• Title/Summary/Keyword: rhizobium

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Transformation of Medicago truncatula with rip1-GUS Gene

  • Nam Young-Woo;Song Dae-Hae
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.5
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    • pp.434-439
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    • 2004
  • Medicago truncatula is a model plant for molecular genetic studies of legumes and plant-microbe interactions. To accelerate finding of genes that play roles in the early stages of nodulation and stress responses, a trans-genic plant was developed that contains a promoter­reporter fusion. The promoter of rip], a Rhizobium-induced peroxidase gene, was fused to the coding region of $\beta-glucuronidase (GUS)$ gene and inserted into a modified plant transformation vector, pSLJ525YN, in which the bar gene was preserved from the original plasmid but the neomycin phosphotransferase gene was replaced by a polylinker. Transformation of M. truncatula was carried out by vacuum infiltration of young seedlings with Agrobacterium. Despite low survival rates of infiltrated seedlings, three independent transformants were obtained from repeated experiments. Southern blot analyses revealed that 7 of 8 transgenic plants of the T 1 generation contained the bar gene whereas 6 $T_1$ plants contained the GUS gene. These results indicate that vacuum infiltration is an effective method for transformation of M. truncatula. The progeny seeds of the transgenic plants will be useful for mutagenesis and identification of genes that are placed upstream and may influence the expression of rip] in cellular signaling processes including nodulation.

Approach Toward the Creation of Nonnarcotic Opium Poppy - Morphological and Genetical Analysis on 'Thebaine Poppy' -

  • Kawano, Noriaki;Yoshimatsu, Kayo;Kiuchi, Fumiyuki
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.361-366
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    • 2005
  • Opium poppy (Papaver somniferum L.) is one of the most important medicinal plants, which is used as a sole commercial source of narcotic analgesic, morphine. The transformant of opium poppy we have established by infection of Rhizobium rhizogenes (formerly Agrobacterium rhizogenes) strain MAFF03-01724 showed aberrant morphology and altered opium alkaloid composition. The major alkaloid produced by this transformant was thebaine (16.3%, opium dry weight) instead of morphine. It is likely that this 'thebaine poppy' phenotype was caused by the integration of T-DNA(s) into the poppy genome DNA, and their inserted loci are of great interest. To gain an insight into the mechanism of nonnarcotic thebaine accumulation for the further approach toward the creation of 'codeine poppy' which produces codeine as a major alkaloid, the genetical and morphological analyses on the transformant was carried out. Here we report the results of the detailed analysis on the T-DNA inserted loci of T0 transfromant and the correlation between opium alkaloid composition and segregated T-DNA integration pattern in the self-pollinated T1 transformants.

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Diversity of Nitrifying and Denitrifying Bacteria in SMMIAR Process (완전침지형 회전매체공정 내 질산화 및 탈질 관련 미생물의 군집 분포)

  • Quan, Zhe-Xue;Lim, Bong-Su;Kang, Ho;Yoon, Kyung-Yo;Yoon, Yeo-Gyo
    • Journal of Korean Society on Water Environment
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    • v.22 no.6
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    • pp.1014-1021
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    • 2006
  • SMMIAR (Submerged Moving Media Intermittent Aeration Reactor) Process is a very efficient system which remove ammonia to nitrogen gas in one reactor. In this study, we determined the diversity of ammonia oxidizing bacteria and denitrifying bacteria using specific PCR amplification and the clone library construction. An ammonia monooxygenase gene(amoA) was analyzed to investigate the diversity of nitrifiers. Most of amoA gene fragments (27/29, 93%) were same types and they are very similar (>99%) to the sequences of Nitrosomonas europaea and other clones isolated from anoxic ammonia oxidizing reactors. ANAMMOX related bacteria have not determined by specific PCR amplification. A nitrite reductase gene(nirK) was analyzed to investigate the diversity of denitrifying bacteria. About half (9/20, 45%) of denitrifiers were clustered with Rhodobacter and most of others were clustered with Mesorhizobium (6/20, 30%) and Rhizobium (3/20, 15%). All of these nirK gene clones were clustered in alpha-Proteobacteria and this result is coincide with other system which also operate nitrification and denitrification in one reactor. The molecular monitoring of the population of nitrifiers and denitrifiers would be helpful for the system stabilization and scale-up.

Characterization and biovar. cetermination of agrobacterium tumefaciens T7 isolated in Korea (한국에서 분리한 agrobacterium tumefaciens T7의 특성과 biovar.결정)

  • Rhee, Y.;Kim, C. J.;Kim, S. H.;Yoo, I. D.;Mheen, T. I.
    • Korean Journal of Microbiology
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    • v.25 no.1
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    • pp.17-22
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    • 1987
  • For the purpose of securing of strains which can be usefully utilized to study symbiosis between Rhizobium and legume plant, A. tumefaciens T7 was isolated and characterized and then subgroup biovar was determined. A. tumefaciens T7 induced smooth tumor like nopaline type one and did not grow at $37^{\circ}C$ and in the presence of 2% NaCl on yeast extract mannitol medium. The strain was able to grow on the New and Kerr selective media and utilize erythritol but not phenylalanine, tryptophan, and tartarate as a sole carbon source. Negative results were obtained from 3-keto-lactose production and oxidase test. The strain produced alkalifrom malonate and citrate and showed acid litmus milk reaction At least two large plasmids were detected in the cell lysate. According to all of these results, it could be concluded that subdivision of isolated strain was biovar 2.

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A report of 22 unrecorded bacterial species in Korea, isolated from the North Han River basin in 2017

  • Joung, Yochan;Park, Miri;Jang, Hye-Jin;Jung, Ilsuk;Cho, Jang-Cheon
    • Journal of Species Research
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    • v.7 no.3
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    • pp.193-201
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    • 2018
  • Culturable bacterial diversity was investigated using freshwater and sediment samples collected from the North Han River basin in 2017, as a part of the research program 'Survey of freshwater organisms and specimen collection'. Over a thousand bacterial strains were isolated from the samples and identified based on 16S rRNA gene sequences. Among the bacterial isolates, 22 strains showing higher than 98.7% sequence similarity with validly published bacterial species, but not reported in Korea, were classified as unrecorded species in Korea. The 22 bacterial strains were phylogenetically diverse and assigned to 6 classes, 11 orders, 15 families, and 21 different genera. At the generic level, the unreported species were affiliated with Flavobacterium of the class Flavobacteria, Flexibacter of the class Cytophagia, Blastomonas, Brevundimonas, Elstera, Rhizobium, Roseomonas, Sphingomonas, and Xanthobacter of the class Alphaproteobacteria, Albidiferax, Cupriavidus, Curvibacter, Ferribacterium, Hydrogenophaga, Iodobacter, Limnohabitans, Polaromonas, Undibacterium, and Variovorax of the class Betaproteobacteria, Pseudomonas of the class Gammaproteobacteria, and Arcobacter of the class Epsilonproteobacteria. The unreported bacterial species were further characterized by examining Gram reaction, colonial and cellular morphology, and biochemical properties. The detailed descriptions of 22 strains of the unreported bacterial species are also provided.

A report of 37 unrecorded anaerobic bacterial species isolated from the Geum River in South Korea

  • Lee, Changsu;Kim, Joon Yong;Kim, Yeon Bee;Kim, Juseok;Ahn, Seung Woo;Song, Hye Seon;Roh, Seong Woon
    • Journal of Species Research
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    • v.9 no.2
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    • pp.105-116
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    • 2020
  • A total of 37 anaerobic bacteria strains within the classes Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Bacteroidia, Flavobacteriia, Bacilli, Clostridia, and Fusobacteriia were isolated from freshwater and sediment of the Geum River in Korea. The unreported species were related with Rhizobium and Oleomonas of the class Alphaproteobacteria; Acidovorax, Pseudogulbenkiania, and Aromatoleum of the class Betaproteobacteria; Tolumonas, Aeromonas, Cronobacter, Lonsdalea, and Phytobacter of the class Gammaproteobacteria; Bacteroides, Dysgonomonas, Macellibacteroides, and Parabacteroides of the class Bacteroidia; Flavobacterium of the class Flavobacteriia; Bacillus and Paenibacillus of the class Bacilli; Clostridium, Clostridioides, Paraclostridium, Romboutsia, Sporacetigenium, and Terrisporobacter of the class Clostridia; and Cetobacterium and Ilyobacter of the class Fusobacteriia. A total of 37 strains, with >98.7% 16S rRNA gene sequence similarity with validly published bacterial species, but not reported in Korea, were determined to be unrecorded anaerobic bacterial species in Korea.

Molecular Genetics of the Model Legume Medicago truncatula

  • Nam, Young-Woo
    • The Plant Pathology Journal
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    • v.17 no.2
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    • pp.67-70
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    • 2001
  • Medicago truncatula is a diploid legume plant related to the forage crop alfalfa. Recently, it has been chosen as a model species for genomic studies due to its small genome, self-fertility, short generation time, and high transformation efficiency. M. truncatula engages in symbiosis with nitrogen-fixing soil bacterium Rhizobium meliloti. M. truncatula mutants that are defective in nodulation and developmental processes have been generated. Some of these mutants exhibited altered phenotypes in symbiotic responses such as root hair deformation, expression of nodulin genes, and calcium spiking. Thus, the genes controlling these traits are likely to encode functions that are required for Nod-factor signal transduction pathways. To facilitate genome analysis and map-based cloning of symbiotic genes, a bacterial artificial chromosome library was constructed. An efficient polymerase chain reaction-based screening of the library was devised to fasten physical mapping of specific genomic regions. As a genomics approach, comparative mapping revealed high levels of macro- and microsynteny between M. truncatula and other legume genomes. Expressed sequence tags and microarray profiles reflecting the genetic and biochemical events associated with the development and environmental interactions of M. truncatula are assembled in the databases. Together, these genomics programs will help enrich our understanding of the legume biology.

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Effects of $NO_3^-$ Gradients on Nitrogen Fixation, Nitrate Reduction and Ureide Content of Soybean (대두의 공소개정, 공산환원 및 Ureide함량에 미치는$NO_3^-$의 영향)

  • 추연식
    • Journal of Plant Biology
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    • v.30 no.3
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    • pp.205-213
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    • 1987
  • Soybean, inoculated with effective Rhizobium japonicum 110, were grown by sand culture with nutrient solution containing either of 0, 1, 3, 10 or 30mM NO3-/l, and analyzed growth characteristics, NR activity, N2-fixation activity, and changes of ureide contents during the growing period. The amount of nodule formation decreased abruptly by nitrate treatment, the maximum nodule dry weight was 1.59, 1.05, 0.78, 0.09 and 0.008 g plant-1, respectively for each treatment on the 98th day. Specfic activity of N2-fixation showed the maximum rates of 140, 101, 37, 5 and 2.2 nM dw.mg-1.hr-1, respectively for each treatment in the earlier growth period. The maximum acetylene reduction activity on the 98th day after sowing was 81.5, 35.3, 14.3, 0.1 and 0.0045 $\mu$M C2H4 plant-1.hr-1, respectively for 0, 1, 3, 10 and 30 mM of NO3- gradients. Nitrate reduction activity increased along with nitrate gradients, and decreased abruptly with age. Relative abundance of ureides in plant organs was high in reproductive growth, and showed the maximum value in fully symbiotic dependent plant. Relative abundance of ureides in stem is a useful indication for the evaluation of nitrogen fixation in nodules of symbiotic plant.

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A report of 31 unrecorded bacterial species isolated from freshwater

  • Hyangmi Kim;Sanghwa Park;Kyung June Yim;Ja Young Cho;Eui-Jin Kim
    • Korean Journal of Environmental Biology
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    • v.40 no.4
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    • pp.442-454
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    • 2022
  • A total of 31 bacterial strains were isolated from the Geum River basin in the Republic of Korea during our investigation of indigenous prokaryotic species. The isolated bacterial strains had high 16S rRNA gene sequence similarity (>98.7%) with those of validly published bacterial species, which have not been reported in Republic of Korea. The 31 bacterial strains were phylogenetically diverse and assigned to 4 phyla, 8 classes, 18 orders, 21 families, and 27 genera. At the genus level, the unreported species were affiliated with Kineococcus, Pedococcus, Rhodoluna, Salinibacterium, Rhodoluna, Arthrobacter, Williamsia, Nakamurella, Nocardioides of the class Actinobacteria, Patulibacter of the class Thermoleophilia, Pontibacter, Hymenobacter of the class Cytophagia, Flavobacterium of the class Flavobacteriia, Geomicrobium of the class Bacilli, Brevundimonas, Gellertiella, Rhizobium, Paracoccus, Taonella, Sphingomonas of the class Alphaproteobacteria, Burkholderia, Polaromonas, Hydrogenophaga, Chitinilyticum, Azospira, Zoogloea of the class Betaproteobacteria, and Pseudomonas of the class Gammaproteobacteria. The unreported bacterial species were further characterized by examining their morphological, cultural, physiological, and biochemical properties. The detailed descriptions of the 31 bacterial strains were provided.

A report of 12 unrecorded bacterial species isolated from Suncheon Bay in Korea

  • Seok Won Jang;Jung Hye Eom;Sanghwa Park
    • Korean Journal of Environmental Biology
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    • v.40 no.4
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    • pp.405-412
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    • 2022
  • Suncheon Bay Ecological Park, possessing abundant fisheries and biological diversity, was registered as a Ramsar wetland in Korea. Approximately 300 bacterial strains were isolated from the Suncheon Bay in a comprehensive study of indigenous prokaryotic species conducted during 2019-2020 in South Korea. A total of 12 bacterial strains were identified using 16S rRNA gene sequencing, demonstrating >98.7% sequence similarity with validly published species. These species were determined to be unrecorded bacterial species in Korea. A total of six strains were isolated from brackish water and Phragmites communis Trin (reed) species. These unrecorded species were phylogenetically diverse and belonged to three classes, six orders, and ten genera. Regarding the genus and class levels, the previously unrecorded species belonged to Jiella, Martelella, Rhizobium, Paracoccus, Rhodovulum, and Altererythrobacter of the class Alphaproteobacteria; Mycolicibacterium, Demequina, and Microbacterium of the class Actinobacteria; Confluentibacter of the class Flavobacteria. The twelve species were further characterized by gram staining, colony and cell morphology, biochemical properties, and phylogenetic position.