• Title/Summary/Keyword: plant regeneration.

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Construction of a Transgenic Tobacco Expressing a Polydnaviral Cystatin (폴리드나바이러스 유래 시스타틴 유전자 발현 형질전환 담배 제작)

  • Kim, Yeongtae;Kim, Eunsung;Park, Youngjin;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.54 no.1
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    • pp.7-15
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    • 2015
  • CpBV (Cotesia plutellae bracovirus) is a polydnavirus and encodes a cystatin (CpBV-CST1) gene. Its overexpression suppresses insect immunity and alters insect developmental processes. This study aimed to construct a genetically modified (GM) tobacco to further explore the physiological function of the viral cystatin and to apply to control insect pests. To this end, the transgenic tobacco lines were screened in expression of the target gene and assessed in insecticidal activity. A recombinant vector (pBI121-CST) was prepared and used to transform a bacterium, Agrobacterium tumefasciens. The transformed bacteria were used to generate transgenic tobacco lines, which were induced to grow callus and resulted in about 92% of shoot regeneration. The regenerated plants were screened by PCR analysis to confirm the insertion of the target gene in the plant genome. In addition, the expression of the target gene was assessed in the regenerated plants by quantitative real-time PCR (qRT-PCR). The qRT-PCR analysis showed that the transgenic line plant expressed the target gene about 17 times more than the control tobacco, indicating a stable insertion and expression of the target gene in the transgenic tobacco line. The insecticidal activity was then analyzed using the screened transgenic tobacco lines against the teneral 1st instar larvae of the oriental tobacco budworm, Helicoverpa assulta. Though there was a variation in the insecticidal efficacy among transgenic lines, T9 and T12 lines exhibited more than 95% mortality at 7 days after feeding treatment. These results suggest that CpBV-CST1 is a useful genetic resource to be used to generate GM crop against insect pests.

Production of doubled haploid population derived from the microspore culture of rapeseed (Brassica napus L.) F1 generation and analysis of fatty acid composition (유채 잡종 1세대의 소포자 배양에 의한 배가반수체 집단 선발 및 지방산 조성 분석)

  • Lee, Ji Eun;Park, Ju Hyun;Kim, Kwang Soo;An, Da Hee;Cha, Young Lok
    • Journal of Plant Biotechnology
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    • v.49 no.1
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    • pp.74-81
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    • 2022
  • Brassica napus, an oil crop that produces rapeseed oil, is an allotetraploid (AACC, 2n = 38) produced by natural hybridization between B. rapa and B. oleracea. In this study, microspore was cultured using the F1 developed from a cross between 'EMS26' line with high oleic acid content and 'J8634-B-30' lines. The flower bud size showing the nuclear development at the late uninucleate and binucleate stage with high embryogenesis rate was 2.6 ~ 3.5 mm. Microspores were cultured using only this size and after then most microspore embryo developed into secondary embryos and then regeneration plants obtained from the developed multilobe. The analysis of the ploidy of the plants revealed that 66.7% and 27.8% of the total lines were tetraploids and octoploids, respectively. The sizes of stomatal cells in tetraploids, octoploids, and diploids were 25.5, 35.6, and 19.9 ㎛, respectively, indicating that ploidy level was positively correlated with cell size. Furthermore, 62 tetraploid doubled haploid (DH) lines were selected. The average oleic acid (C18:1) and linolenic acid (C18:3) concentrations of DH were 72.3% and 6.2%, respectively. Oleic acid and linolenic acid concentrations exceeded the two parental values in 5 and 14 DH lines, respectively, suggesting that these two fatty acids had transgressive segregation. Therefore, the DH population can be utilized for the biosynthesis of unsaturated fatty acids in rapeseed and related genes. It can also be used as a breeding material for varieties with high oleic acid concentrations.

A Basic Study on the Establishment of Preservation and Management for Natural Monument(No.374) Pyeongdae-ri Torreya nucifera forest of Jeju (천연기념물 제374호 제주 평대리 비자나무 숲의 보존·관리방향 설정을 위한 기초연구)

  • Lee, Won-Ho;Kim, Dong-Hyun;Kim, Jae-Ung;Oh, Hae-Sung;Choi, Byung-Ki;Lee, Jong-Sung
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.32 no.1
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    • pp.93-106
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    • 2014
  • In this study, Analyze environment of location, investigation into vegetation resources, survey management status and establish to classify the management area for Natural monument No.374 Pyengdae-ri Torreya nucifera forest. The results were as follows: First, Torreya nucifera forest is concerned about influence of development caused by utilization of land changes to agricultural region. Thus, establish to preservation management plan for preservation of prototypical and should be excluded development activity to cause the change of terrain that Gotjawal in the Torreya nucifera forest is factor of base for generating species diversity. Secondly, Torreya nucifera forest summarized as 402 taxa composed 91 familly 263 genus, 353 species, 41 varieties and 8 forms. The distribution of plants for the first grade & second grade appear of endangered plant to Ministry of Environment specify. But, critically endangered in forest by changes in habitat, diseases and illegal overcatching. Therefore, when establishing forest management plan should be considered for put priority on protection. Thirdly, Torreya nucifera representing the upper layer of the vegetation structure. But, old tree oriented management and conservation strategy result in poor age structure. Furthermore, desiccation of forest on artificial management and decline in Torreya nucifera habitat on ecological succession can indicate a problem in forest. Therefore, establish plan such as regulation of population density and sapling tree proliferation for sustainable characteristics of the Torreya nucifera forest. Fourth, Appear to damaged of trails caused by use. Especially, Scoria way occurs a lot of damaged and higher than the share ratio of each section. Therefore, share ratio reduction Plan should be considered through the additional development of tourism routes rather than the replacement of Scoria. Fifth, Representing high preference of the Torreya nucifera forest tourist factor confirmed the plant elements. It is sensitive to usage pressure. And requires continuous monitoring by characteristic of Non-permanent. In addition, need an additional plan such as additional development of tourism elements and active utilizing an element of high preference. Sixth, Strength of protected should be differently accordance with importance. First grade area have to maintenance of plant population and natural habitats. Set the direction of the management. Second grade areas focus on annual regeneration of the forest. Third grade area should be utilized demonstration forest or set to the area for proliferate sapling. Fourth grade areas require the introduced of partial rest system that disturbance are often found in proper vegetation. Fifth grade area appropriate to the service area for promoting tourism by utilizing natural resources in Torreya nucifera forest. Furthermore, installation of a buffer zone in relatively low ratings area and periodic monitoring to the improvement of edge effect that adjacent areas of different class.

Immobilization of Xylose Isomerase and Trial Production of High Fructose Corn Syrup (Xylose 이성화 효소의 고정화 및 이성화당의 생산)

  • Chun, Moon-Jin;Lim, Bun-Sam
    • Applied Biological Chemistry
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    • v.26 no.4
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    • pp.222-230
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    • 1983
  • This study was designed to develop a process for the immobilization of xylose isomerase(D-xylose ketol isomerase, EC 5.3.1.5) from Streptomyces griseolus previously isolated by the authors and its application on a pilot plant scale for the production of high fructose corn syrup. The biomass which has endo-excreted xylose isomerase was homogenized under a pressure of $500kg/cm^2$ and 90.8% of the enzyme recovery of the native activity was obtained as compared to 54.7% recovery by the lysozyme treatment. Ionic bonding method was adopted for the enzyme immobilization due to its many reported merits. It was found that the porous resins such as Diaion HP 20, Duolite A-7, Amberlite IRA 93 and 94 were effective in immobilizing the enzyme. In addition, it was disclosed that the regeneration form of $BO_4--$ is effective for Amberlite IRA 93 and $HCO_3-$ for Diaion HP 20. Optimal immobilization condition for Amberlite IRA 93 was pH 8.0 and $55^{\circ}C$ yielding 80.6% of immobilization. Activity decay test showed half life of the immobilized enzyme with Amberlite IRA 93 was more than 24 days at $65^{\circ}C$. The carrier was evaluated to be resuable and its result showed the relative immobilization yields were 98.2, 93.3, 90.7 and 87.5%, respectively at second, third, forth and fifth rebinding test of the enzyme on Amberlite IRA 93. Optimal temperature of the immobilized enzyme was slightly lowered and the range widened to $60\sim70^{\circ}C$, while optimal pH moved toward $8.0\sim8.3$ in its isomerization reaction. The trial production result of high fructose corn syrup in pilot scale immobilization showed that one liter of immobilized xylose isomerase (350 IXIU/ml-R) is capable producing about 293l high fructose corn syrup(75% dry substance) in 30 days.

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Growth Regulators and Colchicine Treatments for Embryo Culture Efficiency in Barley (보리 배배양 효율증진을 위한 생장조절제와 콜히친처리 효과)

  • Bong Yeon, Kim
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.6
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    • pp.757-767
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    • 1995
  • This experiment was done to determine the optimum concentration of IAA for root development in plants regenerated from the callus culture of barley embryos. Two concentrations of 2,4-D, 3ppm and 5ppm selected as an optimum among five different concentrations in the previous experiment were used for callus induction and proliferation in this experiment. For callus induction, 3ppm of 2,4-D produced 35.6% in immature embryos and 4.4% in mature embryos, while 5ppm gave 33.8% in immature and 5.6% in mature embryos. Out of 320 immature embryos cultured, 111 embryos were induced to calli and 684 plants were produced from them, while only 16 embryos were induced to calli from 320 mature embryos and 92 plants were restored. The rates of callusing and plant regeneration were 34.7%, 214% in immature embryos and 5.0%, 28.7% in mature embryos, respectively. The average root lengths and root numbers of plants restored from callus at five different IAA concentrations of 0ppm, Ippm, 5ppm, l0ppm and 30ppm were 7.9mm, 3.6; 18.4mm, 5.2; 16.1mm, 3.9; 8.5mm, 3.5 and 6.4mm, 3.4, while plants directly obtained from mature embryos were 14.8mm, 4.9; 4.9mm, 3.6; 4.3mm, 3.1; 3.6mm, 2.6 and 3.2mm, 2.1, respectively. Therefore, 1ppm gave the best result for the root. promotion in callus, whle 0ppm, a control, gave the largest root developmemt in embryos. High concentration of lAA(30ppm) in callus and any exogeneous supplement of lAA in embryos negatively affected to the root lengths and root numbers. Genotypic effect was also observed in given four varieties, Bruce, Klages, Olbori and Albori. For chromosome doubling, when 0.1% colchicine was applied on 428 plants under three different conditions such as air circulation, temperatures and growth stages, 319 plants of doubled haploids were obtained so that the rate was 74.5%

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In vitro Culture and Acclimatization of Regenerated Plants of Liliem cernum $K_{OMAROV}$ (솔나리 기내배양 및 재분화 식물체의 토양순화)

  • Kim, H.K.;Lim, Jung-Dae;Hyun, Tae-Kyung;Lee, Hyeon-Yong;Lee, Jin-Ha;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.4
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    • pp.310-317
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    • 2001
  • The regenerated-bulblets placed in liquid free media resulted in good formation of roots and bulblets. On 1/4 MS free medium, roots and bulblets were predominantly induced. The 1/4 MS liquid medium supplemented with plant growth regulators was the best suitable condition for elongation of leaves and roots. Somatic embryos were frequently developed from embryogenic callus in liquid media with 2,4-D 1mg/ l . On free liquid media, the viability of callus reduced. As the salt strength of MS media reduces, the viability of callus reduced significantly. However, Leaves were induced from several callus clumps. When leaves, roots and bulb-scale segments were placed on MS media containing NAA 1mg/ l or 2,4-D 1mg/ l and various sucrose concentration, the best result about the differentiation, growth of leaf and the differentiation of leaf was obtained on MS media added 1.5% sucrose and 2,4-D 1mg/ l, 3% sucrose and NAA 1mg/ l, and 1.5% sucrose and NAA 1mg/ l, respectively. Also the better result differentiation, growth of root and differentiation of bulb was obtained on MS media with 6% sucrose and NAA 1mg/ l. Spermidine promoted the growth of leaf and the differentiation of bulb. However, spermine promoted the differentiation of leaf, the differentiation and the growth of root in MS solid media. On the MS liquid media, both spermine and spermidine stimulated organogenesis from bulb-scale segments. Regenerated plantlets were acclimatizated and grown in greenhouse in vermiculite + perlite (1 : 1 by volume) well. The optimal soil condition of rooting for plantlets regenerated was in peat moss.

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Studies on the Mass-Propagation of Potato (Solanum tuberosum L.) by In-vitro Culture. I. Effects of Phytohormons on the Callus Induction and the Organ Differentiation from Potato Meristem tip (기내배양에 의한 감자(Solanum tuberosum L.)의 대량번식(大量繁殖)에 관한 연구(硏究) I. 몇가지 생장조절물질(生長調節物質)이 생장점(生長點)으로부터의 Callus 및 기관분화(器官分化)에 미치는 영향(影響))

  • Kim, Choong Soo;Jo, Jae Seong;Choi, Chang Yeol
    • Korean Journal of Agricultural Science
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    • v.7 no.2
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    • pp.59-64
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    • 1980
  • These experiments were carried out to define the effects of 2.4-D, NAA and Benzyladenine on the differentiation and growth of the organs and the induction of callus from the potato meristem. The results were summarized as follows ; 1. The differentiation and growth of the shoots from the potato meristem was promoted in increased concentration of Benzyladenine but the callus was not induced on the M.S. medium containing Benzyladenine. 2. On the M.S. medium containing NAA 0.5 mg/l or higher concentration of NAA, the shoots were not initiated but the callus were induced from potato meristem. The growth of callus was promoted in increased concentration of NAA. 3. The roots were initiated from 50% of potato meristems planted on the M.S. medium containing more than 0.1 mg/l of NAA but the roots were pot initiated on the medium containing 2.4-D. 4. The shoot growth was significantly increased by increasing of 2.4-D concentration up to 0.1 mg/l, but the shoots were not initiated on the medium containing 2.4-D more than 1.0 mg/l. 5. For the induction and growth of the callus from potato meristem, NAA was more effective than 2.4-D and the most effective medium was M.S. medium supplemented with 2.0 mg/l of NAA. 6. The M.S. mediums supplemented with BA 2.0 mg/l and NAA 0.1 mg/l or BA 1.0 mg/l and 2.4-D 0.1 mg/l showed good results for entire plant regeneration from potato meristem.

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Development of Intelligent ATP System Using Genetic Algorithm (유전 알고리듬을 적용한 지능형 ATP 시스템 개발)

  • Kim, Tai-Young
    • Journal of Intelligence and Information Systems
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    • v.16 no.4
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    • pp.131-145
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    • 2010
  • The framework for making a coordinated decision for large-scale facilities has become an important issue in supply chain(SC) management research. The competitive business environment requires companies to continuously search for the ways to achieve high efficiency and lower operational costs. In the areas of production/distribution planning, many researchers and practitioners have developedand evaluated the deterministic models to coordinate important and interrelated logistic decisions such as capacity management, inventory allocation, and vehicle routing. They initially have investigated the various process of SC separately and later become more interested in such problems encompassing the whole SC system. The accurate quotation of ATP(Available-To-Promise) plays a very important role in enhancing customer satisfaction and fill rate maximization. The complexity for intelligent manufacturing system, which includes all the linkages among procurement, production, and distribution, makes the accurate quotation of ATP be a quite difficult job. In addition to, many researchers assumed ATP model with integer time. However, in industry practices, integer times are very rare and the model developed using integer times is therefore approximating the real system. Various alternative models for an ATP system with time lags have been developed and evaluated. In most cases, these models have assumed that the time lags are integer multiples of a unit time grid. However, integer time lags are very rare in practices, and therefore models developed using integer time lags only approximate real systems. The differences occurring by this approximation frequently result in significant accuracy degradations. To introduce the ATP model with time lags, we first introduce the dynamic production function. Hackman and Leachman's dynamic production function in initiated research directly related to the topic of this paper. They propose a modeling framework for a system with non-integer time lags and show how to apply the framework to a variety of systems including continues time series, manufacturing resource planning and critical path method. Their formulation requires no additional variables or constraints and is capable of representing real world systems more accurately. Previously, to cope with non-integer time lags, they usually model a concerned system either by rounding lags to the nearest integers or by subdividing the time grid to make the lags become integer multiples of the grid. But each approach has a critical weakness: the first approach underestimates, potentially leading to infeasibilities or overestimates lead times, potentially resulting in excessive work-inprocesses. The second approach drastically inflates the problem size. We consider an optimized ATP system with non-integer time lag in supply chain management. We focus on a worldwide headquarter, distribution centers, and manufacturing facilities are globally networked. We develop a mixed integer programming(MIP) model for ATP process, which has the definition of required data flow. The illustrative ATP module shows the proposed system is largely affected inSCM. The system we are concerned is composed of a multiple production facility with multiple products, multiple distribution centers and multiple customers. For the system, we consider an ATP scheduling and capacity allocationproblem. In this study, we proposed the model for the ATP system in SCM using the dynamic production function considering the non-integer time lags. The model is developed under the framework suitable for the non-integer lags and, therefore, is more accurate than the models we usually encounter. We developed intelligent ATP System for this model using genetic algorithm. We focus on a capacitated production planning and capacity allocation problem, develop a mixed integer programming model, and propose an efficient heuristic procedure using an evolutionary system to solve it efficiently. This method makes it possible for the population to reach the approximate solution easily. Moreover, we designed and utilized a representation scheme that allows the proposed models to represent real variables. The proposed regeneration procedures, which evaluate each infeasible chromosome, makes the solutions converge to the optimum quickly.

Approaches on Optimum Conditions for Agrobacterium-Mediated Transformation of Phalaenopsis (호접란의 Agrobacterium 이용 형질전환 시스템의 최적조건 구명을 위한 연구)

  • Na, Ae Sil;Been, Chul Gu;Jeong, Byoung Ryong
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.1
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    • pp.1-8
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    • 2010
  • Sensitivities of PLBs of four Phalaenopsis cultivars, P. 'Taisuco Windian', P. 'Nancy Amour', P. 'Pink Twilight' and P. 'Taipei Gold' to kanamycin, spectinomycin and hygromycin at different concentrations (0, 25, 50, 100, 200, and $400mg{\cdot}L^{-1}$) were examined. Hygromycin was favorable for selecting the transformants in the genetic transformation of Phalaenopsis as PLBs of four cultivars were all dead at even $25mg{\cdot}L^{-1}$ hygromycin. Responses of PLBs of P. 'Maki Watanabe' and P. 'Brother Lawrence' to DL-phosphinothricin (PPT) were determined at different concentrations (0, 0.1. 0.25, 0.5, 1.0, 2.0, 2.5, and $5.0mg{\cdot}L^{-1}$) and $0.5mg{\cdot}L^{-1}$ PPT was thought to be suitable for selecting the transformants of Phalaenopsis. The optimum conditions for Agrobacterium cocultivation with Phalaenopsis PLBs were examined using a two-step cocultivation method in Dtps. 'City Girl' and A. tumefaciens LBA4404. In the first infection period in a 1 : 10 suspension of Agrobacterium to a VW medium, 1 hr infection showed the highest PLB survival ratio. And then, PLBs were cocultivated with a bacterial strain and a 3-day cocultivation period was better for Phalaenopsis PLBs than a prolonged period. Agrobacterium tumefaciens strains LBA4404 (pTOK233) and EHA105 (pGA643) were used to compare their efficiency on the genetic transformation of Phalaenopsis PLBs. The PLBs infected with EHA105 survived more than those infected with LBA4404 after two days in a dark condition and two weeks in light condition on a selective medium. About 1,000 PLBs for each of P. 'Maki Watanabe' and P. 'Brother Lawrence', and each bacterial strain of AGL1 (pCAMBIA3301) and LBA4404 (pTOK233) were used for the regeneration of transgenic plants. The bacterial strain AGL1 had a higher genetic transformation efficiency than LBA4404, with no significant difference between cultivars. In this study, 11 hygromycin-resistant plantlets and 32 PPT-resistant plantlets were produced, but these putative transgenic plantlets need further examinations.

A Study of Fluoride and Arsenic Adsorption from Aqueous Solution Using Alum Sludge Based Adsorbent (알럼 슬러지 기반 흡착제를 이용한 수용액상 불소 및 비소 흡착에 관한 연구)

  • Lee, Joon Hak;Ji, Won Hyun;Lee, Jin Soo;Park, Seong Sook;Choi, Kung Won;Kang, Chan Ung;Kim, Sun Joon
    • Economic and Environmental Geology
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    • v.53 no.6
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    • pp.667-675
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    • 2020
  • An Alum-sludge based adsorbent (ASBA) was synthesized by the hydrothermal treatment of alum sludge obtained from settling basin in water treatment plant. ASBA was applied to remove fluoride and arsenic in artificially-contaminated aqueous solutions and mine drainage. The mineralogical crystal structure, composition, and specific surface area of ASBA were identified. The result revealed that ASBA has irregular pores and a specific surface area of 87.25 ㎡ g-1 on its surface, which is advantageous for quick and facile adsorption. The main mineral components of the adsorbent were found to be quartz(SiO2), montmorillonite((Al,Mg)2Si4O10(OH)2·4H2O) and albite(NaAlSi3O8). The effects of pH, reaction time, initial concentration, and temperature on removal of fluoride and arsenic were examined. The results of the experiments showed that, the adsorbed amount of fluoride and arsenic gradually decreased with increasing pH. Based on the results of kinetic and isotherm experiments, the maximum adsorption capacity of fluoride and arsenic were 7.6 and 5.6 mg g-1, respectively. Developed models of fluoride and arsenic were suitable for the Langmuir and Freundlich models. Moreover, As for fluoride and arsenic, the increase rate of adsorption concentration decreased after 8 and 12 hr, respectively, after the start of the reaction. Also, the thermodynamic data showed that the amount of fluoride and arsenic adsorbed onto ASBA increased with increasing temperature from 25℃ to 35℃, indicating that the adsorption was endothermic and non-spontaneous reaction. As a result of regeneration experiments, ASBA can be regenerated by 1N of NaOH. In the actual mine drainage experiment, it was found that it has relatively high removal rates of 77% and 69%. The experimental results show ASBA is effective as an adsorbent for removal fluoride and arsenic from mine drainage, which has a small flow rate and acid/neutral pH environment.