• 한국해양바이오학회지
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• 제12권1호
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• pp.50-58
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• 2020

Sphacelaria is a filamentous brown algal genus that can be epibiotic on macroalgae, marine plants, and sea turtles. Its important role in benthic ecosystems, exposure to different stressors (e.g., grazing), and use as a model organism make Sphacelaria ideal for assessing physiological responses of organisms to environmental inputs. Single-cell RNA sequencing is a powerful new probe for understanding environmental responses of organisms at the molecular (transcriptome) level, capable of delineating gene regulation in different cell types. In the case of plants, this technique requires protoplasts ("naked" plant cells). The existing protoplast isolation protocols for Sphacelaria use non-commercial enzymes and are low-yielding. This study is the first to report the production of protoplasts from Sphacelaria fusca (Hudson) S.F. Gray, using a combination of commercial enzymes, chelation, and osmolarity treatment. A simple combination of commercial enzymes (cellulase Onozuka RS, alginate lyase, and driselase) with chelation pretreatment and an increased osmolarity (2512 mOsm/L H₂O) gave a protoplast yield of 15.08 ± 5.31 × 10⁴ protoplasts/g fresh weight, with all the Sphacelaria cell types represented. Driselase had no crucial effect on the protoplast isolation. However, the increased osmolarity had a highly significant and positive effect on the protoplast isolation, and chelation pretreatment was essential for optimal protoplast yield. The protocol represents a significant step forward for studies on Sphacelaria by efficiently generating protoplasts suitable for cellular studies, including single-cell RNA sequencing and expression profiling.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권3호
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• pp.192-197
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• 2005

The effects of macronutrients $(NO_3^-,\; NH_4^+\;and\;PO_4^{3-})$ on cell growth and triterpenoids production in Centella asiatica cell suspension cultures were analyzed using the Box­Behnken response surface model experimental design. In screening and optimization experiments, $PO_4^{3-}$ as a single factor significantly influenced cell growth where increasing the phosphate level from 0.1 to 2.4 or 2.6 mM, elevated cell growth from 3.9 to $14\~16g/L$. The optimum values predicted from the response surface model are 5.05mM $NH_4^+$, 15.0mM $NO_3^-$ and 2.6mM $PO_4^{3-}$, yielding 16.0g/L cell dry weight with $99\%$ fitness to the experimental data. While the $NH_4^+-NO_3^-$ interaction influenced cell growth positively in the optimization experiment, $NH_4^+$ and $NO_3^-$ as single factors; and interactions of $NO_3^--PO_4^{3-},\;NH_4^+-PO_4^{3-}$ and $NH_4^+-NO_3^-$ were all negative in the screening experiment. Cell growth and the final pH level were positively affected by $PO_4^{3-}$, but negatively affected by $NH_4^+\;and\;NH_4^+-PO_4^{3-}$ interactions. The different effects of factors and their interactions on cell growth and final pH are influenced by a broad or narrow range of macronutrient concentrations. The productions of triterpenoids however were lower than 4mg/g cell dry weight.

• Journal of Microbiology and Biotechnology
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• 제25권12호
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• pp.2116-2124
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• 2015

To understand the effects of physicochemical factors on nitrite transformation by microalgae, a lipid-rich Chlorella with high nitrite tolerance was cultured with 8 mmol/l sodium nitrite as sole nitrogen source under different conditions. The results showed that nitrite transformation was mainly dependent on the metabolic activities of algal cells rather than oxidation of nitrite by dissolved oxygen. Light intensity, temperature, pH, NaHCO₃ concentrations, and initial cell densities had significant effects on the rate of nitrite transformation. Single-factor experiments revealed that the optimum conditions for nitrite transformation were light intensity: 300 μmol/m²/s; temperature: 30℃ pH: 7-8; NaHCO₃ concentration: 2.0 g/l; and initial cell density: 0.15 g/l; and the highest nitrite transformation rate of 1.36 mmol/l/d was achieved. There was a positive correlation between nitrite transformation rate and the growth of Chlorella. The relationship between nitrite transformation rate (mg/l/d) and biomass productivity (g/l/d) could be described by the regression equation y = 61.3x (R² = 0.9665), meaning that 61.3 mg N element was assimilated by 1.0 g dry biomass on average, which indicated that the nitrite transformation is a process of consuming nitrite as nitrogen source by Chlorella. The results demonstrated that the Chlorella suspension was able to assimilate nitrite efficiently, which implied the feasibility of using flue gas for mass production of Chlorella without preliminary removal of NO_X.

• Korean Chemical Engineering Research
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• 제46권5호
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• pp.915-921
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• 2008

고분자전해질형 연료전지의 구조 및 구성품의 물성에 따른 성능 및 물이동 현상에 관해서 많은 연구가 진행되고 있다. 이들 연구는 대체적으로 연료 전지의 BOP(Balance of plant)를 포함하는 연료전지 시스템에 관한 연구 보다는 단위 전지 및 스택에 관한 연구에 국한되어 왔다. 연료전지의 시스템에 관한 연구들 또한 세부적인 연료전지 내부의 거동에 대해서는 고려하지 않고 있었다. 이는 연료전지의 상세 모델을 이용해 연료전지 시스템에 대해 접근하기 보다는 시스템의 성능 및 동특성에 대한 연구가 주를 이루었기 때문으로 생각된다. 본 연구에서는 연료전지 음극의 수소 배출가스를 재순환할 경우 연료전지 내부에서의 거동에 미치는 영향에 대해 2차원 정상상태 모델을 이용하여 분석해 보았다. 이를 위해 2차원 정상상태 모델을 개발하였고 이를 실험을 통해 검증하는 작업을 수행하였다. 시뮬레이션은 모델식의 수정이 자유롭도록 상용패키지를 사용하지 않고 직접 구성한 알고리즘을 통해 수행되었다. 이는 여러 하이브리드 자동차용 연료전지 시스템이 연료전지 배출가스의 재순환을 고려하고 있는 상황에서 연료전지 작동 조건에 관련된 정보를 제공할 수 있다는 의의를 가진다.

• 한국항만학회지
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• 제13권2호
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• pp.389-398
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• 1999

In the biological immunity, the immune system of organisms regulates the antibody and T-cells to protect the attack from the foreign materials which are virus, germ cell, and other antigens, and supports their stable state. It has similar characteristics that has the adaptation and robustness to overcome disturbances and to control the plant of engineering application. In this paper, we build a model of the T-cell regulated immune response mechanism. We have also designed an immune response controller(IRC) focusing on the T-cell regulated immune response of the biological immune system that include both a help part to control the response and a suppress part to adjust system stabilization effect. We show some computer simulation to control the vibration of building structure system with strong wind forces excitation also demonstrate the efficiency of the proposed controller for applying a practical system even with existing nonlinear terms.

• Journal of Microbiology and Biotechnology
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• 제23권8호
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• pp.1047-1054
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• 2013

Witches' broom of lime is a disease caused by Candidatus Phytoplasma aurantifolia, which represents the most significant global threat to the production of lime trees (Citrus aurantifolia). Conventional disease management strategies have shown little success, and new approaches based on genetic engineering need to be considered. The expression of recombinant antibodies and fragments thereof in plant cells is a powerful approach that can be used to suppress plant pathogens. We have developed a single-chain variable fragment antibody (scFvIMP6) against the immunodominant membrane protein (IMP) of witches' broom phytoplasma and expressed it in different plant cell compartments. We isolated scFvIMP6 from a naïve scFv phage display library and expressed it in bacteria to demonstrate its binding activity against both recombinant IMP and intact phytoplasma cells. The expression of scFvIMP6 in plants was evaluated by transferring the scFvIMP6 cDNA to plant expression vectors featuring constitutive or phloem specific promoters in cassettes with or without secretion signals, therefore causing the protein to accumulate either in the cytosol or apoplast. All constructs were transiently expressed in Nicotiana benthamiana by agroinfiltration, and antibodies of the anticipated size were detected by immunoblotting. Plant-derived scFvIMP6 was purified by affinity chromatography, and specific binding to recombinant IMP was demonstrated by enzyme-linked immunosorbent assay. Our results indicate that scFvIMP6 binds with high activity and can be used for the detection of Ca. Phytoplasma aurantifolia and is also a suitable candidate for stable expression in lime trees to suppress witches' broom of lime.

• The Plant Pathology Journal
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• 제15권1호
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• pp.72-75
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• 1999

A scab disease occurred on watermelon seedlings by the infection of Cladosporium cucumberinum. This is the first report demonstrating the scab disease on watermelon in Korea. The casual agent attacked all plant parts of the seedlings on the ground. Infection sites were initiated with sunken and dark green spots, and then suddenly developed to large lesions softened with gummy substances. When hypocotyl and leaf stalk of seedlings were infected and softened, upper parts of seedlings were brokin down, dried and eventually died. Conidiophores of the fungus were characterized by pale olivaceous brown color, and variable length of about 3-5 mm in width. Conidia were formed I long branched chains, and conidium was ellipsoidal, fusiform or subspherical single cell mostly without septum. Morphological characteristics of the fungus were almost identical to Cladosporium cucumerinum Ellis & Authur. The fungus was also pathogenic to cucumber, squash and oriental melon, suggesting that it is a common pathogen to cucurbits. However, the fungus was not pathogenic to bottle gourd.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.119.1-119
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• 2003

The glyoxysomal nature of microbodies was determined in Botryosphaeria dothidea hyphae based on morphology and in situ enzyme characteristics by transmission electron microscopy and cytochemistry. Bound by a single membrane, microbodies had a homogeneous matrix and varied in size ranging from 200 to 400 m in diameter. Microbodies had crystalline inclusion(s) which consisted of parallel arrays of fine tubules in their matrices. Microbodies and lipid globules were frequently placed in close association with each other, forming microbody-lipid globule complexes in hyphae. The cytochemical activities of catalase and malate synthase were localized in matrices of microbodies, showing intense electron-density of the organelle. In addition, the immunogold labeling detected the presence of catalase in multivesicular bodies and hyphal cell walls as well as in matrices and crystalline inclusions of microbodies, supporting the enzyme secretion through cell walls. Meanwhile, isocitrate Iyase was localized only in matrices of microbodies. These results suggest that microbodies, particularly complexed with lipid globules, in the fungal hyphae are functionally defined as glyoxysomes, where glyoxysomal enzymes are biochemically active for the glyoxylate cycle to be a metabolic pathway in gluconeogenesis. (Mycology and Fugus Diseases)

• 한국식품영양학회지
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• 제21권3호
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• pp.269-274
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• 2008

Plants have long been used as a food source in Korea. In this study, we investigated the combined immunomodulative effects of a water extract mixture of(pine needles, Sedum sarmentosum Bunge, hijkiaorme, buckwheat and Peril a leaves) on Balb/c mice $7{\sim}8$ weeks old. The mice were fed a chow diet ad libitum and the plant extract was orally administered every other day for four weeks at two different concentrations(50 and 500 mg/kg BW). After preparing the single-cell suspension, splenocyte proliferation was determined by the MTT(3-[4,5-di-methylthiazol-2-y]-2,5-diphenyl terazolium bromide) assay. After 48hrs of incubation with the mitogens(ConA or LPS) splenocyte from the mice groups administered 50 and 500 mg/kg BW of the plant extract showed a significant increased in proliferation compared to the control group. A hemolytic plague forming cell assay was used to indicate antibody production against sheep red blood cells(SRBC). The number of antibody-secreting cells T-dependent antigen. The result of this study suggest that supplementation with this plant extract may regulate immune function by increasing splenocyte proliferation and the number of plaque forming cells.

• Journal of Plant Biotechnology
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• 제46권3호
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• pp.180-188
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• 2019

옥신은 식물의 생장과 발달 과정에서 중요한 조절자로서 기능한다. 옥신 신호전달 과정은 3개의 주요 옥신 반응 전사인자인 Auxin/indole-3-acetic acid (Aux/IAA), Gretchen Hagen 3 (GH3), 그리고 small auxin up RNA (SAUR) 유전자에 의해 조절된다. 특히, Aux/IAA는 옥신 신호에 반응하여 빠르게 축적되는 수명이 짧은 핵 단백질이다. 이 실험에서 우리는 현사시 나무(Populus alba ${\times}$ P. glandulosa)로 부터 PagAux/IAA1 유전자를 분리하고 발현 특성을 분석하였다. PagAux/IAA1 cDNA는 4개의 보존된 도메인과 2개의 nuclear localization sequence (NLS)을 포함한 200개의 아미노산을 암호화하고 있다. Southern blot 분석으로 현사시나무 genome에 PagAux/IAA1 유전자가 single copy로 존재하는 것을 확인하였다. PagAux/IAA1 유전자는 잎과 꽃에서 특이적으로 발현되었다. 그리고 PagAux/IAA1 유전자는 현탁배양세포의 생장 과정에서 초기 지수생장기에 발현되었다. PagAux/IAA1 유전자의 발현을 분석한 결과, 건조와 염 스트레스 및 식물호르몬인 ABA 처리에 의해 발현이 감소된 반면 저온 스트레스, 형성층의 세포 분열 과정 그리고 식물호르몬인 GA와 JA 처리에서 발현이 증가하였다. 따라서 PagAux/IAA1 유전자가 현사시나무에서 저온 스트레스 반응뿐 아니라 생장 과정에 관여할 것으로 판단된다.