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Optimization of particle gun-mediated transformation system in Cymbidium (유전자총을 이용한 형질전환 심비디움 식물체 생산체계 최적화)

  • Noh, Hee-Sun;Kim, Mi-Seon;Lee, Yu-Mi;Lee, Yi-Rae;Lee, Sang-Il;Kim, Jong-Bo
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.293-300
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    • 2011
  • This study is conducted to develop an efficient transformation system via particle bombardment with PLBs (Protocorm-like bodies) in Cymbidium. For this, pCAMBIA3301 vector which carries a herbicide-resistant bar gene and gus gene as a reporter gene was used for transformation with Cymbidium cultivars 'Youngflower ${\times}$ masako' line. To select transformants, proper concentration of herbicide, PPT (phosphinotricin), should be determined. As a result, 5 mg/l of PPT was selected as a proper concentration. Further, proper conditions for particle bombardment were determined to obtain a high frequency of transformation. Results showed that 1.0 ${\mu}g$ of DNA concentration, 1,100 and 1,350 psi for helium gas pressure, 1.0 ${\mu}m$ of gold particle and 6 cm of target distance showed the best result for the particle bombardment experiment. Also, pre-treatment with combination 0.2 M sorbitol and 0.2 M mannitol for 4 hrs prior to genetic transformation increased the transformation efficiency up to 2.5 times. Using transformation system developed in this study, 3.2 ~ 4.0 transgenic cymbidium plants can be produced from 100 bombarded PLBs on average. Putative transgenic plants produced in this system confirmed the presence of the bar gene by PCR analysis. Also, leaves from randomely selected five transgenic lines were applied for Basta solution (0.5% v/v) to check the resistance to the PPT herbicide. As a result, three of them showed resistance and one of them showed the strongest resistance with the maintenance of green color as non-transformed plants showed. Using this established transformation system, more genes of interests can be introduced into Cymbidium plants by genetic transformation in the future.

Determination of optimum gamma ray range for radiation mutagenesis and hormesis in quinoa (Chenopodium quinoa Willd.)

  • Park, Chan Young;Song, Seon Hwa;Sin, Jong Mu;Lee, Hyeon Young;Kim, Jin Baek;Shim, Sang In
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.240-240
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    • 2017
  • Quinoa (Chenopodium quinoa Willd.) is one of the ancient crops cultivated in the Andes region at an altitude of 3,500-4000m in Chile and Bolivia from 5000 BC. It contains a large amount of protein, minerals and vitamins in comparison with other crops. The cultivation area has been increasing worldwide because of its excellent resistance to various abiotic stress such as salinity, drought and low temperature. ${\gamma}$-Ray radiation of high dose is often used as a tool to induce mutations in plant breeding, but it has a deleterious effect on organisms. However, the radiation may have a positive stimulatory effect of 'hormesis' in the low dose range. This experiment was carried out to investigate the optimum dose range for creating the quinoa genetic resources and to investigate the hormesis effect at low dose on the quinoa. This experiment was performed for 120 days from November, 2016 to February, 2017 in the greenhouse of Gyeongsang National University. ${\gamma}$-Ray radiation was irradiated to seeds at 0 Gy, 50 Gy, 100 Gy, 200 Gy, 300 Gy, 400 Gy, 600 Gy, 800 Gy and 1000 Gy for 8 hours. (50 Gy) using the low level radiation facility ($Co^{60}$) of Cooperative Research Institute of Radiation Research Institute, KAERI. Fifty seeds were placed on each petri dish lined with wet filter paper and germination rate was measured at a time interval of 2 hours for 40 hrs. The length of the root length was measured one week after germination. Each treatment was carried out in 3 replicates. The growth of seedlings were investigated for 10 days after transplanting of 30 day-old seedlings. The plant height, NDVI, SPAD, Fv/Fm, and panicle weight were measured. The germination rate was highest at 50Gy and 0Gy and the rate of seeds treated with 400Gy or higher rate decreased to 25% of the seeds treated with 50Gy. The emergence rate of seedling in pot experiment was higher at the dose of 200 Gy, 300 Gy and 400 Gy than at 0 and 50Gy. However, the rate was lower at strong radiation higher than 600Gy at which $1^{st}$ leaf was not expanded fully and dead due to extreme overgrowth at 44 days after treatment (DAT). The highest value of panicle weight was observed at 50Gy (6.15g) and 100Gy (5.57g). On the other hand, the weight at high irradiated dose of 300Gy and 400Gy was decreased by about 55% compared to low dose (50 Gy). NDVI measurement also showed the highest value at 50 Gy as the growth progressed. SPAD was the highest at 400 Gy and showed positive correlation with irradiation dose except 0 Gy. Fv/Fm was high at 50 Gy up to 30 DAT and no difference between treatments was observed except for 400 Gy from 44 DAT. The plant height was the highest in 50Gy during the growing period and was higher in the order of 50Dy, 100Gy, 0Gy, 200Gy, 300Gy and 400Gy in 88 DAT. In this experiment, the optimal radiation dose for hormesis was 50Gy and 100Gy, and the optimal radiation dose for mutagenesis seems to be 400 Gy.

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Performance Evaluation of Bio-Membrane Hybrid Process for Treatment of Food Waste Leachate (음식물 침출수 청정화를 위한 파일롯 규모의 생물-분리막 복합공정의 성능 평가 연구)

  • Lee, Myung-Gu;Park, Chul-Hwan;Lee, Do-Hoon;Kim, Tak-Hyun;Lee, Byung-Hwan;Lee, Jin-Won;Kim, Sang-Yong
    • KSBB Journal
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    • v.23 no.1
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    • pp.90-95
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    • 2008
  • In this study, a combined process of sequential anaerobic-aerobic digestion (SAAD), fluidized-bed bioreactor (FBBR), and ultrafiltration (UF) for the treatment of small scale food waste leachate was developed and evaluated. The SAAD process was tested for performance and stability by subjecting leachate from food waste to a two-phase anaerobic digestion. The main process used FBBR composed of aerators for oxygen supply and fluidization, three 5 ton reaction chambers containing an aerobic mesophilic microorganism immobilized in PE (polyethylene), and a sedimentation chamber. The HRTs (hydraulic retention time) of the combined SAAD-FBBR-UF process were 30, 7, and 1 day, and the operation temperature was set to the optimal one for microbial growth. The pilot process maintained its performance even when the CODcr of input leachate fluctuated largely. During the operation, average CODcr, TKN, TP, and salt of the effluent were 1,207mg/L, 100mg/L, 50 mg/L, and 0.01 %, which corresponded to the removal efficiencies of 99.4%, 98.6%, 89.6%, and 98.5%, respectively. These results show that the developed process is able to manage high concentration leachate from food waste and remove CODcr, TKN, TP, and salt effectively.

Simultaneous estimation of fatty acids contents from soybean seeds using fourier transform infrared spectroscopy and gas chromatography by multivariate analysis (적외선 분광스펙트럼 및 기체크로마토그라피 분석 데이터의 다변량 통계분석을 이용한 대두 종자 지방산 함량예측)

  • Ahn, Myung Suk;Ji, Eun Yee;Song, Seung Yeob;Ahn, Joon Woo;Jeong, Won Joong;Min, Sung Ran;Kim, Suk Weon
    • Journal of Plant Biotechnology
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    • v.42 no.1
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    • pp.60-70
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    • 2015
  • The aim of this study was to investigate whether fourier transform infrared (FT-IR) spectroscopy can be applied to simultaneous determination of fatty acids contents in different soybean cultivars. Total 153 lines of soybean (Glycine max Merrill) were examined by FT-IR spectroscopy. Quantification of fatty acids from the soybean lines was confirmed by quantitative gas chromatography (GC) analysis. The quantitative spectral variation among different soybean lines was observed in the amide bond region ($1,700{\sim}1,500cm^{-1}$), phosphodiester groups ($1,500{\sim}1,300cm^{-1}$) and sugar region ($1,200{\sim}1,000cm^{-1}$) of FT-IR spectra. The quantitative prediction modeling of 5 individual fatty acids contents (palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid) from soybean lines were established using partial least square regression algorithm from FT-IR spectra. In cross validation, there were high correlations ($R^2{\geq}0.97$) between predicted content of 5 individual fatty acids by PLS regression modeling from FT-IR spectra and measured content by GC. In external validation, palmitic acid ($R^2=0.8002$), oleic acid ($R^2=0.8909$) and linoleic acid ($R^2=0.815$) were predicted with good accuracy, while prediction for stearic acid ($R^2=0.4598$), linolenic acid ($R^2=0.6868$) had relatively lower accuracy. These results clearly show that FT-IR spectra combined with multivariate analysis can be used to accurately predict fatty acids contents in soybean lines. Therefore, we suggest that the PLS prediction system for fatty acid contents using FT-IR analysis could be applied as a rapid and high throughput screening tool for the breeding for modified Fatty acid composition in soybean and contribute to accelerating the conventional breeding.

Development of Cyber R&D Platform on Total System Performance Assessment for a Potential HLW Repository ; Application for Development of Scenario through QA Procedures (고준위 방사성폐기물 처분 종합 성능 평가 (TSPA)를 위한 Cyber R&D Platform 개발 ; 시나리오 도출 과정에서의 품질보증 적용 사례)

  • Seo Eun-Jin;Hwang Yong-soo;Kang Chul-Hyung
    • Proceedings of the Korean Radioactive Waste Society Conference
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    • 2005.06a
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    • pp.311-318
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    • 2005
  • Transparency on the Total System Performance Assessment (TSPA) is the key issue to enhance the public acceptance for a permanent high level radioactive repository. To approve it, all performances on TSPA through Quality Assurance is necessary. The integrated Cyber R&D Platform is developed by KAERI using the T2R3 principles applicable for five major steps in R&D's. The proposed system is implemented in the web-based system so that all participants in TSPA are able to access the system. It is composed of FEAS (FEp to Assessment through Scenario development) showing systematic approach from the FEPs to Assessment methods flow chart, PAID (Performance Assessment Input Databases) showing PA(Performance Assessment) input data set in web based system and QA system receding those data. All information is integrated into Cyber R&D Platform so that every data in the system can be checked whenever necessary. For more user-friendly system, system upgrade included input data & documentation package is under development. Throughout the next phase R&D, Cyber R&D Platform will be connected with the assessment tool for TSPA so that it will be expected to search the whole information in one unified system.

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The development of conductive 10B thin film for neutron monitoring (중성자 모니터링을 위한 전도성 10B 박막 개발)

  • Lim, Chang Hwy;Kim, Jongyul;Lee, Suhyun;Jung, Yongju;Choi, Young-Hyun;Baek, Cheol-Ha;Moon, Myung-Kook
    • Journal of Radiation Protection and Research
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    • v.39 no.4
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    • pp.199-205
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    • 2014
  • In the field of neutron detections, $^3He$ gas, the so-called "the gold standard," is the most widely used material for neutron detections because of its high efficiency in neutron capturing. However, from variable causes since early 2009, $^3He$ is being depleted, which has maintained an upward pressure on its cost. For this reason, the demands for $^3He$ replacements are rising sharply. Research into neutron converting materials, which has not been used well due to a neutron detection efficiency lower than the efficiency of $^3He$, although it can be chosen for use in a neutron detector, has been highlighted again. $^{10}B$, which is one of the $^3He$ replacements, such as $BF_3$, $^6Li$, $^{10}B$, $Gd_2O_2S$, is being researched by various detector development groups owing to a number of advantages such as easy gamma-ray discrimination, non-toxicity, low cost, etc. One of the possible techniques for the detection is an indirect neutron detection method measuring secondary radiation generated by interactions between neutrons and $^{10}B$. Because of the mean free path of alpha particle from interactions that are very short in a solid material, the thickness of $^{10}B$ should be thin. Therefore, to increase the neutron detection efficiency, it is important to make a $^{10}B$ thin film. In this study, we fabricated a $^{10}B$ thin film that is about 60 um in thickness for neutron detection using well-known technology for the manufacturing of a thin electrode for use in lithium ion batteries. In addition, by performing simple physical tests on the conductivity, dispersion, adhesion, and flexibility, we confirmed that the physical characteristics of the fabricated $^{10}B$ thin film are good. Using the fabricated $^{10}B$ thin film, we made a proportional counter for neutron monitoring and measured the neutron pulse height spectrum at a neutron facility at KAERI. Furthermore, we calculated using the Monte Carlo simulation the change of neutron detection efficiency according to the number of thin film layers. In conclusion, we suggest a fabrication method of a $^{10}B$ thin film using the technology used in making a thin electrode of lithium ion batteries and made the $^{10}B$ thin film for neutron detection using suggested method.

Optimization of Hybrid Process of(Chemical Coagulation, Fenton Oxidation and Ceramic Membrane Filtration) for the Treatment of Reactive Dye Solutions (반응성 염료폐수 처리를 위한 화학응집, 펜톤산화, 세라믹 분리막 복합공정의 최적화)

  • Yang, Jeong-Mok;Park, Chul-Hwan;Lee, Byung-Hwan;Kim, Tak-Hyun;Lee, Jin-Won;Kim, Sang-Yong
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.3
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    • pp.257-264
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    • 2006
  • This study investigated the effects of hybrid process(chemical coagulation, Fenton oxidation and ceramic UF(ultrafiltration)) on COD and color removals of commercial reactive dyestuffs. In the case of chemical coagulation, the optimal concentrations of $Fe^{3+}$ coagulant for COD and color removals of RB49(reactive blue 49) and RY84(reactive yellow 84) were determined according to the different coagulant dose at the optimal pH. They were 2.78 mM(pH 7) in RB49 and 1.85 mM(pH 6) in RY84, respectively. In the case of Fenton oxidation, the optimal concentrations of $Fe^{3+}\;and\;H_2O_2$ were obtained. Optimal $[Fe^{2+}]:[H_2O_2]$ molar ratio of COD and color removals of RB49 and RY84 were 4.41:5.73 mM and 1.15:0.81 mM, respectively. In the case of ceramic UF, the flux and rejection of supernatant after Fenton oxidation were investigated. After ceramic UF for 9 hr, the average flux of RB49 and RY84 solutions were $53.4L/m^2hr\;and\;67.4L/m^2hr$ at 1 bar, respectively. In addition, the permeate flux increased and the average flux recovery were 98.5-99.9%(RB49) and 91.0-97.3%(RY84) according to adopting off-line cleaning(5% $H_2SO_4$). Finally, COD and color removals were 91.6-95.7% and 99.8% by hybrid process, respectively.

The Effect of Bamboo (Phyllostachys nigra var. henenis Strapf) Leaf Extract on Ultraviolet B-induced Skin Damages in Mouse (자외선 B 조사 마우스에서 피부손상에 대한 분죽 (Phyllostachys nigra var. henenis Strapf)잎 추출물의 효과)

  • Chae, Se-Lim;Lee, Hae-June;Moon, Chang-Jong;Kim, Jong-Choon;Bae, Chun-Sik;Kang, Seong-Soo;Jang, Jong-Sik;Jo, Sung-Kee;Kim, Sung-Ho
    • Journal of Radiation Protection and Research
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    • v.32 no.2
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    • pp.65-69
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    • 2007
  • The effects of bamboo (Phyllostachys nigra var. henenis Strapf) leaf extract (BLE) on the changes of ultraviolet (UV) light B radiation-induced apoptotic sunburn cell (SBC) and epidermal ATPase-positive dendritic cell (DC) in SKH1-hr or ICR mouse were investigated. The mice were treated with UVB ($200mJ/cm^2$) and were sacrificed 24 hours later. BLE (50 mg/kg of body weight) or vehicle (saline) was given i.p. at 36 and 12 hours before irradiation, and 30 minutes after irradiation. BLE cream (0.2%) or cream base (vehicle) was also topically treated at 24 hours and 15 minutes before irradiation, and immediately after irradiation. The skin of SKH1-hr mouse prepared from the back of untreated mice exhibited about 0.3 SBC/cm length of epidermis, and 24 hours after UV irradiation, the applied areas show an increased number of SBCs. But the frequency of UVB-induced SBC formation was significantly reduced by intraperitoneal injection (59.0%) and topical application (31.8%) of BLE extract. The numbers of DC in normal ICR mouse were $628.00{\pm}51.56\;or\;663.20{\pm}62.58\;per\;mm^2$ of ear epidermis. By 1 day after UVB treatment, the number of ATPase-positive $cells/mm^2$ were decreased by 39.0% or 27.1% in i.p. or topical application group with vehicle. The frequency of UVB ($200mJ/cm^2$)-induced DC decrease was reduced by treatment of BLE as 25.7% in i.p. group and 3.2% in topical application group compared with the irradiation control group. The results presented herein that BLE administration could reduce the extent of skin damages produced by UVB.

Development and Application of Training Program for RI-Biomics Manpower through Analysis of Educational Demands (교육수요 분석을 통한 RI-Biomics 전문인력 양성 프로그램 개발 및 적용)

  • Shin, Woo-Ho;Park, Tai-Jin;Yeom, Yu-Sun
    • Journal of The Korean Association For Science Education
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    • v.35 no.1
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    • pp.159-167
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    • 2015
  • RI-Biomics is a promising radiation convergence technology that combines radiation with bio science as new growth power technology. Many developed countries are focusing active support and constant exertion to dominate the RI-Biomics market in advance. In order to achieve global leadership in the RI-Biomics field, we need more highly advanced technologies and professional manpower. In fact, we have less manpower compared to technology we currently hold. In this study, we established a basic infrastructure to train professional manpower in the RI-Biomics field by developing/operating optimum training program through expert interviews and survey. The developed program has four organized sections to understand overall procedure of RI-Biomics. To evaluate our training program, we performed test operations with eight students who have a major related to RI-Biomics for three weeks in KARA (Seoul) and KAERI (Jung-eup). In detail, radioisotope usage and safety management were conducted for one week as basic course, RI-Biomics application technology was conducted for two weeks as professional course. To verify performance results of training program, we conducted to journal research, daily reports, and survey on participants. The results show a high level of satisfaction with training programs and continuous intention of involvement in our program. We also need to develop an intensive course to train high-quality human resources and to operate training program continuously. This training program will be used as basic materials for the development of RI-Biomics curriculum for university. Hence, we will expect that our training program contributes in training a professional manpower and develop RI-Biomics technology.

Development of Marker-free Transgenic Rice Expressing the Wheat Storage Protein, Glu-1Dy10, for Increasing Quality Processing of Bread and Noodles (빵과 면의 가공적성 증진을 위한 밀 저장단백질 Glu-1Dy10을 발현하는 마커프리 형질전환 벼 개발)

  • Park, Soo-Kwon;Shin, DongJin;Hwang, Woon-Ha;Hur, Yeon-Jae;Kim, Tae-Heon;Oh, Se-Yun;Cho, Jun-Hyun;Han, Sang-Ik;Lee, Seung-Sik;Nam, Min-Hee;Park, Dong-Soo
    • Journal of Life Science
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    • v.24 no.6
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    • pp.618-625
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    • 2014
  • Rice flour is used in many food products. However, dough made from rice lacks extensibility and elasticity, making it less suitable than wheat for many food products such as bread and noodles. The high-molecular weight glutenin subunits (HMW-GS) of wheat play a crucial role in determining the processing properties of the wheat grain. This paper describes the development of marker-free transgenic rice plants expressing a wheat Glu-Dy10 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation. Two expression cassettes, consisting of separate DNA fragments containing Glu-1Dy10 and hygromycin phosphotransferase II (HPTII) resistance genes, were introduced separately into Agrobacterium tumefaciens EHA105 for co-infection. Each EHA105 strain harboring Glu-1Dy10 or HPTII was infected into rice calli at a 3: 1 ratio of Glu-1Bx7 and HPTII. Among 290 hygromycin-resistant $T_0$ plants, we obtained 29 transgenic lines with both the Glu-1Dy10 and HPTII genes inserted into the rice genome. We reconfirmed the integration of the Glu-1Dy10 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the Glu-1Dy10 in transgenic rice seeds were examined by semi-quantitative RT-PCR and Western blot analysis. The marker-free plants containing only the Glu-1Dy10 gene were successfully screened in the $T_1$ generation.