• Title/Summary/Keyword: Molecular Characteristics

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Characterization and N-Terminal Amino Acid Sequence Analysis of Catechol 2,3-dioxygenase Isolated from the Aniline Degrading Bacterium, Delftia sp. JK-2 (Aniline 분해세균 Delftia sp. JK-2에서 분리된 catechol 2,3-dioxygenase의 특성 및 N-말단 아미노산 서열분석)

  • 황선영;송승열;오계헌
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.1-7
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    • 2003
  • The aim of this work was to investigate the characterization and sequence of catechol 2,3-dioxygenase isolated from Delfia sp. JK-2, which could utilize aniline as sole carbon, nitrogen and energy source. In initial experiments, several characteristics of C2,3O separated with ammonium sulfate precipitation, DEAE-sepharose were investigated. Specific activity of C2,3O was approximately 4.72 unit/mg. C2,3O demonstrated its enzyme activity to other substrates, catechol and 4-methylcatechol. The optimum temperature of C2,3O was $$Cu^{2+}$^{\circ}C$, and the optimal pH was approximately 8. Metal ions such as $Ag^{+}$, $Hg^{+}$, and $Cu^{2+}$ showed inhibitory effect on the activity of C2,3O. Molecular weight of the enzyme was determined to approximately 35 kDa by SDS-PAGE. N-terminal amino acid sequence of C2,3O was analyzed as $^{1}MGVMRIG-HASLKVMDMDA- AVRHYENV^{26}$, and exhibited high sequence homology with that of C2,30 from Pseudomonas sp. AW-2, Comamonas sp. JS765, Comamonas testosteroni and Burkholderia sp. RPO07. PCR product was amplified with the primers derived from N-terminal amino acid sequence. In this work, we found that the amino acid sequence of Delftia sp. JK-2 showed high sequence homology of C2,3O from Pseudomonas sp. AW-2 (100%) and Comamonas sp. JS765 (97%).

Isolation and Characterization of a Basic Leucine Zipper Gene in Poplar (Populus alba × P. glandulosa) (현사시나무에서 Basic Leucine Zipper 유전자의 분리와 특성 구명)

  • Yoon, Seo-Kyung;Lee, Hyoshin;Bae, Eun-Kyung;Choi, Young-Im;Kim, Joon-Hyeok;Noh, Seol Ah
    • Journal of Korean Society of Forest Science
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    • v.103 no.2
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    • pp.189-195
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    • 2014
  • Basic leucine zipper (bZIP) protein is a regulatory transcription factor that plays crucial roles in growth, development and stress response of plant. In this study, we isolated a PagbZIP1 gene that belonged to Group SE3 of bZIP from Populus alba ${\times}$ P. glandulosa, and investigated its expressional characteristics. The PagbZIP1 is 844 base pairs long and encodes a putative 144-amino-acid protein with an expected molecular mass of 16.6 kDa. The PagbZIP1 has two conserved domains including the basic and leucine zipper portions. Southern blot analysis revealed that two copies of the gene are presented in the poplar genome. PagbZIP1 was specifically expressed in the root and suspension cells. Moreover, the expression of PagbZIP1 was induced by drought, salt, cold and ABA. Therefore, our results indicated that PagbZIP1 might be expressed in response to abiotic stress through the ABA-mediated signaling pathway in poplar.

Characteristics of fruiting bodies color mutants in Pleurotus ostreatus (원형느타리버섯 백색돌연변체의 특성)

  • Lee, Kang-Hyo;Kim, Gyu-Hyun;Kim, Beom-Gi;Yoo, Young-Bok;Sung, Jae-Mo
    • Journal of Mushroom
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    • v.5 no.1
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    • pp.34-38
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    • 2007
  • The white-colored and the dark gray-colored mutants were frequently happened in cultivated areas of Pleurotus ostreatus (Wonhyeong-neutari). These caused conflicts between farmers and spawn companies. Our studies were conducted to elucidate the mechanism of mutagenesis. The results from the studies would provide valuable informations that could be used to prevent the color-related mutation, and also will be applied in breeding programs of P. ostreatus. Oyster mushroom variety, Wonhyeong-neutari, is somatic hybrid of Pleurotus and has genetic makers for arginine, ornithine, proline, riboflavine. Genetic markers analysis of monospore isolates derived from color mutants show identical tendency with that of Wonhyeong-neutari, these results indicate that color mutants were derived from Wonhyeongneutari. Twenty-one and four homokaryons were selected from the white-colored mutant MGL 2205 and gray-colored ASI 2029. All 34 F1 hybrids derived from the white-colored mutant MGL 2205 produce white-color fruiting bodies, indicating that the white color trait is heritable. In the first generation hybrids between the white-colored MGL 2205 and the gray-colored ASI 2029, all 16 hybrids produced pigmented fruiting bodies. Homokaryons isolated from the hybrid MGL 2205 X ASI 2029 were mated with homokaryon tester strains derived from MGL 2205. By these result, we could assumed that white color trait is a heritable character which is controlled by more than one recessive gene.

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Efficient Plant Regeneration from Mature Seed-derived Callus of Italian ryegrass (Lolium multiflorum Lam.) (이탈리안 라이그래스의 성숙종자 유래 캘러스로부터 효율적인 식물체 재분화)

  • Woo, Hyun-Sook;Lee, Sang-Hoon;Lee, Dong-Gi;Kim, Jin-Soo;Won, Sung-Hye;Lee, Byung-Hyun
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.43-48
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    • 2004
  • As an initial step for future genetic manipulations to improve forage characteristics of Italian ryegrass (Lolium multiflorum Lam.), an efficient tissue culture system was established and the factors affecting plant regeneration were evaluated. MS medium containing 5mg/L 2,4-D was optimal for embryogenic callus induction from mature seed and had a strong effect on successive plant regeneration. The plant regeneration frequency was observed at above 70% when embryogenic calli induced were transferred to N6 medium supplemented with 1mg/L 2,4-D and 5mg/L BA. Among several basic media tested, MS and N6 medium were optimal for callus induction and plant regeneration, respectively. Genotype was an important factor in plant regenerability. 'Jeanne' showed the highest regeneration frequency of 73%. A short tissue culture period and high-frequency regeneration system established in this study will be useful for molecular breeding of Italian ryegrass through genetic transformation.

First Report of Fusarium Wilt Caused by Fusarium oxysporum on Kohlrabi in Korea (Fusarium oxysporum에 의한 콜라비 시들음병)

  • Choi, In-Young;Kim, Ju;Ju, Ho-Jong;Park, Ji-Hyun;Shin, Hyeon-Dong
    • Research in Plant Disease
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    • v.21 no.1
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    • pp.27-31
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    • 2015
  • In April 2014, Fusarium wilt was found on kohlrabi seedlings in Iksan, Korea. Symptoms included wilting of foliage, drying and withering of older leaves, and stunting of the plants. The infected plants eventually died during growth. Colonies on potato dextrose agar were pinkish white, and felted with cottony and aerial mycelium. Macroconidia were falcate to almost straight, thin walled and usually 3-septate. Microconidia were usually formed abundantly in false-heads on short monophialides on the hyphae and were hyaline, smooth, oval to ellipsoidal, aseptate or medianly 1-septate, very occasionally 2-septate, slightly constricted at the septa, $4-11{\times}2.5-5{\mu}m$. On the basis of the morphological characteristics and phylogenetic analyses of molecular markers (internal transcribed spacer rDNA and translation elongation factor $1{\alpha}$), the fungus was identified as Fusarium oxysporum. Pathogenicity of a representative isolate was proved by artificial inoculation, fulfilling Koch's postulates. To our knowledge, this is the first report on the occurrence of Fusarium oxysporum on kohlrabi in Korea.

Stem Rot on Ligularia fischeri Caused by Sclerotium rolfsii in Korea (Sclerotium rolfsii에 의한 곰취 흰비단병)

  • Moon, Youn-Gi;Kim, Se-Won;Choi, Jun-Keun;Kwon, Soon-Bae;Shim, Hong-Sik;Ju, Ho-Jong;Choi, In-Young
    • Research in Plant Disease
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    • v.21 no.1
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    • pp.36-39
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    • 2015
  • In June 2012 and 2013, a destructive stem rot symptoms of Ligularia fischeri occurred sporadically in Hoengseong-gun and Pyeongchang-gun Gangwon-do, Korea. The typical symptom included water-soaking on the main stem, rotting, wilting and blighting, which eventually leads to death of the plant. White mycelial mats were spread over lesions and brown sclerotia were formed on stems and near soil surface. The sclerotia were white to brown, spherical or irregular, 1-3 mm in size on potato dextrose agar (PDA), The optimum temperature range of hyphal growth was $25-30^{\circ}C$ and the hyphal diameter was $4-10{\mu}m$. The typical clamp connections were observed in the hyphae of the fungus grown on PDA. The resulting sequence of 695 bp was deposited in GenBank. A BLAST search revealed that sequences of the this isolates showed >99% identity with those of Sclerotium rolfsii. On the basis of the morphological characteristics and phylogenetic analyses of molecular markers ITS rDNA, the fungi were identified as S. rolfsii. A pathogenicity test was carried out to fulfill Koch's postulates. To our knowledge, this is the first report of S. rolfsii on Ligularia fischeri in Korea.

Comparative Analyses for Aroma and Agronomic Traits of Native Rice Cultivars from Central Asia

  • Sarhadi, Wakil Ahmad;Hien, Nguyen Loc;Zanjani, Mehran;Yosofzai, Wahida;Yoshihashi, Tadashi;Hirata, Yutaka
    • Journal of Crop Science and Biotechnology
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    • v.11 no.1
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    • pp.17-22
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    • 2008
  • Aromatic rice has become popular owing to its aroma. Growing demand for aromatic rice has spurred interest in the development of domestic cultivars that offer similar combinations of grain attributes such as texture, cooking characteristics, aroma, and taste. In this study, the most important agronomic attributes and aroma of 26 cultivars from Afghanistan, Iran, and Uzbekistan, and controls from Japan, Thailand, and India were characterized. Also $F_2$ populations derived from the cross between(Jasmine 85 aromatic$\times$Nipponbare non-aromatic) and(Jasmine 85$\times$Basmati 370 aromatic) were obtained. Tasting individual grains, cooking test, 1.7% KOH sensory test, and molecular marker analysis have been applied to distinguish between aromatic and non-aromatic rice. Diversity for some traits of agronomic importance, such as plant height was detected among countries, e.g. Afghan cultivars classified as tall, and Iranian and Uzbek intermediate and short, respectively. Differentiations of panicle, grain, leaf, basal internode, and culm dimension among rice cultivars, indicating the source of rice diversity in Central Asia. According to the results, 6 of 10, 2 of 7, and 0 of 6 of Afghan, Iranian, and Uzbek rice cultivars were scored as aromatic, respectively. Therefore, Afghan cultivars are a good source of aromatic rice germplasm for Central Asia. The expression between aromatic and non-aromatic, and aromatic and aromatic combinations has been evaluated. The observed segregation ratio of these crosses in the $F_2$ populations was tested by $x^2$ analysis against the expected ratio for a single gene. A segregation ratio of 3:1 between non-aromatic and aromatic combination has been detected, while segregation has not been detected between the aromatic and aromatic combinations. Also, parallel results were obtained from the tested aromatic rice cultivars. Thus, our results suggest that a single recessive gene controls aroma in all aromatic rice cultivars.

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Isolation and Characterization of a Novel Polysaccharide Producing Bacillus polymyxa A49 KCTC 4648P

  • Ahn, Sung-Gu;Suh, Hyun-Hyo;Lee, Chang-Ho;Moon, Seong-Hoon;Kim, Hee-Sik;Ahn, Keug-Hyun;Kwon, Gi-Seok;Oh, Hee-Mock;Yoon, Byung-Dae
    • Journal of Microbiology and Biotechnology
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    • v.8 no.2
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    • pp.171-177
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    • 1998
  • The strain A49, which produces a new type of extracellular polysaccharide was isolated from soil samples. From morphological, physiological and biochemical tests, the strain A49 was identified as a Bacillus polymyxa and named Bacillus polymyxa A49. Bacillus polymyxa A49 was found to produce a highly viscous extracellular polysaccharide when grown aerobically in a medium containing glucose as the sole source of carbon. The polysaccharide (A49 POL) showed a homogeneous pattern on gel permeation chromatography (GPC) and its molecular weight was estimated to be about 1.6 mega dalton (mDa). The FT-IR spectrum of A49-POL revealed typical characteristics of polysaccharides. As a result of investigations with HPLC and carbozole assay, A49-POL was found to consist of L-fucose, D-galactose, D-glucose, D-mannose, and D-glucuronic acid, with the molar ratio of these sugars being approximately 1:2:7:50:12. Rheological analysis of A49 POL revealed that it is pseudoplastic and has a higher apparent viscosity at dilute concentrations than does xanthan gum. The consistancy factor of A49 POL was found to be higher, and the flow index of A49 POL lower, than xanthan gum. Its apparent viscosity was comparatively unstable at various temperatures. the A49 POL showed the highest apparent viscosity at pH 3. When salts were added to A49 POL solution, the solution was compatible with up to 10% KCl, 35% NaCl, 55% $CaCl_2$, 55% $MgCl_2$, 55% $K_2HPO_4$, and 110% $Ca({NO_3})_2$, respectively.

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Isolation and Characterization of Three Kinds of Lipopeptides Produced by Bacillus subtilis JKK238 from Jeot-Kal of Korean Traditional Fermented Fishes (한국 전통젓갈에서 분리한 Bacillus subtilis JKK238 균주 유래 세 종류 Lipopeptide의 분리 및 특성)

  • Yoon Sang-Hong;Kim Jung-Bong;Lim Yoong-Ho;Hong Seong-Ryeul;Song Jae-Kyeung;Kim Sam-Sun;Kwon Soon-Wo;Park In-Cheol;Kim Soo-Jin;Yeo Yun-Soo;Koo Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.295-301
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    • 2005
  • About seven hundred bacterial strains were collected from Jeot-Kal, a Korean traditional fermented fishes, in various Korean districts. One of the strains designated JKK238 has its ability to antagonize in vitro the growth of a wide variety of plant pathogenic fungi responsible for diseases of economical importance. The JKK238 strain was isolated from Oh-Jeot, a kind of fermented shrimps, of Kangkyeung in Korea, and was identified as Bacillus subtilis based on its physiological characteristics, fatty acids compositions of cellular wall, and 16S rDNA sequence analysis. We isolated simply antimicrobial lipopeptides (AMLP) by $25\%$ ammonium sulfate precipitation of 3 days-old tryptic soy broth cultures of the JKK238 strain. Further analysis of AMLP revealed that B. subtilis JKK238 produces a wide variety of antifungal lipopeptide isomers from the iturin, fengycin and surfactin families simultaneously. Above results indicate that the JKK238 strain can be added to the limited number B. subtilis strains reported to co-produce the three kinds of lipopeptide families.

Selection and Characteristics of Bacteriocin-Producing Microorganism to Utilize in Anti-Bacterial Rice Brain Protein Film Production (항균성 미강 단백질 필름 개발을 위한 Bacteriocin 생성균주의 선별 및 특성)

  • Kim, Eun-Joung;Kim, Kyung-Mi;Han, Hye-Kyung;Kim, Young-Ho;Kwon, Ki-Sung;Bae, Dong-Ho
    • Applied Biological Chemistry
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    • v.46 no.4
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    • pp.285-290
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    • 2003
  • This study was conducted to select the bacteriocin-producing microoreanism cultivated in the rice bran culture and to characterize the produced bacteriocin for the further purpose of economical and anti-bacterial rice bran protein film. Pseudomonas putida 21025 was cultivated from rice bran and identified as a producer of a bacteriocin which showed bactericidal activity against Pseudomonas aeruginosa 9027. Bacteriocin produced by Pseudomonas putida 21025 showed a broad spectrum of activity against spoilage and soil bacteria. The activity of the bacteriocin produced by Pseudomonas putida 21025 decreased after 1 hr of staying at the temperature of $50^{\circ}C$, and with the presence of some organic solvents, except hexane and ethanol. However, the bacteriocin activity was stable throughout the pH ranges of 6-9 for 2 hrs, at the temperature lower than $50^{\circ}C$, and with the presence of ethanol for 3 hrs. The bacteriocin was partially purified by 50% ammonium sulfate precipitation followed by subsequent dialysis. Direct detection of the partially purified bacteriocin on SDS-PAGE suggested that it had an apparent molecular mass of about 21.6 kDa.