• Title/Summary/Keyword: 식물의 구조와 기능

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High Yield Bacterial Expression and Purification of Active Cytochrome P450 p-coumarate-3-hydroxylase (C3H), the Arabidopsis Membrane Protein (대장균 시스템을 이용한 Arabidopsis 막 단백질 cytochrome P450 p-coumarate-3hydroxylase (C3H) 활성형의 과발현 및 분리정제)

  • Yang, Hee-Jung;Kim, Wan-Yeon;Yun, Young-Ju;Yoon, Ji-Won;Kwon, Tae-Woo;Youn, Hye-Sook;Youn, Bu-Hyun
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1039-1046
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    • 2009
  • The cytochrome P450s (P450s) metabolizing natural products are among the most versatile biological catalysts known in plants, but knowledge of the structural basis for their broad substrate specificity has been limited. The activity of p-coumarate 3-hydroxylase (C3H) is thought to be essential for the biosynthesis of lignin and many other phenylpropanoid pathway products in plants however, all attempts to express and purify the protein corresponding C3H gene have failed. As a result, no conditions suitable for the unambiguous assay of the enzyme are known. The detailed understanding of the mechanism and substrate-specificity of C3Hdemands a method for the production of active protein on the milligram scale. We have developed a bacterial expression and purification system for the plant C3H, which allows for the quick expression and purification of active wild-type C3H via introduction of combinational mutagenesis. The modified cytochrome P450 C3H ($C3H_{mod}$) could be purified in the absence of detergent using immobilized metal affinity chromatography and size exclusion chromatography following extraction from isolated membranes in a high salt buffer and catalytically activated. This method makes the use of isotopic labeling of C3H for NMRstudies and X-ray crystallography practical, and is also applicable to other plant cytochrome P450 proteins.

A Functional Analysis of OsCPK11, a Calcium-dependent Protein Kinase (CDPK) Gene in Rice (벼의 칼슘-의존성 단백질 카이네즈 유전자인 OsCPK11의 기능적 분석)

  • Lee, Su-Hee;Lee, Jeong-Eun;Day, Philip;Gilroy, Simon;Kim, Sung-Ha
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1233-1244
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    • 2017
  • CDPKs have pivotal roles in plant $Ca^{2+}$-mediated transduction signaling. A total of 29 CDPK genes have been identified in rice (Oryza sativa L.), but their key functions have not been completely noted. This study focused on the OsCPK11 gene, which has not been studied, to determine its functional characteristics. A study of tissue-specific expressions revealed that the OsCPK11 gene is expressed in young leaves, mature leaves and flowers of rice. An expression of the gene was also confirmed in gibberellin-treated aleurone layers of rice. Regarding the phenotypic characteristics of Tos17-inserted OsCPK11 mutants, the heights of the mutants were not distinguishable from the heights of wild type plants, but the number of caryopses and the caryopses' weights were significantly statistically different. In addition, many grains of the mutants had white belly materials in their endosperm. The cDNA of the OsCPK11 was cloned, and an OsCPK11 protein of about 60.5 kD was obtained by using a GST affinity chromatography and an SDS-PAGE. An analysis of the amino-acid sequence of the protein indicated that the OsCPK11 protein has the structural characteristics of typical CDPKs. The results provided useful information about the functions of the OsCPK11 gene and further noted the roles CDPKs have in $Ca^{2+}$-mediated signaling in plants.

Seasonal Whole-plant Carbon Balance of Phyllospadix iwatensis on the Coast of the Korean Peninsula (한반도 연안에 분포하는 새우말의 탄소수지 계절적 변동)

  • SEUNG HYEON KIM;JONG-HYEOB KIM;HYEGWANG KIM;JIN WOO KU;KI YOUNG KIM;KUN-SEOP LEE
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.29 no.1
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    • pp.28-41
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    • 2024
  • The carbon balance serves as a valuable indicator of a plant's physiological status under diverse environmental conditions. We investigated the photosynthetic and respiratory responses of the Asian surfgrass Phyllospadix iwatensis along the northeast coast of the Korean peninsula in response to changing water temperature (ranging from 5℃ to 30℃) to estimate the seasonal whole-plant carbon balance through a series of incubation experiments. The maximum gross photosynthetic rate (Pmax) showed a significant difference among the temperature treatments, while there was no significant difference in photosynthetic efficiency (α). The maximum gross photosynthetic rate of P. iwatensis reached its peaks at 20℃ treatment (101.65 μmol O2 g-1 DW h-1) but decreased rapidly at 30℃. The saturation irradiance (Ik), compensation irradiance (Ic), and respiration rate (R) of P. iwatensis exhibited significant differences among the temperature treatments. The saturation irradiance increased up to 20-25℃ (121.59-124.50 μmol photons m-2 s-1) and sharply decreased at 30℃. The compensation irradiance and respiration rate increased steadily with rising water temperature. The ratio of Pmax to R (Pmax:R ratio) was the highest at 5℃ but dramatically decreased at 30℃. The whole-plant carbon balance, calculated based on photosynthetic parameters, respiration rates, and biomass, exhibited distinct seasonal variation, increasing during winter and spring and decreasing during summer and fall, which is consistent with the highest in situ growth in spring and severely limited growth at the highest water temperature conditions. Phyllospadix iwatensis displayed a negative carbon balance during late summer, fall, and winter, but demonstrated a positive carbon balance during spring and early summer. Our findings suggest that the rising seawater temperatures associated with climate change may lead to significant alterations in the seagrass ecosystem functioning along the rocky shores of the Korean east coast.

Patterns of Tannin Accumulation in Leaves of C-4 Euphorbia maculata (C-4 Euphorbia maculata 엽육조직 내 탄닌물질의 축적 양상)

  • Kim, In-Sun
    • Applied Microscopy
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    • v.33 no.3
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    • pp.233-241
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    • 2003
  • Patterns of tannin accumulation in leaves of C-4 Euphorbia maculata have been examined using electron microscopy. Tannins, which are secondary metabolite phenolic compounds, were found to be deposited conspicuously in vacuoles of certain tissues regardless of their stage in development. However, patterns of deposit accumulation were distinguishable by their cell type during leaf differentiation. The deposits appeared most concentrated in the concentric bundle sheath cells enclosing veins, while little or no density was detected mostly in the mesophyll cells close to the epidermis. An ultrastructural study revealed that the deposits were restricted to the vacuoles at an early stage of leaf development; during which the vacuoles were almost completely filled with the tanniferous substances. The deposits themselves took different forms ranging from granules to huge globules while expanding leaf blade. As the leaf matured, the deposits accumulated either centripetally adjacent to the inner tangential tonoplast or by penetration into the cytoplasm amongst various cellular organelles, resulting in an extremely dense cytoplasm. Electron micrographs frequently showed the delineation of each organelle by the presence of dense deposits within the cytoplasm. Some large depository vacuoles filled with tannins had a corrugated appearance on the sectioned surface. The pattern and potential role of the deposits have been discussed.

Utilization of [6]-gingerol as an origin discriminant marker influencing melanin inhibitory activity relative to its content in Pinellia ternata (반하(Pinellia ternata)에서의 [6]-gingerol 함량과 멜라닌 저해 활성에 영향을 미치는 원산지 판별 마커로의 활용)

  • An, Ju Hyeon;Won, Hyo Jun;Seo, Soo-Kyung;Kim, Doo-Young;Ku, Chang-Sub;Oh, Sei-Ryang;Ryu, Hyung Won
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.323-330
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    • 2016
  • Pinellia ternata Breitenbach, the natural medicinal plant of the Araceae family, is a perennial plant originated from the East Asia, but also widely distributed in Europe and North America. Its tuber is used as traditional medicine for treatment of various diseases such as vomiting, inflammation, and traumatic injury. Pharmacological studies revealed that P. ternata possesses anticonvulsant, anti-tumor, insecticidal, and cytotoxic activities. Despite being well-known as the useful medicinal plant, there is no reliable, standardized method for origin discrimination. Ultra performance liquid chromatography-photodiode array detector and quadrupole time of flight-mass spectrometry based metabolite-profiling was applied to explore significant metabolite for origin discrimination between Korean and Chinese P. ternata. One compound was isolated from Korean P. ternata using repeated ODS column chromatography by bioactivity guided fractionation, and determined as [6]-gingerol according to the results of spectroscopic data including nuclear magnetic resonance and MS. This compound was selected as cosmeceutical biomarker by fingerprints, and it was associated to melanin inhibitory effect determining its origin authenticity. Furthermore, the calibration curve of biomarker was prepared using validated method for the comparison of content between Korean and Chinese P. ternata. This is the report to address the selection and successful validation of the discriminant metabolite for confirmation of Korean P. ternata.

The Brassica rapa Tissue-specific EST Database (배추의 조직 특이적 발현유전자 데이터베이스)

  • Yu, Hee-Ju;Park, Sin-Gi;Oh, Mi-Jin;Hwang, Hyun-Ju;Kim, Nam-Shin;Chung, Hee;Sohn, Seong-Han;Park, Beom-Seok;Mun, Jeong-Hwan
    • Horticultural Science & Technology
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    • v.29 no.6
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    • pp.633-640
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    • 2011
  • Brassica rapa is an A genome model species for Brassica crop genetics, genomics, and breeding. With the completion of sequencing the B. rapa genome, functional analysis of the genome is forthcoming issue. The expressed sequence tags are fundamental resources supporting annotation and functional analysis of the genome including identification of tissue-specific genes and promoters. As of July 2011, 147,217 ESTs from 39 cDNA libraries of B. rapa are reported in the public database. However, little information can be retrieved from the sequences due to lack of organized databases. To leverage the sequence information and to maximize the use of publicly-available EST collections, the Brassica rapa tissue-specific EST database (BrTED) is developed. BrTED includes sequence information of 23,962 unigenes assembled by StackPack program. The unigene set is used as a query unit for various analyses such as BLAST against TAIR gene model, functional annotation using MIPS and UniProt, gene ontology analysis, and prediction of tissue-specific unigene sets based on statistics test. The database is composed of two main units, EST sequence processing and information retrieving unit and tissue-specific expression profile analysis unit. Information and data in both units are tightly inter-connected to each other using a web based browsing system. RT-PCR evaluation of 29 selected unigene sets successfully amplified amplicons from the target tissues of B. rapa. BrTED provided here allows the user to identify and analyze the expression of genes of interest and aid efforts to interpret the B. rapa genome through functional genomics. In addition, it can be used as a public resource in providing reference information to study the genus Brassica and other closely related crop crucifer plants.

Inhibitory Effects of Aralia cordata Thunb Extracts on Nitric Oxide Synthesis in RAW 264.7 Macrophage Cells (독활(Aralia cordata Thunb) 추출물의 Nitric Oxide Synthesis 저해효과)

  • Kang, Chang-Ho;Koo, Ja-Ryong;So, Jae-Seong
    • Korean Journal of Food Science and Technology
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    • v.44 no.5
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    • pp.621-627
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    • 2012
  • Assessment was made of the effects of Aralia cordata Thunb (DH) on the cell proliferation, inducible nitric oxide synthase (iNOS) mRNA gene expression and nitric oxide (NO) production in RAW 264.7 macrophage cells. For the screening of anti-inflammatory activities, ethanolic extracts of 55 species of traditional herbal medicines were examined for inhibitory effects, and it was confirmed that DH possessed inhibitory effects on NO production. As a result, DH significantly decreased the production of NO and iNOS gene expression at a concentration of $250{\mu}g/mL$. The chloroformsoluble fractionates have the strongest No synthesis inhibitory effect. It is presumed that the inhibition of NO production in LPS-stimulated RAW 264.7 cells by DH components occurred via the modulation of iNOS and DH, and that the active compound from DH may be useful for therapeutic management of inflammatory-associate diseases.

Introduction of Water Quality Management in Korean Pond Ecosystems (국내 연못생태계의 수질관리)

  • Hwang, Soon-Jin;Kim, Han-Soon;Cheon, Se-Uk;Lee, Jea-An;Kim, Chang-Muk;Shin, Jae-Ki
    • Korean Journal of Ecology and Environment
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    • v.39 no.4 s.118
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    • pp.508-515
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    • 2006
  • This study was conducted from July to December 2004, in order to find measures to improve the water quality and the function of pond ecosystems which are often negatively affected by phytoplankton and suspended particle matters. Most of the time, the management of ponds in Korea does not consider the ecological concepts such as self-purification and nutrient recycling. Instead, conventional methods depend on other factors such as fish farming and the function of fountains and waterfalls. Sustainability of pond ecosystems must be studied with the highest emphasis given to water quality. Water discharges of inflow and outflow as well as balance adjustment for different factors must be thoroughly studied. There is a great need for studies on sustainability because it can be realized through the horizontal or vertical balances of an ecosystem. Our current research offers useful information to the academia and public on maintaining sustainability in terms of structure and function of the pond.

Current status of Brassica A genome analysis (Brassica A genome의 최근 연구 동향)

  • Choi, Su-Ryun;Kwon, Soo-Jin
    • Journal of Plant Biotechnology
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    • v.39 no.1
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    • pp.33-48
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    • 2012
  • As a scientific curiosity to understand the structure and the function of crops and experimental efforts to apply it to plant breeding, genetic maps have been constructed in various crops. Especially, in the case of Brassica crop, genetic mapping has been accelerated since genetic information of model plant $Arabidopsis$ was available. As a result, the whole $B.$ $rapa$ genome (A genome) sequencing has recently been done. The genome sequences offer opportunities to develop molecular markers for genetic analysis in $Brassica$ crops. RFLP markers are widely used as the basis for genetic map construction, but detection system is inefficiency. The technical efficiency and analysis speed of the PCR-based markers become more preferable for many form of $Brassica$ genome study. The massive sequence informative markers such as SSR, SNP and InDels are also available to increase the density of markers for high-resolution genetic analysis. The high density maps are invaluable resources for QTLs analysis, marker assisted selection (MAS), map-based cloning and comparative analysis within $Brassica$ as well as related crop species. Additionally, the advents of new technology, next-generation technique, have served as a momentum for molecular breeding. Here we summarize genetic and genomic resources and suggest their applications for the molecular breeding in $Brassica$ crop.

Morphological Differentiation of the Trap in Aquatic and Terrestrial Utricularia Species (서식환경에 따른 통발속(Utricularia) 포충낭의 형태 분화)

  • Lee, Kyoung-Lan;Kim, In-Sun
    • Applied Microscopy
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    • v.40 no.4
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    • pp.237-244
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    • 2010
  • Utricularia forms small, but complex carnivorous trap along the stem either in water or in soil depending upon species. The shapes and sizes of the traps, appendages, and trichomes are known to differ among aquatic, terrestrial and epiphytic species. In the present study, the morphology and microstructure of the trap in aquatic Utricularia japonica and terrestrial U. livida were examined using light and electron microscopy. The aim of this study was to compare the characteristics of trap features between the aquatic and terrestrial species. The trap was found to be comprised of a thin walled bladder with numerous capitate trichomes, two-armed bifid and four-armed quadrifid glands in both species; however, the traps of the two species were different in size, and number and morphology of the trichomes and glands. Aquatic Utricularia was chlorenchymatous with chloroplasts distributed throughout the body, whereas the terrestrial species was translucent without plastids due to an adaptation to underground habitats. Furthermore, the former differed considerably in that the traps developed antenna and appendages around the entrance area. A peculiar trap entrance was also noted in U. livida, which exhibited radiating rows of various trichomes within funnel-shaped tissue. A large number of glandular trichomes covered the entrance area and door surface with four trigger hairs each in the aquatic form but only two in the terrestrial form. The glandular trichomes near the door secreted a large amount of mucilage that temporarily composed the velum in the U. japonica, however, it was not observed in the terrestrial species. All of the aforementioned features were highly related in their structure and function during carnivorous mechanism in Utricularia. The current findings provide important data for further comparison of the different life forms within Utricularia.