced vigour of somatic seedlings. The objective was to develop a Corsican pine maturation medium that would produce cotyledonary embryos capable of germination. Treatments were arranged in a completely randomized design. Data were analyzed by analysis of variance, and significant differences between treatments determined by multiple range test at P=0.05. Corsican pine (Pinus nigra var. maritima) cultures were initiated on modified !P6 medium. Modifications of the same media were used for culture multiplication and maintenance. Embryogenic cultures were maintained on the same medium semi solidified with 2.5 g/l Gelrite. A maturation medium, capable of promoting the development of Corsican pine somatic embryos that can germinate, is a combination of iP6 modified salts, 2% maltose, 13% polyethylene glycol (PEG), 5 mg!l abscisic acid (ABA), and 2.5 g/l Gelrite. After initiation and once enough tissue developed they were grown in liquid medium. Embryogenic cell suspensions were established by adding 0.951.05 g of 10- to 14-day-old semisolid-grown embryogenic tissue to 9 ml of liquid maintenance media in a 250ml Erlenmeyer flask. Cultures were then incubated in the dark at 2022$^{\circ}$C and rotated at 120 rpm. After 2.53 months on maturation medium, somatic embryos were selected that exhibited normal embryo shape. Ten embryos were placed horizontally on 20 ml of either germination medium ($\frac{2}{1}$strength Murashige and Skoog (1962) salts with 2.5 g/l activated charcoal) or same medium with copper sulphate adjusted to 0.25 mg/1 to compensate for copper adsorption by activated carbon. 2% and 4% maltose was substituted by 7.5% and 13% PEG respectively to improve the yield of the embryos. Substitution of' maltose with PEG was clearly beneficial to embryo development. When 2% of the maltose was replaced with 7.5% PEG, many embryos developed to large bullet-shaped embryos. At latter stages of development most embryos callused and stopped development. A few short, barrel-shaped cotyledonary embryos formed that were covered by callus on the sides and base. When 4% of the maltose was removed and substituted with 13% PEG, the embryos developed further, emerging from the callus and increasing yield slightly. Microscopic examination of the cultures showed differing morphologies, varying from mostly single cells or clumps to well-formed somatic embryos that resembled early zygotic embryos only liquid cultures with organized early-stag. A procedure for converting and acclimating germinants to growth in soil and greenhouse conditions is also tested. Seedling conversion and growth were highly related to the quality of the germinant at the time of planting. Germinants with larger shoots, longer, straighter hypocotyls and longer roots performed best. When mature zygotic embryos germinate the root emerges, before or coincident with the shoot. In contrast, somatic embryos germinate in reverse sequence, with the cotyledons greening first, then shoot emergence and then, much later, if at all, the appearance of the root. Somatic seedlings, produced from the maturation medium, showed 100% survival when planted in a field setting. Somatic seedlings showed normal yearly growth relative to standard seedlings from natural seed.

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Apoptotic Effect of Pinosylvin at a High Concentration Regulated by c-Jun N-Terminal Kinase in Bovine Aortic Endothelial Cells (혈관내피세포에서 c-Jun N-terminal kinase에 의해 조절되는 세포사멸에 고농도의 피노실빈이 미치는 효과)

  • Song, Jina;Park, Jinsun;Jeong, Eunsil;So, A-Young;Pyee, Jaeho;Park, Heonyong
    • Journal of Life Science
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    • v.25 no.4
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    • pp.416-424
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    • 2015
  • Pinosylvin is a stilbenoid found in the Pinus species. Pinosylvin at ~pM to ~nM concentrations induces cell proliferation, cell migration and anti-inflammatory activity in endothelial cells. However, it was recently reported that pinosylvin at high concentrations (50 to 100 μM) induces cell death in bovine aortic endothelial cells. In this study, we conducted a series of experiments to discover how pinosylvin at a high concentration (50 μM) induces endothelial cell death. Pinosylvin at the high concentration was shown to induce endothelial cell apoptosis through enhancing caspase-3 activity, flip-flop of phosphatidyl serine, and nuclear fragmentation. We found that pinosylvin at the high concentration additively increased caspase-3 activity enhanced by serum-starvation or treatment with 100 μM etoposide. We also determined that pinosylvin at the high concentration promoted activations of c-Jun N-terminal kinase (JNK) and endothelial nitric oxide synthetase (eNOS). We further ran a series of experiments to find out which signaling molecule plays a critical role in the pinosylvin-induced apoptosis. We finally found that SP-600125, a JNK inhibitor, had an inhibitory effect on the pinosylvin-induced endothelial cell death, but L-NAME, an eNOS inhibitor, had no effect. These data indicate that JNK is involved in the pinosylvin-induced apoptosis. Collectively, pinosylvin at high doses induces cell apoptosis via JNK activation.

Responses of Native Trees Species in Korea under Elevated Carbon Dioxide Condition - Open Top Chamber Experiment (상부 개방형 온실을 이용한 대기 중 이산화탄소 농도 증가가 우리나라 자생 수종에 미치는 형태적, 생리적 영향)

  • Ryu, Daun;Bae, Jinho;Park, Juhan;Cho, Sungsik;Moon, Minkyu;Oh, Chang-Young;Kim, Hyun Seok
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.16 no.3
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    • pp.199-212
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    • 2014
  • The physiological responses of three common temperate species, Pinus densiflora, Fraxinus rhynchophylla, Sorbus alnifolia to elevated $CO_2$ was investigated using open top chambers with different $CO_2$ concentrations. Morphological (stomatal size, density and area) and physiological characteristics (maximum rates of photosynthesis, carboxylation and electron transport) were compared among trees grown under ambient, ambient ${\times}1.4$ (~550 ppm) and ambient ${\times}1.8$ (~700 ppm) $CO_2$ concentrations for last four years. Morphological responses were different among species. F. rhynchophyllar increased their stomatal size and S. alnifolia had higher stomatal density under elevated $CO_2$ than ambient. Stomatal area decreased in P. densiflora, whereas it increased in S. alnifolia. However, the maximum photosynthesis rate increased in all species up to 43.5% by S. alnifolia under elevated $CO_2$ and the enhancement increased with time. Even with four years of exposure to elevated $CO_2$, there was no sign of acclimation in the maximum carboxylation rate and the maximum electron transport rates in all species. Especially, S. alnifolia even showed the temporary increase of photosynthetic capacities in spring, when leaf nitrogen concentration was high with new leaf development. There was no significant differences in diameter growth rate in any species due to high variation in their tree sizes, however accumulated diameter and biomass for four years showed significantly increment in all species under elevated $CO_2$. For example, S. alnifolia showed 59% increase in diameter at the ambient ${\times}1.8$ (~700 ppm) compared to ambient.

Characteristics of Community Structure for Forest Vegetation on Manisan, Ganghwado (강화도 마니산 산림식생의 군집구조 특성)

  • Shin, Hak-Sub;Shin, Jae-Kwon;Kim, Hye-Jin;Han, Sang-Hak;Lee, Won-Hee;Yun, Chung-Weon
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.16 no.1
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    • pp.11-21
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    • 2014
  • The purpose of this study was to furnish basic information for forest community ecology and to accumulate vegetational datum related to hierarchy of forest community for the efficient management of forest vegetation in Mt. Mani. Samples were collected and analyzed by 32 releves from August to October in 2010 using phytosociological analysis methodology of Z-M school and importance value analysis. The results were summarized that the forest vegetation was classified into 5 units in total. Importance value at vegetation unit 1 indicated Pinus densiflora 54.31 (18.10%), Quercus mongolica 39.21 (13.07%), Carpinus coreana 37.29 (12.43%), at vegetation unit 2 Quercus mongolica 89.43 (22.23%), Rhododendron mucronulatum 57.75 (14.43%), Carpinus coreana 47.19(11.80%), at vegetation unit 3 Styrax japonica 53.97 (13.50%), Acer mono 33.60 (8.40%), Carpinus coreana 26.48 (6.62%), Quercus serrata 22.51 (5.64%), at vegetation unit 4 Carpinus coreana 47.70 (11.92%), Quercus acutissima 38.40 (9.60%) and at vegetation unit 5 Evodia daniellii 80.59 (20.14%), Robinia pseudoacacia 35.00 (8.74%), Pueraria thunbergiana 28.63 (7.15%), Quercus dentata 28.20 (7.05%) in the order, respectively.

Ecological Importance of Water Budget and Synergistic Effects of Water Stress of Plants due to Air Pollution and Soil Acidification in Korea (한국에서 수분수지의 생태적 중요성과 대기오염 및 토양 산성화로 인한 식물의 수분스트레스 증대 효과)

  • 이창석;이안나
    • The Korean Journal of Ecology
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    • v.26 no.3
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    • pp.143-150
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    • 2003
  • Korea has plentiful precipitation but rainfall events concentrate on several months of rainy season in her weather condition. Korea, therefore, experiences drought for a given period every year. Moreover the soil has usually low water holding capacity, as it is composed coarse particles originated from the granite. Response of several oaks and the Korean red pine (Pinus densiflora) on water stress showed that water budget was significant factor determining vegetation distribution. In addition, dehydration level due to cold resistance mechanism of several evergreen plants during the winter season was closely related to their distribution in natural condition. Experimental result under water stress showed that the Korean red pine was very tolerant to desiccation but the seedlings showed high mortality during the dry season. The mortality tended to proportionate to soil moisture content of each site. A comparison between soil moisture content during June when it is severe dry season and moisture content of the culture soil when the pine seedlings reached the permanent wilting point due to water withheld proved that high mortality during the dry season was due to water deficit. Water potential of sample plants measured during the exposure experiment to the air pollutant showed a probability that water related factors would dominate the occurrence of visible damage and the tolerance level of sample plants. In both field survey and laboratory experiment, plants exposed to air pollution showed more rapid transpiration than those grown in the unpolluted condition. The result would due to injury of leaf surface by air pollutants. Aluminum (Al/sup 3+/) increased in the acid soil not only inhibits root growth but also leads to abnormal distribution of root system and thereby caused water stress. The water stresses due to air pollution and soil acidification showed a possibility that they play dominating roles in inducing forest decline additionally to the existing water deficit due to weather and soil conditions in Korea. Sludge, which can contribute to improve field capacity, as it is almost composed of organic matter, showed an effect ameliorating the retarded growth of plant in the acidified soil. The effect was not less than that of dolomite known in widely as such a soil ameliorator. Litter extract contributed also to mitigate the water stress due to toxic Al/sup 3+/. We prepared a model showing the potential interaction of multiple stresses, which can cause forest decline in Korea by synthesizing those results. Furthermore, we suggested restoration plans, which can mitigate such forest decline in terms of soil amelioration and vegetation restoration.

Environmental Adaptability of Eupatorium rugosum : Relationship between Accumulation of Heavy Metals and Phenolic Compounds (서양등골나물의 환경적응력 : 중금속 축적과 Phenolic Compounds의 관계)

  • 김용옥;박종야;이호준
    • The Korean Journal of Ecology
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    • v.26 no.1
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    • pp.5-12
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    • 2003
  • Seed germination rate and seedling growth were measured on 6 different species(Phytolacca americana, Eupatorium rugosum, Rumex acetocella, Echinochloa crusgalli, Cassia mimosoides var. nomame, Setaria viridis) treated with leaf extract of E. rugosum. Total phenolic compound and heavy metal were analyzed on leaf and soil with and without E. rugosum. The growth of P. americana seedlings were stimulated by 10% and 25% of E. rugosum water extract treatment. The content of total phenolic compounds in soil was lower than that of leaf extract, and 25% was confirmed as threshold concentration in natural systems because the total phenolic compounds were not significantly different between the control soils and the soil treated with 10%, and 25% extract. Total phenolic compound concentrations of the leaf extracts were highest (1.66 mg/l) with E. rugosum grown under the Quercus forest canopy and lowest (1.09 mg/l) for the plant grown in the mixed forest edge. Leaf extracts of plants selected in different sampling sites (Forest interior, Forest edge, under Pinus Canopy and Quercus Canopy) were significant, while soil extracts were not. Seed germination of R. acetocella and S. viridis were significantly inhibited at over 50% concentrations of E. rugosum, but C. mimosoides var. nomame was not affected at any concentration. The radicle and shoot growth of the native species group were reduced two times more than those of the exotic species group by the treatment of extracts. Especially, the seed germination percentage and dry weight of E. rugosum were greater than those of the control group by treatments with extracts of 10% and 25%. Analysis of aqueous extracts from E. rugosum by HPLC identified 6 phenolic compounds: caffeic acid (460.9 mg/l), benzoic acid (109.7 mg/l), protocatechuic acid (7.3 mg/l), ρ-hydroquinone (6.0 mg/l), cinnamic acid (2.7 mg/l) and hydroquinone (0.23 mg/l). The seed germination of P. americana was also inhibited dramatically by protocatechuic acid and cinnamic acid even though the content of caffeic acid (460.9 mg/l) was the highest among analyzed phenolic compounds. The heavy metal content of soil without A. altissima was higher than that of soil with E. rugosum. Particularly, Al, Fe and Mn was considerably high and most of the heavy metal were accumulated in leaves where a high level of total phenolic compounds was found.

Environmental Condition for the Butt-Rot of Conifers by Cauliflower Mushroom (Sparassis crispa) and Wood Quality of Larix kaempferi Damaged by the Fungus (꽃송이버섯에 의한 침엽수 심재부후 발생환경 및 낙엽송 피해목의 재질 특성)

  • Park, Hyun;Oh, Deuk-Sil;Ka, Kang Hyeon;Ryu, Sung-Ryul;Park, Joo-Saeng;Hwang, Jaehong;Park, Jun-Mo
    • Journal of Korean Society of Forest Science
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    • v.98 no.1
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    • pp.16-25
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    • 2009
  • Cauliflower mushroom (Sparassis crispa) is recently recognized as a new edible and/or medicinal mushroom cultivated with conifers. By the way, the mushroom is notorious as a brown-rot fungus that causes a buttrot of larch. So, there should be a careful consideration to apply the mushroom cultivation in coniferous stand. This study was conducted to clarify the seriousness of heartwood decay on conifers such as larch by cauliflower mushroom with surveying the mushroom producing environment and to examine whether the cultivation of cauliflower mushroom produce any problem in conifer stands or not. The mushroom occurred in various coniferous stands such as Larix kaempferi, Pinus koraiensis, P. densiflora and Abies holophylla on fertile soils with adequate moisture. Soil texture of the mushroom producing site was comparatively fine compared to general forest soils; sandy loam, loam and silty loam. Soil pH ranged from 4.6 to 5.2, and organic matter contents were 4~11%, which showed relatively wide range. We could find S. crispa by a DNA technique from the wood that seemed to have no heartwood decay by naked eyes. The damaged wood showed 30% higher moisture contents than that of sound wood, while the compressive strength was 30% lowered down compared to that of sound wood. The fungus may invade conifers through the scars occurred on roots or stems, in this case spore dispersion of the mushroom takes a great role. Thus, we concluded that forest tending activities need to be applied with considering the invasion of S. crispa, and cultivation of cauliflower mushroom in forest should be attempted very carefully. By the way, we also infer that conifer stands can be nurtured without heartwood decay by S. crispa if the stand be managed in good aeration conditions by proper silvicultural practices such as sanitary thinning.

A Study on Air Temperature Reduction Effect and the Functional Improvement of Street Green Areas in Seoul, Korea (서울 도심 가로수 및 가로녹지의 기온 저감 효과와 기능 향상 연구)

  • Jung, Hee-Eun;Han, Bong-Ho;Kwak, Jeong-In
    • Journal of the Korean Institute of Landscape Architecture
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    • v.43 no.4
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    • pp.37-49
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    • 2015
  • The goal of this research is to examine air temperature changes according to tree type, plantation type, roadside green area structure, and green volume of street green area within a city. The plantation type that could be analyzed for comparison by tree type with over 3 species was 1 rows of tree+shrubs. The results of analysis of average air temperature difference between pedestrian and car streets vis-a-vis 1 row of tree+shrub in high air temperature areas were: Pinus densiflora, $1.35^{\circ}C$; Zelkova serrata, $1.84^{\circ}C$; Ginkgo biloba, $2.00^{\circ}C$; Platanus occidentalis, $2.57^{\circ}C$. This standard large wide canopy species was analyzed by the roadside to provide shade to have a significant impact on air temperature reduction. In terms of analysis of the relationship between plantation type of roadside trees and air temperature, the average air temperature difference for 1 row of tree type was $1.80^{\circ}C$; for 2 rows of trees it was $2.15^{\circ}C$. In terms of analysis of the relationship between the roadside green area structure and air temperature, for tree type, average air temperature $1.94^{\circ}C$: for tree+shrub type, average air temperature $2.49^{\circ}C$; for tree+mid-size tree+shrub type, average air temperature $2.57^{\circ}C$. That is, air temperature reduction was more effective in a multi-layer structure than a single layer structure. In the relationship analysis of green volume and air temperature reduction, the air temperature reduction effect was enlarged as there was a large amount of green volume. There was a relationship with the green volume of the road, the size of the tree and number of tree layers and a multi-layer structured form of planting. The canopy volume was large and there were a great number of rows of the tree layer and the plantation type of multi-layer structure, which is what is meant through a relationship with the green volume along the roadside. Green composition standards for air temperature reduction effects and functional improvement were proposed based on the result. For a pedestrian street width of 3m or less in the field being ideal, deciduous broadleaf trees in which the canopy volume is small and the structure of the tree+shrub type through the greatest 1m green bend were proposed. For a pedestrian street width of over 3m, deciduous broadleaf trees in which the canopy volume is large and is multi-layer planted with green bend over 1m, tree+mid-size tree+shrub type was proposed.

An Analysis of the Hail Damages to Korean Forests in 2017 by Meteorology, Species and Topography (2017년 우박에 의한 산림피해의 기상, 수종 및 지형 특성 분석)

  • Lim, Jong-Hwan;Kim, Eunsook;Lee, Bora;Kim, Sunhee;Jang, Keunchang
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.19 no.4
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    • pp.280-292
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    • 2017
  • Hail is not a frequently occurring weather event, and there are even fewer reports of hail damages to forest stands. Since the 2000s, an increase in hail incidence has been documented in Europe and the United States. In Korea, severe hails occurred in Jeollanam-do province on May 31 and in Gyeongsangbuk-do province on June 1, 2017. Hail size was ranged from 0.5 to 5.0 cm in diameter in Jeollanam-do, and from 1.5 to 3.0 cm in Gyeongsangbuk-do. This study was aimed to analyze the hail damages to forests by species and topography based on damage-categorized maps created by using drones and aerial photographs, and to analyze relationships of the damages with meteorological factors. The total damaged forest area was 1,163.1ha in Jeollanam-do, and 2,942.3ha in Gyeongsangbuk-do. Among the 'severe' damaged area 326.7ha, 91% was distributed in Jeollanam-do, and concentrated in the city of Hwasun which covers 57.2% of the total 'severe' damaged area. The most heavily damaged species was Korean red pine(Pinus densiflora S. & Z.) followed by P. rigida. Most broad-leaved trees species including oaks were recovered without any dead trees found. Liliodendron tulipifera was the most severely damaged in terms of the rate of 'severe' degree individuals which are needed to be checked whether they will die or be recovered. Cause of the death of pines was considered as the combination of physical damage caused by the hail and long-lasting drought with high air temperature that occurred before and after the hail event. No pathogens and insects were found which might have affected to tree deaths. We suggested a dieback mechanism of the pine trees damaged by hail and drought.

Basic Studies on the Location and Spatial Organization Characteristics of the Seoul Sajikdan (서울 사직단(社稷壇)의 입지(立地)와 공간구성특성(空間構成特性)에 관한 기초연구(基礎硏究))

  • Choi, Seung-Sik;Shim, Woo-Kyung;Yoo, Jong-Ho;Jeon, Hye-Won;Choi, Jong-Hee
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.30 no.1
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    • pp.146-158
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    • 2012
  • This study aims to discussion the characteristics of location and spatial organization of the Seoul Sajikdan that has significance as one of national ceremonial place in Joseon Dynasty to understand its landscape architectural characteristics. To do this, we studied in two ways; review of historical documents and analysis of the present conditions and photographing, research materials about the form of physical structure. Through these ways, the results are as follows. First, in case of construction, Sajikdan took fundamental shape in Tae-jong 16 year(1416), since then it had been changed during Imjin War, reign of Suk-jong, and Japanese Colonial Era. It took present shape through restoration of Seoul Sajikdan in 1998. Second, in case of location, Sajikdan abuts onto southeastern range of Mt. Inwang that is Wubaekho(右白虎) of inner four mountain(內四山) of Hanyang, and it has a topography of west-high-east-low type. Also its territory is up to Maedong elementary school, the office of Sajik-dong, and a Dangun shrine. Third, in case of spatial organization, Sadan and Jikdan are in harmony of Yin-Yang(陰陽) and arranged in the center. As making Yumun and Sinmun(神門) at the center of four side of Yuwon and Juwon(周垣), they show organization of the Five Elements. And subsidiary buildings are thought that its arrangement was considered functional similarity and use convenience. Finally, in case of spatial component, Danyu structures are Sadan and Jikdan that hold a Sajikdaeje(社稷大祭), Yuwon that surround two altars, and Juwon that is outer fence of Yuwon. Architectures are Sajikdan Jeongmun, Sinsil, and Anhyangcheong. Also Prunus yedoensis, Zelkova serrata, Acer palmatum, are planted, but Pinus densiflora was main species and there needs to be replanted suitably to the site.