• Title/Summary/Keyword: plant cultured cells

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Antinociceptive, anti-inflammatory, and cytotoxic properties of Origanum vulgare essential oil, rich with β-caryophyllene and β-caryophyllene oxide

  • Moghrovyan, Armenuhi;Parseghyan, Lilya;Sevoyan, Gohar;Darbinyan, Anna;Sahakyan, Naira;Gaboyan, Monica;Karabekian, Zaruhi;Voskanyan, Armen
    • The Korean Journal of Pain
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    • v.35 no.2
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    • pp.140-151
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    • 2022
  • Background: Essential oils are of great interest for their analgesic and anti-inflammatory properties. We aimed to study the content of the essential oil of the Origanum vulgare of the Armenian highlands (OVA) in different periods of vegetation and to investigate its antinociceptive and anti-inflammatory effects in mice (in vivo) and cytotoxic action in cultured cells (in vitro). OVA essential oil was extracted from fresh plant material by hydro-distillation. Methods: For OVA essential oil contents determination the gas chromatography-mass spectrometry method was used. Formalin and hot plate tests and analysis of cell viability using the methyl-thiazolyl-tetrazolium (MTT) assay were used. Results: The maximal content of β-caryophyllene and β-caryophyllene oxide in OVA essential oil was revealed in the period of blossoming (8.18% and 13.36%, correspondently). In the formalin test, 4% OVA essential oil solution (3.5 mg/mouse) exerts significant antinociceptive and anti-inflammatory effects (P = 0.003). MTT assay shows approximately 60% cytotoxicity in HeLa and Vero cells for 2.0 µL/mL OVA essential oil in media. Conclusions: The wild oregano herb of Armenian highlands, harvested in the blossoming period, may be considered as a valuable source for developing pain-relieving preparations.

Differential Expression of a Chimeric nos-npt II Gene in 9 Years Old Hybrid Poplars (Populus koreana x P. nigra)

  • Noh, Eun Woon;Lee, Jae Soon;Choi, Young Im;Lee, Hyo Shin;Bae, Eun Kyung;Lee, Ji Hee
    • Journal of Plant Biotechnology
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    • v.6 no.1
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    • pp.15-19
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    • 2004
  • The expression of a chimeric transgene (nos-npt II) has been examined in 9 years old transgenic poplars (Populus koreana x P. nigra) growing in a nursery. The expression of the gene in twenty six independentely transformed plants were examined by 1) enzyme (NPT II) assay, 2) RT-PCR, and 3) resistance to kanamycin. High NPT II activities in young leaves of all the transformed plants were found even without a selection pressure for antibiotics for 9 years. However, the activity varied with the positions of leaves in the stem in that young leaves showed higher activity than did mature tissues. When leaf segments were cultured in the presence of 150 mg/l kanamycin, only those from young leaves produced vigorously growing callus. However, as in the case of NPTII assay, the leaf segments from mature leaves did not form callus well on the media. RT-PCR with nptII specific primers also showed that amplification products were observed only when RNAs from young tissues were used. The total RNA gel showed that while RNA in young leaves are relatively stable and in a large quantity, those in old leaves were mostly degraded. All the above results suggest that the gene is transcriptionally active only in young tissue even though it is attached to a constituitive promoter. Therefore, the expression of foreign gene in poplar plants seemed to be affected by the metabolic state of the cells and thus vary greatly with the developmental stages and the age of tissue.

Morphogenetic Alterations of Alternaria alternata Exposed to Dicarboximide Fungicide, Iprodione

  • Kim, Eunji;Lee, Hye Min;Kim, Young Ho
    • The Plant Pathology Journal
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    • v.33 no.1
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    • pp.95-100
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    • 2017
  • Fungicide-resistant Alternaria alternata impede the practical control of the Alternaria diseases in crop fields. This study aimed to investigate cytological fungicide resistance mechanisms of A. alternata against dicarboximide fungicide iprodione. A. alternata isolated from cactus brown spot was cultured on potato-dextrose agar (PDA) with or without iprodione, and the fungal cultures with different growth characteristics from no, initial and full growth were observed by light and electron microscopy. Mycelia began to grow from one day after incubation (DAI) and continued to be in full growth (control-growth, Con-G) on PDA without fungicide, while on PDA with iprodione, no fungal growth (iprodione-no growth, Ipr-N) occurred for the first 3 DAI, but once the initial growth (iprodione-initial growth, Ipr-I) began at 4-5 DAI, the colonies grew and expanded continuously to be in full growth (iprodione-growth, Ipr-G), suggesting Ipr-I may be a turning moment of the morphogenetic changes resisting fungicidal toxicity. Con-G formed multicellular conidia with cell walls and septa and intact dense cytoplasm. In Ipr-N, fungal sporulation was inhibited by forming mostly undeveloped unicellular conidia with degraded and necrotic cytoplasm. However, in Ipr-I, conspicuous cellular changes occurred during sporulation by forming multicellular conidia with double layered (thickened) cell walls and accumulation of proliferated lipid bodies in the conidial cytoplasm, which may inhibit the penetration of the fungicide into conidial cells, reducing fungicide-associated toxicity, and may be utilized as energy and nutritional sources, respectively, for the further fungal growth to form mature colonies as in Ipr-G that formed multicellular conidia with cell walls and intact cytoplasm with lipid bodies as in Con-G.

Resistance of Plants to Herbicide (제초제(除草劑)에 대한 식물(植物)의 저항성(抵抗性))

  • Kim, Kil-Ung
    • Korean Journal of Weed Science
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    • v.4 no.1
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    • pp.96-106
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    • 1984
  • Changes in weed floras and development of plant resistance to herbicides seemed to be closely related with increased and repeated use of herbicides. Herbicide use increased from 5% of the total consumption of pesticide in 1950 to 45% in 1976 in world basis. About 200 herbicides have been introduced to agriculture so as to control about 206 weed species which have been recorded important to human beings. In Korea, there was about 351 times in increased use of herbicides from 1966 to 1982. Interspecific selection by herbicide is mainly responsible for changes in weed floras and resulted in varying tolerance or susceptibility to herbicides, together with the changes of agricultural practices. The present trend toward continuous cereal cultivation throughout world will lead to type of changes in weed floras favorable to therophyte which can survive under unfavorable conditions as seeds rather than the types of geophyte which can survive unfavorable seasons as buds placed below soil surface. However, geophyte such as Sagitaria pygmaea, and Scirpus jurtcoides, and Cyperus rotundus and Cynodon dactylon in temperate warm climate become severe paddy weeds, presumably because of the removal of annual weeds by herbicides. Since differential tolerance to 2,4-D was firstly reported in Agrostis stolofera, about 30 species of weeds in 18 genera are presently known to have developed resistance to triazine herbicides. Resistance of weed biotypes to triazine herbicide is not mainly due to limited absorption and translocation or to the difference in metabolism, but is the result of biochemical changes at the site of metabolic activity, such as a loss of herbicide affinity for triazine binding site in the photosystem II complex of the chloroplast membrane. Genetical study showed that plastid resistance to triazine was wholly inherited through cytoplasmic DNA in the case of Brassica campestris. Plant tissue culture method can be utilized as an alternate mean of herbicide screening and development of resistance variants to herbicides as suggested by Chaleff and Parsons. In this purpose, one should be certain that the primary target process is operational in cell culture. Further, there are a variety of obstacles in doing this type of research, particularly development of resistance source and it's regeneration because cultured cells and whole plants represent different developmental state.

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Occurrence of Leptosphaerulina Leaf Blight on Kentucky Bluegrass Caused by Leptosphaerulina trifolii (Leptosphaerulina trifolii에 의한 Kentucky Bluegrass의 Leptosphaerulina 잎마름병 발생)

  • Kim, Jeong-Ho;Shim, Gyu-Yul;Kim, Young-Ho
    • Research in Plant Disease
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    • v.16 no.1
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    • pp.94-96
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    • 2010
  • In May of 2004 through 2007, Leptosphaerulina leaf blight caused by Leptosphaerulina trifolii occurred on Kentucky bluegrass (Poa pratensis) at golf courses in Gangwon Province, Korea. Symptoms on the turfgrass caused by L. trifolii were leaf blights, dying from the leaf tip downwards to the crown, which appeared patches in the field because of local pockets of severely infected (blighted) grass. Perithecia were produced on old or weak leaves, including club-shaped asci, each of which contained 8 pale brown muriform ascospores with cross and longitudinal septa. Ascospores of the fungus isolated from the diseased leaf tissue and cultured on potato-dextrose agar (PDA) were muriform multicellular (composed of 3-6 cells) and $23.4-40.5{\times}7.8-15.6{\mu}m$ in size with 3-4 transverse and 0-3 longitudinal septa, which were morphologically identical to L. trifolii reported previously. DNA sequences of ribosomal RNA gene (internal transcribed spacer) of the fungus were homologous with similarity of 99% to those of L. trifolii isolates in GenBank database, confirming the identity of the causal agent of the disease. Pathogenicity of the fungus was also confirmed on the creeping bentgrass by Koch's postulates. This is first report of Leptosphaerulina leaf blight on turfgrass caused by L. trifolii in Korea.

Active Transport of Acidic Amino Acids in Suspension Cultured Brassica sp. Cells (배추과 식물현탁배양 세포내에서 산성 아미노산의 능동수송)

  • 조봉희
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.137-142
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    • 1995
  • The acidic amino acids, aspartate and glutamate, which have a negative charge in physiological pH, possess the same transport system as neutral amino acids according to the competitive inhibitory studies with the neutral amino acids. The neutral amino acids cotransported with one H+ per molecule, and one K+efflux per one molecule for charge compensation (Cho,1994), but the acidic amino acids cotransported with two H+ per one molecule, and one K+ efflux per one molecule. The active transport system, which possess the same carrier but cotransported with the different number of H+, reported for the first time. from the results, we can see that one of cotransported H+ protonated at first carboxyl group of pK$_3$ of acidic amino acids, and then as a neutral form cotransported with H+ Therefore, Brassica possess two amino acids transport system for 20 amino acids, namely general - and basic amino acids transport system. The evolutionary meaning of amino acid carriers described with other reported plants.

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Cryopreservation of Embryogenic Callus in Sweetpotato cv. 'Yulmi' (고구마품종 '율미' 배발생 캘러스의 초저온 동결보존)

  • Park, Jong-Suk;Kim, Suk-Weon;In, Dong-Su;Eun, Jong-Seon
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.109-113
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    • 2003
  • Cryopreservation of embryogenic callus derived from apical meristem culture was attempted by slow prefreezing method (two-step method) with various cryoprotectants in sweetpotato cv. 'Yulmi' Precultured embryogenic calli on medium containing 10 mg/L ABA prior to slow prefreezing in liquid nitrogen indicated higher survival rate than 1.0 mg/L ABA preteatment. The cryoprotectant comprising 1.28 M DMSO in 0.4 M sucrose solution gave the best survival (over 46%) of sweetpotato cells exposed to liquid nitrogen as determined by TTC reduction and FDA staining method. Cryopreserved calli cultured on MS medium with 1.0 mg/L 2,4-D were grown for 4 weeks in the dark and induced embryos after another 4 weeks. They were subcultured on MS medium supplemented with 0.1 mg/L 2,4-D+0.1 mg/L kinetin for 2 weeks and regenerated into normal plantlets in MS basal medium.

Effect of Low Temperature Pretreatment on Pollen Dimorphism and Embryo Formation in Anther Culture of Platycodon grandiflorum (도라지 (Platycodon grandiflorum) 약배양에서 저온처리가 화분 2형현상 및 배형성에 미치는 영향)

  • 고정애
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.149-156
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    • 1999
  • In order to investigate the effect of low temperature pretreatment on pollen dimorphism and embryo formation in anther culture of Platycodon grandiflorum, the anthers with microspore at the uninucleate stage were cultured on Murashige and Skoog medium supplemented with 0.5mg/L NAA and 1.0 mg/L BA. The low temperature pretreatment have clear effect on the frequencies of S pollen grains, symmetrical binucleate microspores (B type of S pollen), multinucleate and multicelled pollen grains. Especially, after low temperature pretreatment at 8$^{\circ}C$ for 5 days increased the frequency of S pollen grain (20.6%) in vivo. In addition, the highest frequency of callus induction (54.9%) and embryo formation (9.9%) were obtained from the anther pretreatment at 8$^{\circ}C$ or 5 days. Three distinct pathways could be recognized in the androgenesis, one involving mainly the vegetative cell, the second starting with the vegetative and the generative cell, respectively, and the third accompaning with two equal vegetative type cells in the pollen grains.

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Effect of Abscisic Acid on the Number of Somatic Embryo Cotyledons in Tissue Cultures of Aralia cordata Thunb. (땅두릅(Aralia cordata Thunb.)의 조직배양에서 체세포배의 자엽 수 변화에 미치는 ABA의 영향)

  • 이강섭;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.5
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    • pp.287-291
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    • 1994
  • In order to elucidate the effect of abscisic acid (ABA) on the abnormality of somatic embryos, somatic embryos were induced from embryogenic cell clumps derived from cotyledon segment of Aralia cordata. When embryogenic cell clumps were pretreated medium containing 0.2 mg/L ABA for 3 weeks before transferring to MS basal medium, the frequency of embryos with normal cotyledons enhanced 68% as compared with control. However when clumps pretreated in medium containing 0.2 mg/L ABA were transferred to medium containing 0.1 mg/L ABA, the Sequency decreased to about 29%. In the case of globular embryos cultures in medium containing various concentrations of ABA (0.01 to 1.0mg/L), the frequency of dicotyledonary embryo formation decreased propotionally to ABA concentration. Also, when somatic embryos at various stages were cultured in medium containing ABA, those with polycotyledons appeared at higher frequency.

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Outbreak of Anguillid herpesvirus-1 (AngHV-1) infection in cultured shortfin eel (Anguilla bicolor) in Korea (양식 동남아산 뱀장어, Anguilla bicolor의 Anguillid herpesvirus-1 (AngHV-1) 감염증)

  • Park, Sung-Woo;Jung, Eun-Bin;Kim, Dong-Wan
    • Journal of fish pathology
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    • v.25 no.3
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    • pp.151-158
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    • 2012
  • Diseased eel (Anguilla bicolor) displayed severe hemorrhages in the gills, and congestion and swelling in the liver. During the epizootic, the water temperature was $28^{\circ}C$ and the morality rates were about 5%. No parasites were found on the gills and skin. Bacteria were not cultured from any internal organs using TSA or SS agar at $28^{\circ}C$ for 48 hrs. Histopathologically, the gills showed epithelial hyperplasia in the base of secondary gill lamellae and hemorrhages in the capillaries. Some cells in the proliferated interlamellar epithelia exhibited marginal hyperchromatosis. And severe vacuolated changes in the parenchymal cells and congestion in the central veins were observed in the liver. The specific amplicon (396 bp) was detected from gills and opercula of affected eel PCR using Anguillid herpesvirus-1 (AngHV-1) -specific primer sets HVAPOLVPSD (5-'GTG TCG GGC TTT GTG GTG C-3') and HVAPOLOOSN (5'-CAT GCC GGG AGT CTT TTT GAT-3'). Sequencing analysis of the amplicon demonstrated that this gene was 99% homologous to the AngHV-1 sequence deposited in GenBank. This is the first report of AngHV-1 outbreak in the farmed shortfin eels (A. bicolor) in Korea. When diseased fish were maintained for 10 days at water temperatures of $32^{\circ}C$ and $35^{\circ}C$, the cumulative mortalities were 100% and 10%, respectively. Even though the AngHV-1 genome in the gills from the eel kept at $35^{\circ}C$ was detected using PCR, the structure of gill filaments was similar with that of normal fish. Increasing the water temperature to $35^{\circ}C$ was an effective way to diminish the mortality of AngHV-1 affected eel.