• Title/Summary/Keyword: 공배양

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Optimization of in vitro lily culture system with different treatments of taurine (타우린 처리에 의한 나리 기내 식물체 생산체계 최적화)

  • Lee, Sang-Hee;Yang, Hwan-Rae;Kim, Sun Tae;Jun, Tae Hwan;Kim, Yong Chul;Kim, Jong Bo
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.484-489
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    • 2017
  • Lilies as cut flowers are one of the most popular ornamental plants in South Korea. It is necessary to develop lily cultivars with high qualities. Therefore, highly efficient propagation systems are needed following release of elite cultivars. In this study, we used taurine treatment to improve the growth conditions including shoot and bulb formation, fresh weight gain, and reduction of rooting and browning. We experimentally evaluated the effect of taurine as a growth stimulator, at concentrations of 0, 2.5, 5, 10, 15 and 20 mg/l. The results showed that 20 mg of taurine enhanced shoot formation by 85% and increased fresh weight 5.5-fold, which was higher than the approximately four-fold increase in the control. In addition, multiple bulb formation rate was increased by 80% and rooting by 82% following exposure to 20 mg/l of taurine. The efficiency of taurine treatment was higher than that of control with 50% multiple bulb formation rate and 60% rooting rate. The browning was 10.6% at 2.5 mg/l of taurine when compared with 0.8% at 20 mg/l. Taurine showed a positive effect on the overall growth of lily plants in terms of increased fresh weight, shoot formation rate, rooting, and formation of multiple bulbs, indicating that taurine can be used as an alternative to amino acids or as an antioxidant such as citrate and vitamin C in plant tissue culture.

The development of transgenic maize expressing Actinobacillus pleuropneumoniae ApxIIA gene using Agrobacterium (아그로박테리움을 이용한 Actinobacillus pleuropneumoniae ApxIIA (ApxII toxin) 유전자 발현 옥수수 형질전환체 개발)

  • Kim, Hyun-A;Yoo, Han-Sang;Yang, Moon-Sik;Kwon, Suk-Yoon;Kim, Jin-Seog;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.37 no.3
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    • pp.313-318
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    • 2010
  • To develop edible vaccines for swine, the embryogenic calli (type II) derived from HiII genotype were inoculated with A. tumefaciens strain C58C1 containing the binary vector pMYV611, 613, 616, and V621, 622 and 623 respectively. Six of those vectors carry nptII gene which confers resistance to paromomycin and apxIIA gene producing ApxII toxin which is generated in various serum types of A. pleuropneumoniae as a target gene. The 4,120 callus clones for pMYV611, 5,959 callus clones for pMYV613, 7,581 callus clones for pMYV616, 52,329 callus clones for V621, 48,948 callus clones for V622, and 56,188 callus clones for V623 were inoculated. The frequency of positive response clone was confirmed into range of 2.3% - 4.4% for each vectors by NPTII ELISA kit assay, and the selected callus clones of them were finally 3 callus clones from pMYV611 (0.07%), 4 callus clones from pMYV613 (0.07%), 2 callus clones from pMYV616 (0.03%), 51 callus clones from V621 (0.1%), 72 callus clones from V622 (0.15%), and 102 callus clones from V623 (0.18%) respectively. From the selected callus clones of each binary vector, the integration of the apxIIA gene into maize genome was detected from 2 plants of pMYV613 and 2 plants of V623 by Southern blot analysis.

Studies on the Flocculation of Algae with Metal Ions (금속이온에 의한 조류 응결에 관한 연구)

  • Park, Yeong Jae;Lee, Sang Soo;Cho, Hye Ryun
    • Journal of Korean Society of Environmental Engineers
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    • v.37 no.8
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    • pp.441-449
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    • 2015
  • Studies on the flocculation of algae using various metal ions were carried out by measurements of optical density(OD) and zeta potential. Cyanobacteria were used as algaes. Flocculation efficiencies of cyanobacteria by an addition of metal ions were determined from OD values, and the effect of metal ions was greater in the order of $Al^{3+}$>$La^{3+}$>$Ho^{3+}$>$Fe^{2+}$>$Ca^{2+}$. Especially for trivalent metal ions, percentages of metal removed from cyanobacteria solutions on flocculation were measured, showing the same order as in flocculation efficiencies. Zeta potentials of cyanobacteria alone were measured with increasing the concentration, found to be all negative voltages, and were increased with increasing the concentration. The effect of pH on zeta potential of cyanobacteria solution was investigated. Below pH 5.5, the zeta potentials were steeply decreased with increasing pH, whereas in the range of $5.5{\leq}pH{\leq}10$ they were almost constant ($-46{\pm}1mV$) even with increasing pH. At a constant concentration of cyanobacteria ($A_{730}=0.25$), an increase in concentration of metal ions caused an increase in zeta potential of cyanobacteria solution, showing that the effect was greater in the order of $Al^{3+}$>$Ho^{3+}$>$La^{3+}{\gg}Mg^{2+}{\geq}Ca^{2+}{\gg}K^+$. At a constant metal concentration, zeta potentials were measured with increasing cyanobacteria concentration, showing that zeta potentials for $K^+$, $Mg^{2+}$ and $Ca^{2+}$ ions were negligibly changed, whereas those of $Ho^{3+}$ and $La^{3+}$ ions were decreased. Moreover, the effect of $Ho^{3+}$ ion on decreasing zeta potential was smaller than that of $La^{3+}$ ion. $Al^{3+}$ ions showed quite a different behavior that with increasing cyanobacteria concentration the zeta potentials increased and decreased thereafter. Hydrolysis of $Al^{3+}$ ions caused a difficulty to investigate coagulation or flocculation of cyanobacteria by measurement of zeta potential.

Biodeodorization of Trimethylamine by Biofilter Packed with Waste Tire-Chips (폐타이어칩 충진형 바이오 필터에 의한 Trimethylamine 제거)

  • Park, Hun-Ju;Kim, Chang-Gyun
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.8
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    • pp.789-797
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    • 2008
  • This study was conducted to investigate removal characteristics of gaseous trimethylamine(TMA) through biofilter packed with waste tire-chips. The sludge in this experiment was collected from an activated sludge operated in a wastewater treatment facility treating malodorous pollutants. The nominal amount of collected sludge was inoculated through packing materials in the filter. The removal efficiencies for varying concentrations and SVs(Space velocity) were assessed based on TMA, COD$_{Cr}$, NO$_3{^-}$-N, NO$_2{^-}$-N, NH$_4{^+}$-N and EPS(Extracellular Polymeric Substances) in leachate, since biofilter had been steady-stately operated. The influent concentration of 10 ppm of TMA was removed to approximately 95% regardless of changing SV at 120 and 180 hr$^{-1}$, but it was lowered to 80 to 90% at SV 240 hr$^{-1}$. As influent concentration was gradually increased from 5 to 55 ppm, the removal efficiencies of TMA were initially high for 95% in the range of 5 to 10 ppm, but lowered to 80% for 10 to 30 ppm. As a part of kinetic study for TMA decomposition, V$_m$(maximum substrate removal rate) and $K_s$(substrate infinity coefficient) were 14.3 g$\cdot$m$^{-3}$$\cdot$h$^{-1}$ and 0.043 g$\cdot$m$^{-3}$, respectively while adapted period was shown in the range of 100 to 150 hr. Also, the EPS concentration was consistently observed from the leachate showing 100 to 200 ppm, which indicates that biofilm has been continuously formed and sustained throughout tire-chips packed reactor.

Characteristics of Biodegradation of Geosmin using BAC Attached Bacteria in Batch Bioreactor (정수처리용 생물활성탄(BAC) 부착 박테리아를 이용한 회분식 반응기에서의 Geosmin 생분해 특성)

  • Son, Hee-Jong;Jung, Chul-Woo;Choi, Young-Ik;Jang, Seong-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.7
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    • pp.699-705
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    • 2010
  • In this study, three different biological activated carbons (BACs) were prepared from activated carbons made of each coal (F400, Calgon), coconut (Samchully) and wood(Pica, Picabiol) which were run for two and half years in the pilot plant. The attached bio-film microorganisms in and on the BACs were isolated and identified. The results showed that nine different bacteria species (Chryseomonas luteola, Stenotrophomonas maltophilia, Pseudomonas vesicularis, Aeromonas hydrophila, Spingomonas paucimobilis, Agrobacterium radiobacter, Pseudomonas fluorescens, Spirillum spp., and Pasteurella haemolytica) were isolated and identified, the dominant species was Pseudomonas sp. that had occupied 56.5%. More specifically, it was observed that the populations of the microorganisms deceased in the order: Pasteurella haemolytica (18.9%) > Chryseomonas luteola (4.0%) > Agrobacterium radiobacter (3.5%) > Aeromonas hydrophila (2.0%) in and on the BACs. After isolating of 9 species of biofilm microorganisms, the growth curve for the biomass was investigated. During 24~96 hours, the biomass has the highest concentration, and activity of the biomass was the best to uptake geosmin as carbon resources. The operation temperatures for investigating the biodegradation of geosmin were set at $4^{\circ}C$ and $25^{\circ}C$. Pseudomonas vesicularis, Pseudomonas fluorescens, Agrobacterium radiobacter and Stenotrophomonas maltophilia played a maior role in removing the target compound as geosmin. However, geosmin was not biodegraded well by Chryseomonas luteola, Spingomonas paucimobilis, and Spirillum spp.. It is also interesting to evaluate kinetics of biodegradability of geosmin. The first-order rate constants for biodegradability of geosmin at $4^{\circ}C$ and $25^{\circ}C$ were $0.00006{\sim}0.0002\;hr^{-1}$ and $0.0043{\sim}0.0046\;hr^{-1}$ respectively. Higher water temperature produced better geosmin removal rates. When concentrations of geosmin increased from 10 to 10,000 ng/L, the rate constants for biodegradability of geosmin increased from 0.0003 to $0.0882\;hr^{-1}$. As described earlier, higher geosmin concentration in the reactor produced higher rate constant.

Uptake and Transformation of RDX by Perennial Plants in Poaceae Family (Amur Silver Grass and Reed Canary Grass) under Hydroponic Culture Conditions (수경재배조건에서 다년생 벼과식물(물억새 및 갈풀)에 의한 RDX 흡수 및 분해)

  • Park, Jieun;Bae, Bumhan
    • Journal of Korean Society of Environmental Engineers
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    • v.36 no.4
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    • pp.237-245
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    • 2014
  • Amur silver grass (Miscanthus sacchariflorus) and reed canary grass (Phalaris arundinacea) were selected for RDX removal experiments in hydroponic culture conditions based on vegetation survey at three shooting ranges in northern Kyunggi province. Seedling of two plants were grown in 1/4 strength Hoagland solution in quadruplicates containing 10, 20, 30, 40 mg/L RDX for 15 days along with control and blank treatments. During the 15 days of incubation, pH and RDX concentration in medium were routinely analyzed and RDX contents in the shoot and the root were determined after solvent extraction at the end of the experiments. Both plant species showed no symptoms of RDX phyto-toxicity. The pseudo first order RDX-removal constants for amur silver grass and reed canary grass were in the range of $0.0143{\sim}0.0484day^{-1}$ and $0.0971{\sim}0.1853^{-1}$, respectively. Plant biomass normalized RDX removal rates, which decreased with the increase of initial RDX concentration, were in the range of $0.27{\sim}1.01mL{\cdot}g^{-1}day^{-1}$ and $0.87{\sim}1.66mL{\cdot}g^{-1}day^{-1}$ for amur silver grass and reed canary grass, respectively. After 15 days of treatment, RDX removal from the medium decreased from 49.0% to 23.7% with increase in the initial RDX concentration in amur silver grass and 7.3% of the initial RDX remained in the plant. In reed canary grass planted medium, less than 16.8% and 5% of the initial RDX remained in the medium and in the plant, respectively. Large quantities of unidentified polar compound, which was not detected in amur silver grass, accumulated in the root and shoot of reed silver grass.

Mechanisms of Insulinotropic Effect of YHB-2017 [Genistein] Isolated from fermentation Broths of Streptomyces sp. (방선균에서 유래한 YHB-2017 [Genistein]의 인슐린 분비 촉진 작용 기전)

  • Kwag, Won-Jae;Park, You-Hoi;Park, Jun-Chul;Lee, Byung-Kyu;Kang, Yup;Choe, Tae-Boo
    • KSBB Journal
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    • v.21 no.6 s.101
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    • pp.466-473
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    • 2006
  • Impaired insulin secretion from pancreatic beta-cells in response to glucose is an important feature in the pathology of non-insulin-dependent diabetes mellitus (NIDDM). In the course of screening for useful insulin secretagogues, we have isolated and identified YHB-2017 (Genistein) as a insulin secretion potentiator from fermentation broths of our in-house microbial library. The insulinotropic activity of YHB-2017 in isolated rat pancreatic islets was exerted only at high concentration of glucose (8.3-16 mM) but not at low concentration of glucose (3.3-5.5 mM). Also, in perifusion study with isolated rat pancreatic islets, YHB-2017 stimulated insulin secretion in a time-dependent manner when YHB-2017 was added to KRB buffer containing 16 mM glucose. In the presence of $200\;{\mu}M$ diazoxide and 35 mM KCI, which stimulates maximum $Ca^{2+}$ influx independently of KATP channel, YHB-2017 enhanced KATP channel-independent insulin secretion at high concentration glucose (16 mM). To elucidate the mechanisms of the glucose-dependent potentiation effect of YHB-2017, pharmacologic inhibitors for protein kinase A, protein kinase C and calcium/calmodulin kinase II were pre-treated and then the potentiation effect of YHB-2017 on insulin secretion was investigated. Pre-treatment of H89 as a PKA inhibitor had a significant inhibitory effect on YHB-2017-induced potentiation effect. Furthermore, western immunoblotting analyses revealed that YHB-2017 increased phosphorylation of PKA substrates and cAMP response element-binding protein (CREB) under high concentration of glucose. These results demonstrated that the insulinotropic effect of YHB-2017 is mediated through PKA signal pathway and activated amplifying $K_{ATP}$ channel-independent insulin secretion pathway.

The Effect of Weight Ratio of Carbon Source to Nitrogen Source on toe Growth and the Composition of Fatty Acid of Thraustochytrium aureum ATCC 34304 (탄소원과 질소원의 무게비가 Thraustochytrium aureum ATCC 34304의 성장 및 지방산 조성 특성에 미치는 영향)

  • Kim Won-Ho;Park Seung-Hye;Song Sang-Kyu;Bae Kyung-Dong;Hur Byung-Ki
    • KSBB Journal
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    • v.20 no.4
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    • pp.266-270
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    • 2005
  • The effect of the weight ratio of carbon source to nitrogen source on the growth and the composition change of fatty acids of Thraustochytrium aureum ATCC 34304 was investigated. The cell concentration of 5 days' culture increased and then decreased according to the increase of C/N, The ratioes for the maximum biomass were unique respectively and distributed between 1 and 4 with the initial sugar concentration in the range of 5 g/L to 25 g/L. The biomass yield, $Y_{x/s}$ decreased along with the increase of C/N, but maintained a constant value 0.35 between 10 and 20. The composition of myristic acid $(C_{14:0})$, one component of the lipid synthesized by T. aureum, was not affected with the change of C/N, but the composition of palmitic acid $(C_{16:0})$ was around $20\%$ below 4 of C/N and decreased to $15\%$ according to C/N about 4. The compositions of oleic acid $(C_{18:1})$ and linoleic acid $(C_{18:2})$ increased from 0 to $20\%\;and\;7\%$ respectively. The composition of $\gamma-linoleic$ acid $(C_{18:3})$, however, reduced from $5\%\;to\;2\%$. EPA $(C_{20:5})$ and DPA $(C_{22:5})$ showed a tendency of reduction in the weight composition according to the increase of C/N, but DHA $(C_{22:6})$ had a trend maintaining an approximately constant value, around $40\%$, irrespective of the change of C/N.

Statistical Optimization of Production Medium for Enhanced Production of Itaconic Acid Biosynthesized by Fungal Cells of Aspergillus terreus (Aspergillus terreus에 의해 생합성되는 이타콘산의 생산성 증가를 위한 통계적 생산배지 최적화)

  • Jang, Yong-Man;Shin, Woo-Shik;Lee, Do-Hoon;Kim, Sang-Yong;Park, Chul-Hwan;Jeong, Yong-Seob;Chun, Gie-Taek
    • KSBB Journal
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    • v.24 no.1
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    • pp.30-40
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    • 2009
  • Statistical optimization of the production medium was carried out in order to find an optimal medium composition in itaconic acid fermentation process. Itaconic acid utilized in the manufacture of various synthetic resins is a dicarboxylic acid biosynthesized by fungal cells of Aspergillus terreus in a branch of the TCA cycle via decarboxylation of cis-aconitate. Through OFAT (one factor at a time) experiments, six components (glucose, fructose, sucrose, soluble starch, soybean meal and cottonseed flour) were found to have significant effects on itaconic production among various carbon- and nitrogen-sources. Hence, using these six factors, interactive effects were investigated via fractional factorial design, showing that the initial concentrations of sucrose and cottonseed flour should be high for enhanced production of itaconic acid. Furthermore, through full factorial design (FFD) experiments, negative effects of $KH_2PO_4$ and $MgSO_4$ on itaconic acid biosynthesis were demonstrated, when excess amounts of the each component were initially added. Based on the FFD analysis, further statistical experiments were conducted along the steepest ascent path, followed by response surface method (RSM) in order to obtain optimal concentrations of the constituent nutrients. As a result, optimized concentrations of sucrose and cottonseed flour were found to be 90.4g/L and 53.8g/L respectively, with the corresponding production level of itaconic acid to be 4.36 g/L (about 7 fold higher productivity as compared to the previous production medium). From these experimental results, it was assumed that optimum ratio of the constituent carbon (sucrose) and nitrogen (cottonseed flour) sources was one of the most important factors for the enhanced production of itaconic acid.

Characterization of Salt Tolerant Rice Mutant Lines Derived from Azetidine-2-Carboxylic Acid Resistant Cell Lines Induced by Gamma Ray Irradiation (AZCA 저항성 돌연변이 세포주로부터 선발 육성만 내염성 벼 돌연변이 계통의 특성 검정)

  • Song, Jae-Young;Kim, Dong-Sub;Lee, Geung-Joo;Lee, In-Sok;Kang, Kwon-Kyoo;Yun, Song-Joong;Kang, Si-Yong
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.61-68
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    • 2007
  • To develop rice (Oryza sativa L.) cultivars to be planted on salt-affected sites, cell lines with enhanced proline content and resistance to growth inhibition by Azetidine-2-carboxylic acid (AZCA), a proline analogue, were screened out among calli irradiated with gamma ray of 50, 70, 90, and 120 Gy. The calli had been derived from embryo culture of the cultivar Donganbyeo. Selected AZCA resistant lines that had high proline accumulation were used as sources for selection of NaCl resistant lines. To determine an optimum concentration for selection of NaCl resistant lines, Donganbyeo seeds were initially cultured on the media containing various NaCl concentrations (0 to 2.5%) for 40 days, and 1.5% NaCl concentration was determined as the optimum concentration. One hundred sixteen salt-tolerant (ST) lines were selected from bulked 20,000 seeds of the AZCA resistant $M_{3}$ seeds in the medium containing 1.5% NaCl. The putative 33 lines ($M_{4}$ generation) considered with salt-tolerance were further analyzed for salt tolerance, amino acid and ion contents, and expression patterns of the salt tolerance-related genes. Out of the 33 lines, 7 lines were confirmed to have superior salt tolerance. Based on growth comparison of the entries, the selected mutant lines exhibited greater shoot length with average 1.5 times, root length with 1.3 times, root numbers with 1.1 times, and fresh weight with 1.5 times than control. Proline contents were increased maximum 20%, 100% and 20% in the leaf, seed and callus, respectively, of the selected lines. Compared to control, amino acid contents of the mutants were 24 to 29%, 49 to 143%, 32 to 60% higher in the leaf, seed and callus, respectively. The ratio of $Na^{+}/K^{+}$ for most of the ST-lines were lower than that of control, ranging from 1.0 to 3.8 for the leaf and 11.5 to 28.5 for the root, while the control had 3.5 and 32.9 in the leaf and root, respectively. The transcription patterns for the P5CS and NHXI genes observed by RT-PCR analysis indicated that these genes were actively expressed under salt stress. The selected mutants will be useful for the development of rice cultivar resistant to salt stress.