• Title/Summary/Keyword: Plant molecular biology

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Partial Purification of OsCPK11 from Rice Seedlings and Its Biochemical Characterization (벼 유식물에서 OsCPK11의 부분 정제 및 생화학적 특성 규명)

  • Shin, Jae-Hwa;Kim, Sung-Ha
    • Journal of Life Science
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    • v.30 no.2
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    • pp.137-146
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    • 2020
  • Calcium is one of the important secondary signaling molecules in plant cells. Calcium-dependent protein kinases (CDPK)-the sensor proteins of Ca2+ and phosphorylating enzymes-are the most abundant serine/threonine kinases in plant cells. They convert and transmit signals in response to various stimuli, resulting in specific responses in plants. In rice, 31 CDPK gene families have been identified, which are mainly involved in plant growth and development and are known to play roles in response to various stress conditions. However, little is known about the biochemical characteristics of CDPK proteins. In this study, OsCPK11-a CDPK in rice-was partially purified, and its biochemical characteristics were found. Partially purified OsCPK11 from rice seedlings was obtained by three-step column chromatography that involved anion exchange chromatography consisting of DEAE, hydrophobic interaction chromatography consisting of phenyl-Sepharose, and gel filtration chromatography consisting of Sephacryl-200HR. An in vitro kinase assay using partially purified OsCPK11 was also performed. This partially purified OsCPK11 had a molecular weight of 54 kDa and showed a strong hydrophobic interaction with the hydrophobic resin. In vitro kinase assay showed that the OsCPK11 also had Ca2+-dependent autophosphorylation activity. The OsCPK11 phosphorylated histone III-S, and the optimum pH for its kinase activity was found to be 7.5~8.0. The native OsCPK11 shared several biochemical characteristics with recombinant OsCPK11 studied previously, and both had Ca2+-dependent autophosphorylation activity and favored histone III-S as a substrate for kinase activity, which also had a Ca2+-dependence.

Isolation and functional analysis of three microsomal delta-12 fatty acid desaturase genes from Camelina sativa (L.) cv. CAME (카멜리나 (Camelina sativa L. cv. CAME)로부터 3 microsomal delta-12 fatty acid desaturase 유전자들의 분리 및 기능 분석)

  • Kim, Hyojin;Go, Young Sam;Kim, Augustine Yonghwi;Lee, Sanghyeob;Kim, Kyung-Nam;Lee, Geung-Joo;Kim, Gi-Jun;Suh, Mi Chung
    • Journal of Plant Biotechnology
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    • v.41 no.3
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    • pp.146-158
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    • 2014
  • Camelina sativa that belongs to Brassicaceae family is an emerging oilseed crop. Camelina seeds contain approximately 40% storage oils per seed dry weight, which are useful for human and animal diets and industrial applications. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. In this study, three CsFAD2 genes (CsFAD2-1, CsFAD2-2 and CsFAD2-3.1) were isolated from developing seeds of Camelina sativa (L.) cv. CAME. The nucleotide and deduced amino acid sequences of three CsFAD2 genes were compared with those from dicotyledon and monocotyledon plants including Camelina cultivars Sunesone and SRS933. Three histidine motifs (HECGH, HRRHH, and HVAHH) required for FAD activity and a hydrophobic valine or isoleucine residue, which is a SNP (single nucleotide polymorphism) marker related with enzyme activity are well conserved in three CsFAD2s. The expressions of CsFAD2-1 and CsFAD2-3.1 were ubiquitously detected in various Camelina organs, whereas the CsFAD2-2 transcripts were predominantly detected in flowers and developing seeds. The contents of oleic acids decreased, whereas the amounts of linoleic acid increased in dry seeds of transgenic fad2-2 lines expressing each CsFAD2 gene compared with fad2-2 mutant, indicating that three CsFAD2 genes are functionally active. The isolated CsFAD2 genes might be applicable in metabolic engineering of storage oils with high oleic acids in oilseed crops.

A Study on Vascular Plants, Distribution Status and Management Plans of the Cactus Habitat (No. 429 Natural Monument) in Wolryung-ri, Jeju Island (제주 월령리 선인장군락지(천연기념물 제429호)의 관속식물상, 분포실태, 관리방안에 관한 연구)

  • Lee, Cheol-Ho;Jang, Gye-Hyun;Ryu, Tae-Bok;Choi, Byoung-Ki
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.36 no.1
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    • pp.55-66
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    • 2018
  • The cactus habitat in Jeju Island has a phytogeographically specific distribution in the East Asian region, and forms a unique landscape as the only native cactus (Opuntia ficus-indica (L.) Mill.) habitat in Korea. However, there has been no detailed investigation on the distribution of cacti in the habitat and no investigation on the diversity of the mixed composition of plants in the habitats and their correlation with the distribution of cactus populations. This study attempted to investigate the diversity of vascular plants in the Wollyeong-ri cactus habitat and record the actual distribution and trends of cactus distribution. In addition to the distribution characteristics of cacti, we also discuss the characteristics of species reflecting the characteristics of the habitat among the mixed population of plant species, as well as the biological and environmental factors that threaten the maintenance of cactus populations or require management for preservation of cacti. Considering the phenological character, we conducted the field surveys for flora identification six times between June 2015 and September 2017. The Engler classification system was used for the arrangement sequence and names of plants, and the Korean Plant Names Index was adopted for the Korean name of the species. The study results showed that the Wolryung-ri cactus habitat in Jeju Island has the characteristic physiognomy of an area dominated by cactus. For the vascular plants, a total of 125 taxa were identified, including 53 families, 104 genera, 109 species, 15 varieties and 1 forma. Endangered plants specified by the Ministry of Environment were not found. Two species, Cyrtomium falcatum and Asplenium incisum, were identified as the ferns, and no gymnosperms were found. In addition, 123 taxa of angiosperms, 91 taxa of dicotyledones and 32 taxa of monocotyledons were identified. The distributions of cacti were confirmed in 289 meshes corresponding to 59.3% of the total 487 meshes in the cactus protected area, which showed various coverage distributions ranging from 5% to 95%. Most of the meshes where no cacti were found are coastal areas with exposed basalt rocks where the soil depth has not developed or extremely restricted due to repeated waves, or areas where artificial facilities, grasslands, and observation paths have been constructed. On the other hand, there were 71 lattice points in 14.5% of the total area where the cactus showed 70% or higher dominance. Cacti are randomly distributed in these areas. They have adapted to the microhabitat environment and are found to be opportunistically distributed along the growable locations. Considering that the reproduction of cacti in the habitat is mostly dependent on parthenogenesis, the present distribution seems to reflect the potentially distributable regions of cacti in the habitat. Based on the results of field surveys, a management plan for conservation and protection of the protected areas has been proposed.

A Basic Study on the Establishment of Preservation and Management for Natural Monument(No.374) Pyeongdae-ri Torreya nucifera forest of Jeju (천연기념물 제374호 제주 평대리 비자나무 숲의 보존·관리방향 설정을 위한 기초연구)

  • Lee, Won-Ho;Kim, Dong-Hyun;Kim, Jae-Ung;Oh, Hae-Sung;Choi, Byung-Ki;Lee, Jong-Sung
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.32 no.1
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    • pp.93-106
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    • 2014
  • In this study, Analyze environment of location, investigation into vegetation resources, survey management status and establish to classify the management area for Natural monument No.374 Pyengdae-ri Torreya nucifera forest. The results were as follows: First, Torreya nucifera forest is concerned about influence of development caused by utilization of land changes to agricultural region. Thus, establish to preservation management plan for preservation of prototypical and should be excluded development activity to cause the change of terrain that Gotjawal in the Torreya nucifera forest is factor of base for generating species diversity. Secondly, Torreya nucifera forest summarized as 402 taxa composed 91 familly 263 genus, 353 species, 41 varieties and 8 forms. The distribution of plants for the first grade & second grade appear of endangered plant to Ministry of Environment specify. But, critically endangered in forest by changes in habitat, diseases and illegal overcatching. Therefore, when establishing forest management plan should be considered for put priority on protection. Thirdly, Torreya nucifera representing the upper layer of the vegetation structure. But, old tree oriented management and conservation strategy result in poor age structure. Furthermore, desiccation of forest on artificial management and decline in Torreya nucifera habitat on ecological succession can indicate a problem in forest. Therefore, establish plan such as regulation of population density and sapling tree proliferation for sustainable characteristics of the Torreya nucifera forest. Fourth, Appear to damaged of trails caused by use. Especially, Scoria way occurs a lot of damaged and higher than the share ratio of each section. Therefore, share ratio reduction Plan should be considered through the additional development of tourism routes rather than the replacement of Scoria. Fifth, Representing high preference of the Torreya nucifera forest tourist factor confirmed the plant elements. It is sensitive to usage pressure. And requires continuous monitoring by characteristic of Non-permanent. In addition, need an additional plan such as additional development of tourism elements and active utilizing an element of high preference. Sixth, Strength of protected should be differently accordance with importance. First grade area have to maintenance of plant population and natural habitats. Set the direction of the management. Second grade areas focus on annual regeneration of the forest. Third grade area should be utilized demonstration forest or set to the area for proliferate sapling. Fourth grade areas require the introduced of partial rest system that disturbance are often found in proper vegetation. Fifth grade area appropriate to the service area for promoting tourism by utilizing natural resources in Torreya nucifera forest. Furthermore, installation of a buffer zone in relatively low ratings area and periodic monitoring to the improvement of edge effect that adjacent areas of different class.

Establishment of Pre-Harvest Residue Limit (PHRL) of Fungicides Azoxystrobin and Difenoconazole on Prunus mume fruits (매실 중 살균제 azoxystrobin과 difenoconazole의 생산단계 잔류허용기준 설정)

  • Lee, Dong Yeol;Kim, Yeong Jin;Park, Min Ho;Lee, Seung Hwa;Kim, Sang Gon;Kang, Nam Jun;Kang, Kyu Young
    • The Korean Journal of Pesticide Science
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    • v.17 no.4
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    • pp.307-313
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    • 2013
  • This study was carried out to investigate the residual characteristics of fungicide azoxystrobin and difenoconazole in Prunus mume fruits, and establish pre-harvest residue limits (PHRL) based on dissipation and biological half-lives of fungicide residues. The fungicides were sprayed onto the crop at recommended dosage once and 3 times in 7 days interval, respectively. The samples were harvested at 0, 1, 2, 4, 6, 8, 10, 12 and 14 days after treatment. These residual pesticides were extracted with QuEChERS method, clean-up with $NH_2$ SPE cartridge, and residues were analyzed by HPLC/DAD and GLC/ECD, respectively. Method quantitative limits (MQL) of azoxystrobin were 0.03 mg $kg^{-1}$ and of difenoconazole were 0.006 mg $kg^{-1}$. Average recovery were $93.2{\pm}2.49%$, $85.5{\pm}1.97%$ for azoxystrobin at fortification levels at 0.3 and 1.5 mg $kg^{-1}$, and $100.8{\pm}6.74%$, $87.6{\pm}9.92%$ for difenoconazole at fortification levels at 0.06 and 0.3 mg $kg^{-1}$, respectively. The biological half-lives of azoxystrobin were 5.9 and 5.2 days at recommended dosage once and 3 times in 7 days interval, respectively. The biological half-lives of difenoconazole were 9.3 and 8.0 days at recommended dosage once and 3 times in 7 days interval, respectively. The PHRL of azoxystrobin and difenoconazole were recommended as 5.32 and 1.64 mg $kg^{-1}$ for 10 days before harvest, respectively.

Fulvestrant Does Not Have Antagonistic Effect on 17β-estradiol's Anti-proliferative Action in Cultured Chinese Hamster Ovarian Cell Line (17β-Estradiol의 CHO 세포 항 증식작용에 대한 fulvestrant의 효과)

  • Kim, Hyun Hee;Park, Hyeong Cheol;Min, Gyesik
    • Journal of Life Science
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    • v.24 no.2
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    • pp.173-180
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    • 2014
  • Estrogen can promote or inhibit cellular proliferation depending on tissue cell types and physiological condition and acts through the signal transduction pathways mediated primarily by estrogen receptors. This study examined the effects of fulvestrant (Ful), a well-known antagonist for the estrogen receptor, on the action of $17{\beta}$-estradiol (E2) with respect to the proliferation and apoptosis of Chinese hamster ovarian (CHO) cells. We used different concentrations of E2, Ful, and E2 plus Ful during different treatment durations. Treatment with 15-40 ${\mu}M$ E2 significantly inhibited proliferation in a time-dependent manner, although it had no influence in concentrations up to 1 ${\mu}M$. Interestingly, Ful at 10-40 ${\mu}M$ also inhibited cellular proliferation in both a concentration- and time-dependent manner. In addition, Ful enhanced rather than decreased the inhibitory effect on cellular proliferation by E2 in combined treatment for 10 days. Thus, Ful does not appear to have an antagonistic effect on estrogen's anti-proliferative action in CHO cells. In TUNEL assays to confirm DNA fragmentation by E2 and/or Ful, CHO cells treated with 20 ${\mu}M$ E2 showed a TUNEL-positive reaction in most DAPI-stained nuclei, and cells treated with either 40 ${\mu}M$ Ful or 40 ${\mu}M$ Ful plus 20 ${\mu}M$ E2 also exhibited a TUNEL-positive reaction but at a lower rate compared to the E2-treated cells. These results indicate that Ful does not have an antagonistic effect on estrogen's anti-proliferative action in CHO cells, suggesting that the anti-proliferative and apoptosis-related mechanism(s) through DNA fragmentation by E2 and Ful may be mediated by different signal transduction pathways.

Production of Violacein by a Novel Bacterium, Massilia sp. EP15224 Strain (Violacein을 생산하는 Massilia sp. EP15224 균주)

  • Yoon, Sang-Hong;Baek, Hee-Jin;Kwon, Soon-Wu;Lee, Chang-Muk;Sim, Joon-Soo;Hahn, Bum-Soo;Koo, Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.317-323
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    • 2014
  • Violacein has received much attention due to its various important biological activities, including broad-spectrum antibacterial and antifungal activity, anti-malarial, anti-tumoral, anti-oxidant, and anti-diarrheal activities. EP15224 strain isolated from forest soils in Korea was found to be a new species belonged to the genus Massilia based on its 16S ribosomal DNA sequences. The 16S ribosomal DNA of strain EP15224 displayed 97% homology with Massilia sp. BS-1, the nearest violacein-producing bacterium. Strain EP15224 produced bluish-purple pigment well in a synthetic MM2 medium containing glucose, $(NH_4)_2SO_4$, $Na_2HPO_4{\cdot}7H_2O$, $KH_2PO_4$, $MgSO_4{\cdot}7H_2O$, and 1 mM $\small{L}$-tryptophan. The chemical analysis of the pigment by LC/MS/MS showed that it is violacein with molecular weight of 343.34. This is the second report on the production of violacein by a Massilia species. In this study, the optimal culture conditions for violacein production were established under which 280 mg/l crude violacein was produced : glucose 2 g/l, $(NH_4)_2SO_4$ 1 g/l, $Na_2HPO_4{\cdot}7H_2O$ 2 g/l, $KH_2PO_4$ 1 g/l, $MgSO_4{\cdot}7H_2O$ 0.1 g/l, L-tryptophan 0.24 g/l, 25 ml medium in a 250 ml flask, with an inoculumn size of 10% (v/v), 72 h of cultivation with 250 rpm at $25^{\circ}C$.

The Development of Vulnerable Elements and Assessment of Vulnerability of Maeul-soop Ecosystem in Korea (한국 마을숲 생태계 취약요소 발굴 및 취약성 평가)

  • Lim, Jeong-Cheol;Ryu, Tae-Bok;Ahn, Kyeong-Hwan;Choi, Byoung-Ki
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.34 no.4
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    • pp.57-65
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    • 2016
  • Maeul-soop(Village forest) is a key element of Korean traditional village landscape historically and culturally. However, a number of Maeul-soops have been lost or declined due to various influences since the modern age. For this Maeul-soop that has a variety of conservation values including historical, cultural and ecological ones, attention and efforts for a systematic conservation and restoration of Maeul-soop are needed. The purpose of the present study is to provide information on ecological restoration and sustainable use and management of Maeul-soops based on component plant species, habitat and location characteristics of 499 Maeul-soops spread throughout Korea. Major six categories of threat factors to Maeul-soop ecosystem were identified and the influence of each factor was evaluated. For the evaluation of weight by threat factors for the influence on the vulnerability of Maeul-soop ecosystem, more three-dimensional analysis was conducted using Analytic Hierarchy Process (AHP) analysis method. In the results of evaluation using AHP analysis method, reduction of area, among six categories, was spotted as the biggest threat to existence of Maeul-soops. Next, changes in topography and soil environment were considered as a threat factor of qualitative changes in Maeul-soop ecosystem. Influence of vegetation structure and its qualitative changes on the loss or decline of Masul-soop was evaluated to be lower than that of changes in habitat. Based on weight of each factor, the figures were converted with 100 points being the highest score and the evaluation of vulnerability of Maeul-soop was conducted with the converted figures. In the result of evaluation of vulnerability of Maeul-soops, grade III showed the highest frequency and a normal distribution was formed from low grade to high grade. 38 Maeul-soops were evaluated as grade I which showed high naturality and 10 Maeul-soops were evaluated as grade V as their maintenance was threatened. Also in the results of evaluation of vulnerability of each Maeul-soop, restoration of Maeul-soop's own area was found as top priority to guarantee the sustainability of Maeul-soops. It was confirmed that there was a need to prepare a national level ecological response strategy for each vulnerability factor of Maeul-soop, which was important national ecological resources.

Potential Contamination Sources on Fresh Produce Associated with Food Safety

  • Choi, Jungmin;Lee, Sang In;Rackerby, Bryna;Moppert, Ian;McGorrin, Robert;Ha, Sang-Do;Park, Si Hong
    • Journal of Food Hygiene and Safety
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    • v.34 no.1
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    • pp.1-12
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    • 2019
  • The health benefits associated with consumption of fresh produce have been clearly demonstrated and encouraged by international nutrition and health authorities. However, since fresh produce is usually minimally processed, increased consumption of fresh fruits and vegetables has also led to a simultaneous escalation of foodborne illness cases. According to the report by the World Health Organization (WHO), 1 in 10 people suffer from foodborne diseases and 420,000 die every year globally. In comparison to other processed foods, fresh produce can be easily contaminated by various routes at different points in the supply chain from farm to fork. This review is focused on the identification and characterization of possible sources of foodborne illnesses from chemical, biological, and physical hazards and the applicable methodologies to detect potential contaminants. Agro-chemicals (pesticides, fungicides and herbicides), natural toxins (mycotoxins and plant toxins), and heavy metals (mercury and cadmium) are the main sources of chemical hazards, which can be detected by several methods including chromatography and nano-techniques based on nanostructured materials such as noble metal nanoparticles (NMPs), quantum dots (QDs) and magnetic nanoparticles or nanotube. However, the diversity of chemical structures complicates the establishment of one standard method to differentiate the variety of chemical compounds. In addition, fresh fruits and vegetables contain high nutrient contents and moisture, which promote the growth of unwanted microorganisms including bacterial pathogens (Salmonella, E. coli O157: H7, Shigella, Listeria monocytogenes, and Bacillus cereus) and non-bacterial pathogens (norovirus and parasites). In order to detect specific pathogens in fresh produce, methods based on molecular biology such as PCR and immunology are commonly used. Finally, physical hazards including contamination by glass, metal, and gravel in food can cause serious injuries to customers. In order to decrease physical hazards, vision systems such as X-ray inspection have been adopted to detect physical contaminants in food, while exceptional handling skills by food production employees are required to prevent additional contamination.