• Journal of Plant Biotechnology
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• 제43권4호
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• pp.405-410
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• 2016

Auxins are essential in plant growth and development. The auxin-stimulated elongation of plant cells has been explained by the "acid-growth theory", which was proposed forty years ago. According to this theory, the auxin activates plasma membrane $H^+-ATPase$ to induce proton extrusion into the apoplast, promoting cell expansion through the activation of cell wall-loosening proteins such as expansins. Even though accepted as the classical theory of auxin-induced cell growth for decades, the major signaling components comprising this model were unknown, until publication of recent reports. The major gap in the acid growth theory is the signaling mechanism by which auxin activates the plasma membrane $H^+-ATPase$. Recent genetic, molecular, and biochemical approaches reveal that several auxin-related molecules, such as TIR1/AFB AUX/IAA coreceptors and SMALL AUXIN UP RNA (SAUR), serve as important components of the acid-growth model, phosphorylating and subsequently activating the plasma membrane $H^+-ATPase$. These researches reestablish the four-decade-old theory by providing us the detailed signaling mechanism of auxininduced cell growth. In this review, we discuss the recent research progress in auxin-induced cell elongation, and a set of possible future works based on the reestablished acid-growth model.

• 한국환경과학회지
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• 제23권5호
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• pp.763-770
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• 2014

The effect of $CO_2$ on the opening of stomata in the intact leaf of Commelina communis has been investigated. Full opening of stomatal apertures(around $18{\mu}m$) was achieved in the intact leaf by addition of $CO_2$($900{\mu}mol\;mol^{-1}$). At 90 minutes, the stomatal apertures of leaves treated with $CO_2$ free air were reduced. In contrast, stomata opened most widely with the treatment of $CO_2$ air at 90 minutes. The effects of light, $CO_2$ air and $CO_2$ free air on the change of membrane potential difference(PD) were measured. Fast hyperpolarization of guard cell membrane PD was recorded reaching up to -12 mV in response to light. If $CO_2$ free air was given firstly, there was no response. When light was given after $CO_2$ free air, the light effect was very clear. At the onset of $CO_2$ air, the PD showed a dramatic hyperpolarization to about -25 mV. Changes in the pH of apoplast in intact leaves in response to $CO_2$ air were observed. $CO_2$ air caused a change of 0.4 pH unit. Therefore, it can be hypothesized that $CO_2$ flowing could stimulate proton efflux which is a necessary precursor of stomatal opening.

• Journal of Plant Biology
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• 제34권2호
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• pp.129-136
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• 1991

기주식물 (토끼풀, Trifolium repens L.)의 유조직에 침입하여 생장하는 실새삼(Cuscuta australis R. Brown)의 흡기세포들의 미세구조를 조사하였다. 기주세포들과 직접 접촉되어 있는 흡기의 정면부위는 아직 분지되지 않은 선단세포들 및 이들로부터 분지하여 신장된 세포들(hyphae)로 구성되었다. 이 두 유형의 세포들은 전자밀도가 높은 세포질을 지니며, 또한 핵막이 심하게 만입된 커다란 핵을 갖는 특징을 보였다. 어떤 선단세포에서는 비후된 세포벽 물질이 기주세포벽의 중엽(middle lamellae)으로 침입하고, 파괴된 기주세포의 잔유물을 내포하는 양상을 보였다. 두 유형의 세포들은 원형질막과 세포벽이 안쪽으로 돌출하는 구조를 갖는데, 이는 기주세포들로부터 물질흡수를 촉진하기 위한 표면적의 증가현상으로 해석된다. 기주와 흡기세포의 세포벽사이를 통과하는 원형질연락사는 관찰되지 않았다. 두 식물세포의 경계면에서는 융합된 세포벽을 관찰할 수 있는데, 이 구조는 기주로부터 흡기로의 수분 및 영양물질의 수송 경로로써 작용할 수 있을 것으로 사료된다.

• 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.

• Mycobiology
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• 제36권4호
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• pp.236-241
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• 2008

The colonization of an arbuscular mycorrhizal fungus Glomus intraradices BEG110 in the soil caused a decrease in disease severity in cucumber plants after fungal inoculation with Colletotrichum orbiculare. In order to illustrate the resistance mechanism mediated by G. intraradices BEG110, infection patterns caused by C. orbiculare in the leaves of cucumber plants and the host cellular responses were characterized. These properties were characterized using transmission electron microscopy on the leaves of cucumber plants grown in soil colonized with G. intraradices BEG110. In the untreated plants, inter- and intra-cellular fungal hyphae were observed throughout the leaf tissues during both the biotrophic and necrotrophic phases of infection. The cytoplasm of fungal hyphae appeared intact during the biotrophic phase, suggesting no defense response against the fungus. However, several typical resistance responses were observed in the plants when treated with G. intraradices BEG110 including the formation of sheaths around the intracellular hyphae or a thickening of host cell walls. These observations suggest that the resistance mediated by G. intraradices BEG110 most often occurs in the symplast of the host cells rather than in the apoplast. In addition, this resistance is similar to those mediated by biotic inducers such as plant growth promoting rhizobacteria.

• The Plant Pathology Journal
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• 제29권3호
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• pp.350-355
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• 2013

Plants protect themselves from diverse potential pathogens by induction of the immune systems such as systemic acquired resistance (SAR). Most bacterial plant pathogens thrive in the intercellular space (apoplast) of plant tissues and cause symptoms. The apoplastic leaf exudate (LE) is believed to contain nutrients to provide food resource for phytopathogenic bacteria to survive and to bring harmful phytocompounds to protect plants against bacterial pathogens. In this study, we employed the pepper-Xanthomonas axonopodis system to assess whether apoplastic fluid from LE in pepper affects the fitness of X. axonopodis during the induction of SAR. The LE was extracted from pepper leaves 7 days after soil drench-application of a chemical trigger, benzothiadiazole (BTH). Elicitation of plant immunity was confirmed by significant up-regulation of four genes, CaPR1, CaPR4, CaPR9, and CaCHI2, by BTH treatment. Bacterial fitness was evaluated by measuring growth rate during cultivation with LE from BTH- or water-treated leaves. LE from BTH-treatment significantly inhibited bacterial growth when compared to that from the water-treated control. The antibacterial activity of LE from BTH-treated samples was not affected by heating at $100^{\circ}C$ for 30 min. Although the antibacterial molecules were not precisely identified, the data suggest that small (less than 5 kDa), heat-stable compound(s) that are present in BTH-induced LE directly attenuate bacterial growth during the elicitation of plant immunity.

• Molecules and Cells
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• 제46권6호
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• pp.329-336
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• 2023

Reactive oxygen species (ROS) serve as secondary messengers that regulate various developmental and signal transduction processes, with ROS primarily generated by NADPH OXIDASEs (referred to as RESPIRATORY BURST OXIDASE HOMOLOGs [RBOHs] in plants). However, the types and locations of ROS produced by RBOHs are different from those expected to mediate intracellular signaling. RBOHs produce O₂^•- rather than H₂O₂ which is relatively long-lived and able to diffuse through membranes, and this production occurs outside the cell instead of in the cytoplasm, where signaling cascades occur. A widely accepted model explaining this discrepancy proposes that RBOH-produced extracellular O₂^•- is converted to H₂O₂ by superoxide dismutase and then imported by aquaporins to reach its cytoplasmic targets. However, this model does not explain how the specificity of ROS targeting is ensured while minimizing unnecessary damage during the bulk translocation of extracellular ROS (eROS). An increasing number of studies have provided clues about eROS action mechanisms, revealing various mechanisms for eROS perception in the apoplast, crosstalk between eROS and reactive nitrogen species, and the contribution of intracellular organelles to cytoplasmic ROS bursts. In this review, we summarize these recent advances, highlight the mechanisms underlying eROS action, and provide an overview of the routes by which eROS-induced changes reach the intracellular space.

• 한국균학회지
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• 제47권4호
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• pp.359-371
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• 2019

큰느타리버섯에서 철 저장과 관련된 페리틴 단백질의 발현 및 분비를 최적화하기 위해, T-Fer 벡터에 EcoRI 및 HindIII처리를 해 페리틴 유전자를 얻은 후, BamHI으로 처리된 선형의 pPEVPR1b 분비 벡터에 클로닝하여pPEVPR1b-Fer 재조합 벡터를 구축한 다음 Agrobacterium tumefaciens LBA4404 로 도입하였다. Agrobacterium tumefaciens-mediated transformation 방법에 의해 Pleurotus eryngii로 형질전환하고 kanamycin함유된 MCM 배지에서 올바른 형질전환체를 선별하였고, 단백질 발현은 SDS-PAGE 및 항원항체 반응에 의한 western blot으로 확인하였다. 페리틴 단백질의 분비 발현은 batch culture 및 20 L airlift type fermenter에서 배양 시간 및 온도와 같은 배양 조건에 의해 최적화되었다. 페리틴 생산을 위한 배양 조건은 MCM 배지에서 25℃ 및 8 일 배양에 의해 최적화되었다. 페리틴 단백질의 양은 정량적 단백질 분석에 의해 2.4 mg/g mycelium으로 측정되었다. 그러나, PR1b (32 amino acid)의 분비서열은 큰느타리버섯 내부의 peptidase에 의해 정확하게 processing되지 않았지만, 페리틴 단백질은 균사체에서 최대로 전체단백질의 24.7% 발현되었고, 배양액에서는 검출되지 않았다. 철 결합 활성은 7.5% non-denaturing gel에서 Perls' staining에 의해 확인되었으며, 다량체 페리틴(24 subunits)이 P. eryngii 균사체에서 형성되었음을 보여준다. 생물학적 활성 측정을 위하여 페리틴을 함유한 분말을 제조하여 육계의 사료 첨가제로서의 사용 가능성에 대해 시험하였으며, 결과적으로 페리틴은 육계의 성장을 촉진하고 사료 효율 및 생산 지수를 향상시키는것으로 확인되었다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.35-35
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• 2017

How do plants take up water from soils especially when water is scarce in soils? Plants have a strategy to respond to water deficit to manage water necessary for their survival and growth. Plants regulate water transport inside them. Water flows inside the plant via (i) apoplastic pathway including xylem vessel and cell wall and (ii) cell-to-cell pathway including water channels sitting in cell membrane (aquaporins). Water transport across the root and leaf is explained by a composite transport model including those pathways. Modification of the components in those pathways to change their hydraulic conductivity can regulate water uptake and management. Apoplastic barrier is modified by producing Casparian band and suberin lamellae. These structures contain suberin known to be hydrophobic. Barley roots with more suberin content from the apoplast showed lower root hydraulic conductivity. Root hydraulic conductivity was measured by a root pressure probe. Plant root builds apoplastic barrier to prevent water loss into dry soil. Water transport in plant is also regulated in the cell-to-cell pathway via aquaporin, which has received a great attention after its discovery in early 1990s. Aquaporins in plants are known to open or close to regulate water transport in response to biotic and/or abiotic stresses including water deficit. Aquaporins in a corn leaf were opened by illumination in the beginning, however, closed in response to the following leaf water potential decrease. The evidence was provided by cell hydraulic conductivity measurement using a cell pressure probe. Changing the hydraulic conductivity of plant organ such as root and leaf has an impact not only on the speed of water transport across the plant but also on the water potential inside the plant, which means plant water uptake pattern from soil could be differentiated. This was demonstrated by a computer simulation with 3-D root structure having root hydraulic conductivity information and soil. The model study indicated that the root hydraulic conductivity plays an important role to determine the water uptake from soil with suboptimal water, although soil hydraulic conductivity also interplayed.

• 생명과학회지
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• 제32권11호
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• pp.908-917
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• 2022

전신획득저항성(SAR)은 식물이 병원체 감염 이후 식물의 비감염 조직에서도 2차 감염에 대한 방어태세를 유지할 수 있는 광범위한 식물면역시스템이다. 지금까지 많은 연구를 통해 병원체 감염시 발생하는 SAR 유도인자 또는 모바일 신호들을 발견하였음에도 불구하고 SAR 초기 모바일 신호들은 명확하지 않다. 또한 SAR유도인자로 알려진 것들도 현재까지 수송경로가 명확하지 않다. 최근 연구에 따르면 SAR 모바일신호로 알려진 Azelaic acid (AzA)와 Glycerol-3-Phosphate (G3P)는 식물의 심플라스트 경로를 통해 원형질연락사를 통해 운송되는데 반하여Salicylic acid (SA)는 아포플라스트 경로를 통해 운송되는 것으로 여겨진다. 세포질 안에서 생성된 SA는 탈수소화는 원형질막의 양성자 구동력을 만들며 SA가 세포질에서 아포플라스트로 이동을 돕는 것으로 보인다. 뿐만 아니라 식물의 큐티클은 증산작용을 조절하여SA의 수송에 관여하는 것으로 여겨진다. 이러한 근거는 큐티클층이 결핍된 돌연변이 식물에서 SA의 축적이 비정상적으로 큐티클층에 존재하는 것을 통해 확인하였다. 이 논문에서는SAR에 관여하는 여러 신호인자들의 역할과 이들의 수송방법에 대해 논의한다.