• 제목/요약/키워드: 45S and 5S rDNA

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Change of Microbial Communities in Kimchi Fermentation at Low Temperature (김치의 저온 발효 중 미생물 변화 양상)

  • 박정아;허건영;이정숙;오윤정;김보연;민태익;김치경;안종석
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.45-50
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    • 2003
  • The diversity and change of microbial communities during kimchi fermentation at $4^{\circ}C$ were analyzed by denaturing gradient gel electrophoresis (DGGE). Kimchi samples were taken every 5 days over the fermentation periods (for 60 days) to extract total DNA for DGGE analysis. Touchdown polymerase chain reaction was performed to amplify the V3 region of 16S rRNA gene. Sequencing results of partial 16S rDNA amplicons from DGGE profiles revealed that lactic acid bacteria (LAB), especially Weissella koreensis, Lactobacillus sakei and Leuconostoc gelidum were dominants in kimchi fermentation at $4^{\circ}C$. And we knew that W. koreensis steadily existed throughout the whole fermentation period, also Lb. sakei and Leuc. gelidum appeared from 10th day and 30th day of fermentation time, respectively and then these species were to be dominant microorganisms.

Centromere Repeat DNA Originated from Brassica rapa is Detected in the Centromere Region of Raphanus sativus Chromosomes

  • Hwang, Yoon-Jung;Yu, Hee-Ju;Mun, Jeong-Hwan;Bok, Kwang;Park, Beom-Seok;Lim, Ki-Byung
    • Horticultural Science & Technology
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    • v.30 no.6
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    • pp.751-756
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    • 2012
  • Fluorescence in situ hybridization (FISH) is a powerful tool for the detection of DNA sequences in the specific region of the chromosomes. As well as for the integrated physical mapping, FISH karyotype analysis has to be preceded. Karyotype of Raphanus sativus 'Wonkyo 10039' was analyzed by a dual-color FISH technique; using various repetitive DNA probes, including 5S rDNA, 45S rDNA, and centromere retrotransposon. The length of the somatic metaphase chromosome ranged from 1.35 to $2.06{\mu}m$ with a total length of $15.29{\mu}m$. The chromosome complements comprised of eight pairs of metacentrics and one pair of submetacentric. Bleached DAPI Band analysis revealed a heterochromatin region, covering 28.6% to 50.4% each chromosomes. 5S and 45S rDNA sequences were located on two and three pairs of chromosomes, respectively. The centromere retrotransposon of Brassica (CRB) is a major component in Brassica related species that has been maintained as a common centromere component. CRB signals were detected on the centromere and pericentromeric region of R. sativus 'Wonkyo 10039' and three basic Brassica species (B. rapa, B. nigra, and B. oleracea). These results will provide a valuable background for physical mapping and elucidation of the evolutionary relationship among the Brassica related species.

Comparative Analysis of the Community of Culturable Bacteria Associated with Sponges, Spirastrella abata and Spirastrella panis by 16S rDNA-RFLP (16S rDNA-RFLP에 의한 Spirastrella abata와 Spirastrella panis 해면에 서식하는 배양가능한 공생세균 군집의 비교)

  • Cho, Hyun-Hee;Park, Jin-Sook
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.155-162
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    • 2009
  • A cultivation-based approach was employed to compare the culturable bacterial diversity associated with two phylogenetically closely related marine sponges, Spirastrella abata and Spirastrella panis, which have geologically overlapping distribution patterns. The bacteria associated with sponge were cultivated using MA medium supplemented with 3% sponge extracts. Community structures of the culturable bacteria of the two sponge species were analyzed with PCR-RFLP (restriction fragment length polymorphism) based on 16S rDNA sequences. The RFLP fingerprinting of 16S rDNA digested with HaeIII and MspI, revealed 24 independent RFLP types, in which 1-5 representative strains from each type were partially sequenced. The sequence analysis showed >98.4% similarity to known bacterial species in public databases. Overall, the microbial populations of two sponges investigated were found to be the members of the classes; Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria. The Alphaproteobacteria were predominant in the bacterial communities of the two sponges. Gammaproteobacteria represented 38.5% of bacterial community in S. abata. Whereas only 1.6% of this class was present in S. panis. Bacillus species were dominat in S. panis. Bacillus species were found to be 44.3% of bacterial species in S. panis, while they were only 9.7% in S. abata. It is interesting to note that Planococcus maritimus (8.1%, phylum Firmicutes) and Psychrobacter nivimaris (28.9%, phylum Gammaproteobacteria) were found only in S. abata. This result revealed that profiles of bacterial communities from the sponges with a close phylogenetic relationship were highly species-specific.

The complete plastid genome and nuclear ribosomal transcription unit sequences of Spiraea prunifolia f. simpliciflora (Rosaceae)

  • Jeongjin CHOI;Wonhee KIM;Jee Young PARK;Jong-Soo KANG;Tae-Jin YANG
    • Korean Journal of Plant Taxonomy
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    • v.53 no.1
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    • pp.32-37
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    • 2023
  • Spiraea prunifolia f. simpliciflora Nakai is a perennial shrub widely used for horticultural and medicinal purposes. We simultaneously obtained the complete plastid genome (plastome) and nuclear ribosomal gene transcription units, 45S nuclear ribosomal DNA (nrDNA) and 5S nrDNA of S. prunifolia f. simpliciflora, using Illumina short-read data. The plastome is 155,984 bp in length with a canonical quadripartite structure consisting of 84,417 bp of a large single-copy region, 18,887 bp of a short single-copy region, and 26,340 bp of two inverted repeat regions. Overall, a total of 113 genes (79 protein-coding genes, 30 tRNAs, and four rRNAs) were annotated in the plastome. The 45S nrDNA transcription unit is 5,848 bp in length: 1,809 bp, 161 bp, and 3,397 bp for 18S, 5.8S, and 26S, respectively, and 261 bp and 220 bp for internal transcribed spacer (ITS) 1 and ITS 2 regions, respectively. The 5S nrDNA unit is 512 bp, including 121 bp of 5S rRNA and 391 bp of intergenic spacer regions. Phylogenetic analyses showed that the genus Spiraea was monophyletic and sister to the clade of Sibiraea angustata, Petrophytum caespitosum and Kelseya uniflora. Within the genus Spiraea, the sections Calospira and Spiraea were monophyletic, but the sect. Glomerati was nested within the sect. Chamaedryon. In the sect. Glomerati, S. prunifolia f. simpliciflora formed a subclade with S. media, and the subclade was sister to S. thunbergii and S. mongolica. The close relationship between S. prunifolia f. simpliciflora and S. media was also supported by the nrDNA phylogeny, indicating that the plastome and nrDNA sequences assembled in this study belong to the genus Spiraea. The newly reported complete plastome and nrDNA transcription unit sequences of S. prunifolia f. simpliciflora provide useful information for further phylogenetic and evolutionary studies of the genus Spiraea, as well as the family Rosaceae.

Karyotype Analyses of a Rice Cultivar 'Nakdong' and its Four Genetically Modified Events by Conventional Staining and Fluorescence in situ Hybridization

  • Jeon, Eun Jin;Ryu, Kwang Bok;Kim, Hyun Hee
    • Korean Journal of Breeding Science
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    • v.43 no.4
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    • pp.252-259
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    • 2011
  • Conventional staining and fluorescence in situ hybridization (FISH) karyotypes of the non-genetically modified (GM) parental rice line, 'Nakdong' (Oryza sativa L. japonica), and its four GM rice lines, LS28 (event LS30-32-20-1), Cry1Ac1 (event C7-1-9-1), and LS28 ${\times}$ Cry1Ac1 (events L/C1-1-3-1 and L/C1-3-1-1) were analyzed using 5S and 45S rDNAs as probes. Both parental and transgenic lines were diploids (2n=24) with one satellite chromosome pair. The lengths of the prometaphase chromosomes ranged from 1.50 to $6.30{\mu}m$. Four submetacentric and eight metacentric pairs comprised the karyotype of 'Nakdong' and its four GM lines. One pair of 5S rDNA signals was detected near the centromeric region of chromosome g in both the parental and transgenic lines. The 45S rDNA signals were detected on the secondary constrictions of the satellite chromosome pair in both the parental and transgenic lines. There was no significant difference in chromosome size, length, and composition between 'Nakdong' and its four GM lines. This research was conducted as a preliminary study for chromosomal detection of transgenes in GM rice lines and would be useful for their breeding programs.

Cytogenetic Analyses of Angelica Plants Using Feulgen Staining and Multicolor Fluorescence in Situ Hybridization (Multicolor FISH와 Feulgen 염색법을 이용한 Angelica속 식물의 세포유전학적 분석)

  • Koo, Dal-Hoe;Kim, Soo-Young;Bang, Kyong-Hwan;Seong, Nak-Sul;Bang, Jae-Wook
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.123-127
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    • 2003
  • Karyotype analysis and chromosomal localization of 5S and 45S rDNAs using multi-color fluorescence in situ hybridization (McFISH) technique were carried out in two Angelica species. The numbers of diploid chromosomes were the same in two same in two species as 2n=22, however the lengths of chromosomes were varied from 4.25 to 6.50 ${\mu}{\textrm}{m}$ in A gigas and 4.95 to 8.50 ${\mu}{\textrm}{m}$ in A acutiloba. The chromosomes of A. gigas were composed of five metacentric and six submetacentric pairs, while those of A. acutiloba were six metacentic, one submetacentric and four subtelocentric paris. In FISH experiments, the numbers and size of 45S rDNA signals were varied between two species, however dach signal of the 5S rDNA was observed in two species.

Heterogeneity Analysis of the 16S rRNA Gene Sequences of the Genus Vibrio (Vibrio 속 16S rRNA 유전자 염기서열의 이질성 분석)

  • Ki, Jang-Seu
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.430-434
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    • 2009
  • Bacterial 16S rRNA gene sequences have been widely used for the studies on molecular phylogeny, evolutional history, and molecular detections. Bacterial genomes have multiple rRNA operons, of which gene sequences sometimes are variable. In the present study, heterogeneity of the Vibrio 16S rRNA gene sequences were investigated. Vibrio 16S rRNA sequences were obtained from GenBank databases, considering the completion of gene annotation of Vibrio genome sequences. These included V. cholerae, V. harveyi, V. parahaemolyticus, V. splendidus, and V. vulnificus. Chromosome 1 of the studied Vibrio had 7~10 copies of the 16S rRNA gene, and their intragenomic variations were less than 0.9% dissimilarity (more than 99.1% DNA similarity). Chromosome 2 had none or single 16S rRNA gene. Intragenomic 16S rRNA genotypes were detected at least 5 types (V. vulnificus #CMCP6) to 8 types (V. parahaemolyticus #RIMD 2210633, V. harveyi #ATCC BAA-1116). These suggest that Vibrio has high heterogeneity of the 16S rRNA gene sequences.

Phylogeographic Messages Encoded in the rDNA of the Commercial Mushroom Zhenghonggu@ From Fujian, China

  • Chen, Yu H.;Chen, Peng D.;Chen, Liu Y.;Ma, Li Z.
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.45-45
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    • 2014
  • Individualities of precious health mushroom called Zhenghonggu@ from respective protections scattered among all main mountains of Fujian China were collected and recognized locally, then compared with Russula griseocarnosa. Their internal transcribed spacer (ITS) region (ITS1, ITS2 and 5.8S rDNA) of the nuclear rDNA were amplified, AMOVA analyzed, nested clade analyzed and then compared with the ITS sequences of relative Russula species from other regions of China to confirm the taxonomic status of Zhenghonggu$^@$ and its population structure. Total 23 haplotypes from different protections of Fujian can be clustered into three clades similar to the three lineages of Dahongjun$^@$ from southeastern China reported by Li et al. The geographic distribution characteristic of these three phylogeny clades may be closely coupled with the vegetation regionalization and/or the differences of coenosium construction of Fagaceae that is the host of Russula griseocarnosa. The correlation of taxonomy, phylogeny and geographical distribution of Russula are discussed.

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A Report of Eighteen Unrecorded Fungal Species in Korea (국내에서 분리된 미기록 진균 18종 보고)

  • Ahn, Geum Ran;Choi, Min Ah;Kim, Ji Eun;Seo, Eun Ji;Kim, Jun Young;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.45 no.4
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    • pp.292-303
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    • 2017
  • During a survey of culturable fungi in Korea, 18 unrecorded fungal species were isolated and identified from the indoor air of mushroom cultivation houses, the materials used for preparation of mushroom cultivation media, wild plants, and funitures. This study reports the descriptions of the 18 unrecorded fungal species: Aspergillus creber, Ceratocystis paradoxa, Colletotrichum spaethianum, Coniochaeta velutina, Coprinellus xanthothrix, Epicoccum sorghinum, Leptosphaeria rubefaciens, Myrothecium gramineum, Paraconiothyrium fuckelii, Penicillium erubescens, Penicillium melinii, Penicillium pulvillorum, Penicillium sabulosum, Penicillium turbatum, Pestalotiopsis portugalica, Pilidiella castaneicola, Rachicladosporium pini, and Umbelopsis nana. For all the identified species, the morphological characteristics including the features of colony formed on media, images of light microscopy, and molecular phylogenetic relationships based on nucleotide sequences of the internal transcribed spacer ribosomal DNA (ITS rDNA), 18S rDNA, 28S rDNA, ${\beta}-tubulin$ gene, calmodulin gene, and translation elongation factor gene were described.

Cytogenetical Study of Cynanchum wilfordii and Cynanchum auriculatum using Fluorescence In Situ Hybridization (FISH) (rDNA FISH를 이용한 큰조롱과 넓은잎큰조롱의 세포유전학적 연구)

  • Kim, Su Hyun;Kim, Woo Cheol;Kim, Hyun Hee;Heo, Kweon
    • Korean Journal of Medicinal Crop Science
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    • v.28 no.5
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    • pp.325-330
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    • 2020
  • Background: Dried tuberous roots of Cynanchum wilfordii are known to relieve menopause symptoms. However, the dried roots of C. wilfordii are morphologically similar to those of C. auriculatum, which makes it difficult to distinguish when used as a medicine. Various comparative studies have focused on chemical or molecular analysis of these roots. However, the differences between the two species at the cytogenetic level based on chromosome structure and composition remain to be elucidated. Methods and Results: For chromosome slides, the roots were fixed in 8-hydroxyquinoline, digested with enzyme mixture, and spread on slides. 5S and 45S rDNA were used as cytogenetic markers for the analysis of nuclear genomes by FISH. The chromosome number of the two species was 2n = 22, with a relatively short length, 1.13 ㎛ - 4.24 ㎛ and 1.00 ㎛ - 3.42 ㎛ with respect to each other. Both species represent one pair of 5S and 45S rDNA signal on chromosome 1, at the proximal region and peri-centromeric region, respectively. Conclusions: These preliminary cytogenetic data using FISH in C. wilfordii and C. auriculatum could be valuable for the comprehension of Cynanchum genome history.