• Title/Summary/Keyword: soil factors

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Flowering Patterns of Miscanthus Germplasms in Korea (국내 수집 억새 유전자원의 출수 특성)

  • An, Gi-Hong;Um, Kyoung-Ran;Lee, Jun-Hee;Jang, Yun-Hui;Lee, Ji-Eun;Yu, Gyeong-Dan;Cha, Young-Lok;Moon, Yun-Ho;Ahn, Jong-Woong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.4
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    • pp.510-517
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    • 2015
  • Miscanthus has been considered as the most promising bioenergy crop for lignocellulosic biomass production. In Korea, M. sacchariflorus and M. sinensis can be found easily in all regions. It is a great advantage to utilize as important species with respect to genetic and cross-breeding programs materials for creation of novel hybrids. For successful breeding programs, it is important to precisely understand the variability of flowering traits among Miscanthus species as breeding parents materials. In this study, flowering traits were observed daily in 960 germplasms of two Miscanthus species (M. sacchariflorus and M. sinensis) for growing seasons over 2 years. The flowering process was divided into three stages. ST (sprouting time) was recorded when first leaf of the plant emerged on soil. FS1 (flowering stage 1) and FS2 (flowering stage 2) were recorded when flag leaf was firstly observed, and 1 cm of panicle was showing on at least one stem, respectively. For 2013 and 2014, the latest germplasms exerted flag leaf, i.e. September 30 (DOY of FS1 164.1) and September 4 (DOY of FS1 141.0) occurred M. sacchariflorus cv. Geodae 1 and M. sacchariflorus cv. Uram collected from Southern Korea (Jeollanam-do), while Miscanthus germplasms collected from northern Korea (Gyeonggi-do) which emerged the earliest flag leaf in July and August, significantly decreased DOY. For DOY from ST to FS2, M. sacchriflorus germplasms ranged from 140 to 190 days, and 110 to 170 days for 2013 and 2014. The highest frequency showed to 160 days for 2013, and 150 days for 2014. In M. sinensis germplasms, the highest frequency showed to 180 days for 2013, and 170 days for 2014. In the results of correlation between the day of years from ST to FS2 for 2013 and 2014, M. sacchriflorus and M. sinensis showed high coefficient of correlation (0.70 and 0.89). It can be supposed that flowering characteristics of Miscanthus are largely affected by the unique phenotypic characteristic of native habitat than environmental factors of the current planted site. This study for flowering traits of Miscanthus may provides an important information in order to expedite the introduction as breeding materials for creation of new hybrid.

Bacteriocidal Effects of Ultraviolet Irradiation for Reducing Bovine Mastitis Derived from Environmental Contamination (우분뇨 유래 젖소 유방염 저감을 위한 자외선 조사 살균의 효과 규명)

  • Kim, Dong-Hyeok;Lim, Jung-Ju;Lee, Jin-Ju;Jang, Hong-Hee;Jang, Dong-Il;Lee, Seung-Joo;Lee, Hu-Jang;Min, Won-Gi;Kwon, Sun-Hong;Kim, Sang-Hun;Oh, Kwon-Young;Kim, Suk
    • Korean Journal of Environmental Agriculture
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    • v.27 no.4
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    • pp.435-440
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    • 2008
  • Bovine mastitis is an important disease causing serious economic loss in dairy production and food poison in public health. The major causative agents of bovine mastitis include Escherichia coli (E. coli), Streptococcus agalactiae (S. agalactiae), Staphylococcus aureus (S. aureus). These bacteria were found in milk and environmental condition such as feces, water, soil and so on. Recently, many cases of mastitis are derived from environmental contamination of micro-organisms, which important factors for the spread of this disease in farm. Ultraviolet irradiation (UV) has been used as disinfection for waste and water in clinical and industrial facilities. Moreover the UV irradiation has been used as useful bactericidal agents to remove bacterial biofilms in environmental condition. In this study, we determined the bacterial replication in different percentage of water content (PWC) in sterilized saw dust and feces complexes from farm, and results showed that slightly decreased growth pattern of E. coli and S. agalactiae but increased growth pattern of S. aureus in various PWC (200, 400 and 600%) until 144 h incubation. In the bacteriocidal effect of UV irradiation to bacteria in saw dust and feces complex, the results showed that bacteriocidal effect was depended on the UV irradiation time, irradiation distance and PWC. Especially the antibacterial activity of UV irratiation is stronger in low PWC (50%), long time irradiation (50 sec), and short distance (5 cm) than other condition of this study. Furthermore UV irradiation with stirring showed increased the bactericidal effect compared without stirring. These results suggested that bovine mastitis causing agents may survive long time in environmental condition especially saw dust and feces complexes in farm and can cause a various disease including mastitis. Moreover, these data can be used as basis for application and development of UV disinfection to control of bovine mastitis from environmental contaminated bacteria in dairy farm.

Trend and Further Research of Rice Quality Evaluation (쌀의 품질평가 현황과 금후 연구방향)

  • Son, Jong-Rok;Kim, Jae-Hyun;Lee, Jung-Il;Youn, Young-Hwan;Kim, Jae-Kyu;Hwang, Hung-Goo;Moon, Hun-Pal
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47
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    • pp.33-54
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    • 2002
  • Rice quality is much dependent on the pre-and post harvest management. There are many parameters which influence rice or cooked rice qualitys such as cultivars, climate, soil, harvest time, drying, milling, storage, safety, nutritive value, taste, marketing, eating, cooking conditions, and each nations' food culture. Thus, vice evaluation might not be carried out by only some parameters. Physicochemical evaluation of rice deals with amy-lose content, gelatinizing property, and its relation with taste. The amylose content of good vice in Korea is defined at 17 to 20%. Other parameters considered are as follows; ratio of protein body-1 per total protein amount in relation to taste, and oleic/linoleic acid ratio in relation to storage safety. The rice higher Mg/K ratio is considered as high quality. The optimum value is over 1.5 to 1.6. It was reported that the contents of oligosaccharide, glutamic acid or its derivatives and its proportionalities have high corelation with the taste of rice. Major aromatic compounds in rice have been known as hexanal, acetone, pentanal, butanal, octanal, and heptanal. Recently, it was found that muco-polysaccharides are solubilized during cooking. Cooked rice surface is coated by the muco-polysaccharide. The muco-polysaccharide aye contributing to the consistency and collecting free amino acids and vitamins. Thus, these parameters might be regarded as important items for quality and taste evaluation of rice. Ingredients of rice related with the taste are not confined to the total rice grain. In the internal kernel, starch is main component but nitrogen and mineral compounds are localized at the external kernel. The ingredients related with taste are contained in 91 to 86% part of the outside kernel. For safety that is considered an important evaluation item of rice quality, each residual tolerance limit for agricultural chemicals must be adopted in our country. During drying, rice quality can decline by the reasons of high drying temperature, overdrying, and rapid drying. These result in cracked grain or decolored kernel. Intrinsic enzymes react partially during the rice storage. Because of these enzymes, starch, lipid, or protein can be slowly degraded, resulting in the decline of appearance quality, occurrence of aging aroma, and increased hardness of cooked rice. Milling conditions concerned with quality are paddy quality, milling method, and milling machines. To produce high quality rice, head rice must contain over three fourths of the normal rice kernels, and broken, damaged, colored, and immature kernels must be eliminated. In addition to milling equipment, color sorter and length grader must be installed for the production of such rice. Head rice was examined using the 45 brand rices circulating in Korea, Japan, America, Australia, and China. It was found that the head rice rate of brand rice in our country was approximately 57.4% and 80-86% in foreign countries. In order to develop a rice quality evaluation system, evaluation of technics must be further developed : more detailed measure of qualities, search for taste-related components, creation and grade classification of quality evaluation factors at each management stage of treatment after harvest, evaluation of rice as food material as well as for rice cooking, and method development for simple evaluation and establishment of equation for palatability. On policy concerns, the following must be conducted : development of price discrimination in conformity to rice cultivar and grade under the basis of quality evaluation method, fixation of head rice branding, and introduction of low temperature circulation.

Optimization of a Medium for the Production of Cellulase by Bacillus subtilis NC1 Using Response Surface Methodology (반응 표면 분석법을 사용한 Bacillus subtilis NC1 유래 cellulase 생산 배지 최적화)

  • Yang, Hee-Jong;Park, Chang-Su;Yang, Ho-Yeon;Jeong, Su-Ji;Jeong, Seong-Yeop;Jeong, Do-Youn;Kang, Dae-Ook;Moon, Ja-Young;Choi, Nack-Shick
    • Journal of Life Science
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    • v.25 no.6
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    • pp.680-685
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    • 2015
  • Previously, cellulase and xylanase producing microorganism, Bacillus subtilis NC1, was isolated from soil. Based on the 16S rRNA gene sequence and API 50 CHL test the strain was identified as Bacillus subtilis, and named as B. subtilis NC1. We cloned and sequenced the genes for cellulase and xylanase. Plus, the deduced amino acid sequences from the genes of cellulase and xylanase were determined and were also identified as glycosyl hydrolases family (GH) 5 and 30, respectively. In this study to optimize the medium parameters for cellulase production by B. subtilis NC1 the RSM (response surface methodology) based on CCD (central composite design) model was performed. Three factors, tryptone, yeast extract, and NaCl, for N or C source were investigated. The cellulase activity was measured with a carboxylmethyl cellulose (CMC) plate and the 3,5-dinitrosalicylic acid (DNS) methods. The coefficient of determination (R2) for the model was 0.960, and the probability value (p=0.0001) of the regression model was highly significant. Based on the RSM, the optimum conditions for cellulase production by B. subtilis NC1 were predicted to be tryptone of 2.5%, yeast extract of 0.5%, and NaCl of 1.0%. Through the model verification, cellulase activity of Bacillus subtilis NC1 increased from 0.5 to 0.62 U/ml (24%) compared to the original medium.

Study on Adsorption Characteristics of Arsenic on Magnetite (자철석의 비소에 대한 흡착특성 연구)

  • Jeong, Hyeon-Su;Lee, Woo-Chun;Cho, Hyen-Goo;Kim, Soon-Oh
    • Journal of the Mineralogical Society of Korea
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    • v.21 no.4
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    • pp.425-434
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    • 2008
  • Arsenic contamination in soil and groundwater has recently been one of the most serious environmental concerns. This arsenic contamination can be originated from natural or anthropogenic sources. It has been well known that arsenic behavior in geo-environmental is controlled by various oxides or hydroxides, such as those of iron, manganese, and aluminum, and clay minerals. Among those, particularly, iron (oxy)hydroxides are the most effective scavengers for arsenic. For this reason, this study characterized arsenic adsorption of magnetite which is a kind of iron oxide in nature. The physicochemcial features of the magnetite were investigated to evaluate adsorption of arsenite [As(III)] and arsenate [As(V)] onto magnetite. In addition to experiments on adsorption equilibria, kinetic experiments were also conducted. The point of zero charge (PZC) and specific surface area of the laboratory-synthesized magnetite used as an arsenic adsorbent were measured 6.56 and $16.6\;g/m^2$, which values seem to be relatively smaller than those of the other iron (oxy)hydroxides. From the results of equilibria experiments, arsenite was much more adsorbed onto magnetite than arsenate, indicating the affinity of arsenite on magnetite is larger than arsenate. Arsenite and arsenate showed adsorption maxima at pHs 7 and 2, respectively. In particular, adsorption of arsenate decreased with increase in pH as a result of electrical repulsion caused by anionic arsenate and negatively-charged surface of magnetite. These results indicate that the surface charge of magnetite and the chemical speciation of arsenic should be considered as the most crucial factors in controlling arsenic. The results of kinetic experiments show that arsenate was adsorbed more quickly than arsenite and adsorption of arsenic was investigated to be mostly completed within the duration of 4 hours, regardless of chemical speciation of arsenic. When the results of kinetic experiments were fitted to a variety of kinetic models proposed so far, power function and elovich model were evaluated to be the most suitable ones which can simulate adsorption kinetics of two kinds of arsenic species onto magnetite.

Studies on Mixed-Seeding of Native Woody and Herb Species (녹화용(綠化用) 자생(自生) 목본식물(木本植物)과 초본식물종자(草本植物種子)의 혼파처리(混播處理)에 관(關)한 연구(硏究))

  • Jeon, Gi-Seong;Woo, Bo-Myeong
    • Journal of Korean Society of Forest Science
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    • v.85 no.2
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    • pp.271-279
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    • 1996
  • This study was conducted to find out the appropriate rates of mixed-seeding of native woody and herb species. It was carried out in greenhouse from 1994 to 1995. The early days germination individuals were $60no./m^2$ in case of woody species for mixed seeding treatment and the treatments were fertilization, soil surface treatment, seeding amount. The investigation factors are the number of individuals of woody and herb species, biomass, height, number of tillers, etc. The mean individuals of Lespedeza bicolor and Amorpha frutzcosa were investigated $1.14no./m^2$, $0.496no./m^2$ in 1995, and Spiraea prunifolia var. simpliciflora and Alnus hirsuta were withering to death. In 1995, the Individuals of Oenothera odorata, Themeda triandra var. japonica, Cymbopogon tortilis var. goeringii, Miscanthus sinensis, Artemisia princeps var. orientalis, Lespedeza cuneata, and Arundinella hirta were investigated $5.06no./m^2$, $1,072no./m^2$, $0no./m^2$, $412.53no./m^2$, $88.6no./m^2$, $8.9no./m^2$, $57.46no./m^2$, respectively, in case of herb species. The height and biomass of Lespedeza bicolor and Amorpha fruticosa showed very fast growth, and those of Themeda triandra var. japonica, Miscanthus sinensis had a similar tendency. For the changes in woody species, according to the seeding amount, there was no significant relationship between species, but It was highly significant in 1995. In case of the herb species, the relationship between the number of individuals and seeding amount was significant for both 1994 and 1995. The most number of tiller was Miscanthus sinensis(21), and showed Themeda triandra var. japonica, Arundinella hirta in descending order. For the mixed seeding of wood and herb species, the number of individuals, height, biomass, tiller of herb species diminished as the seeding amount of herb species increased. It can be concluded that using Lespedeza bicolor and Amorpha fruticosa for woody species and like Arundinella hirta in herb species will be efficient for revegetation measures. It seems, therefore, that the plants of revegetation methods will be used to Lespedeza bicolor, Amorpha fruticosa and Arundinella hirta, and further study is needed on the seeding amount.

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Study of Ecological Response of Endangered Sarcandra glabra (Thunb.) Nakai according to Moisture and Nutrient under Condition of Climate Change for Propagation and Restoration (증식 및 복원을 위한 기후변화조건에서 수분과 유기물에 따른 멸종위기식물 죽절초(Sarcandra glabra (Thunb.) Nakai)의 생태적 반응 연구)

  • Lee, Soo-In;Lee, Eung-Pill;Jung, Young-Ho;Kim, Eui-Ju;Lee, Jae-Keun;Lee, Seung-Yeon;Park, Jae-Hoon;Lee, Sang-Hun;You, Young-Han
    • Korean Journal of Environment and Ecology
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    • v.32 no.1
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    • pp.30-38
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    • 2018
  • The purpose of this paper is to provide reference data about propagation, restoration, and preparation of policy of endangered Sarcandra glabra (Thunb.) Nakai by investigating growth response and variation of ecological niche breadth according to moisture and nutrient under the condition of elevated $CO_2$ concentration and elevated temperature. We divided the investigation into the controlled group and treated group (elevated $CO_2$ concentration and elevated temperature) and then varied the moisture and nutrient treatment for testing. The results showed that the ecological niche breadth was wide at moisture and nutrient gradients of 0.899 and 0.844, respectively, under control. Also, the ecological niche breadth regarding the moisture and nutrient gradients under treatment simulating global warming was wider as 6.60% and 2.09%, respectively. Therefore, moisture and nutrient will not be the restriction factors concerning the growth of Sarcandra glabra under continued global warming. However, it will be advisable to specify the nutrient content condition in the soil to be 10% for population restoration when growing Sarcandra glabra in the green house which is not affected by external environment since the studies of rearing reaction reported that Sarcandra glabra prefer 10% of nutrient content than 0-5%. Furthermore, it is necessary to protect evergreen broad-leaved forest where is the natural habitat of Sarcandra glabra that has relatively high nutrient content.

The Analysis of Visiting Patterns for the Top of Seoseokdae in Mudeungsan National Park (무등산국립공원 서석대 정상부의 탐방패턴 분석)

  • Shim, Seok-Yeong;Park, Seok-Gon
    • Korean Journal of Environment and Ecology
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    • v.31 no.2
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    • pp.266-274
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    • 2017
  • The purpose of this study was to analyze the number of visitors to the top of Seoseokdae in Mudeungsan National Park, in which visitors are concentrated, and their visiting patterns, thereby suggesting measures to manage the visitors. The number of annual visitors and the numbers of regional and seasonal visitors to Mudeungsan National Park, which affect the concentration of visits to Seoseokdae were analyzed using the data produced by an automatic digitizing device. A field study was conducted to examine the number of seasonal and periodic visitors to Seoseokdae and their visiting patterns. In 2015, the number of visitors was 2,563,651 and 83.9% of the visitors visited via the Jeungsimsa and Wonhyosa area that is near Gwangju City. This area is close to the Seoseokdae area and it is easy to hike between the areas. Therefore, there was an influx of most visitors to Seoseokdae into the Jeungsimsa and Wonhyosa area. In terms of seasonal visitors, the largest number of visitors came in the fall, followed by the summer, spring, and winter in order. However, the seasonal differences were not notable. There was no statistically significant correlation between the number of visitors and meteorological factors. This result may have been because Gwangju citizens frequently visit Mudeungsan regardless of period and weather. Visitors can get to Seoseokdae via the trails into Wonhyosa and Jangbuljae. A slightly larger number of visitors used the trail into Jangbuljae in the fall and winter, whereas a larger number of visitors used the trail into Wonhyosa in the summer. In general, there is a large influx of visitors into Jangbuljae, a strategic visiting point. However, a slightly larger number of visitors may have chosen the trail into Wonhyosa in the summer because they could hike under the shade of trees. In the summer, visitors stayed in Seoseokdae for a short time with a low level of crowdedness. On the other hand, in the fall and winter, visitors stayed in the area longer because they had lunch and rested. During the time, the number of momentary maximum visitors peaked, causing extreme crowdedness. Therefore, some visitors showed the visiting pattern of entering the grassland outside the designate zone. Because this behavior can damage the grassland on the top of Seoseokdae, which can lead to soil erosion, intensive visitor management may be necessary.

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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A Study on Vascular Plants, Distribution Status and Management Plans of the Cactus Habitat (No. 429 Natural Monument) in Wolryung-ri, Jeju Island (제주 월령리 선인장군락지(천연기념물 제429호)의 관속식물상, 분포실태, 관리방안에 관한 연구)

  • Lee, Cheol-Ho;Jang, Gye-Hyun;Ryu, Tae-Bok;Choi, Byoung-Ki
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.36 no.1
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    • pp.55-66
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    • 2018
  • The cactus habitat in Jeju Island has a phytogeographically specific distribution in the East Asian region, and forms a unique landscape as the only native cactus (Opuntia ficus-indica (L.) Mill.) habitat in Korea. However, there has been no detailed investigation on the distribution of cacti in the habitat and no investigation on the diversity of the mixed composition of plants in the habitats and their correlation with the distribution of cactus populations. This study attempted to investigate the diversity of vascular plants in the Wollyeong-ri cactus habitat and record the actual distribution and trends of cactus distribution. In addition to the distribution characteristics of cacti, we also discuss the characteristics of species reflecting the characteristics of the habitat among the mixed population of plant species, as well as the biological and environmental factors that threaten the maintenance of cactus populations or require management for preservation of cacti. Considering the phenological character, we conducted the field surveys for flora identification six times between June 2015 and September 2017. The Engler classification system was used for the arrangement sequence and names of plants, and the Korean Plant Names Index was adopted for the Korean name of the species. The study results showed that the Wolryung-ri cactus habitat in Jeju Island has the characteristic physiognomy of an area dominated by cactus. For the vascular plants, a total of 125 taxa were identified, including 53 families, 104 genera, 109 species, 15 varieties and 1 forma. Endangered plants specified by the Ministry of Environment were not found. Two species, Cyrtomium falcatum and Asplenium incisum, were identified as the ferns, and no gymnosperms were found. In addition, 123 taxa of angiosperms, 91 taxa of dicotyledones and 32 taxa of monocotyledons were identified. The distributions of cacti were confirmed in 289 meshes corresponding to 59.3% of the total 487 meshes in the cactus protected area, which showed various coverage distributions ranging from 5% to 95%. Most of the meshes where no cacti were found are coastal areas with exposed basalt rocks where the soil depth has not developed or extremely restricted due to repeated waves, or areas where artificial facilities, grasslands, and observation paths have been constructed. On the other hand, there were 71 lattice points in 14.5% of the total area where the cactus showed 70% or higher dominance. Cacti are randomly distributed in these areas. They have adapted to the microhabitat environment and are found to be opportunistically distributed along the growable locations. Considering that the reproduction of cacti in the habitat is mostly dependent on parthenogenesis, the present distribution seems to reflect the potentially distributable regions of cacti in the habitat. Based on the results of field surveys, a management plan for conservation and protection of the protected areas has been proposed.