• Title/Summary/Keyword: 45S and 5S rDNA

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A cytogenetic study of Astragalus koraiensis Y. N. Lee (정선황기의 세포유전학적 연구)

  • Han, Sang Eun;Kim, Hyun-Hee;Heo, Kweon
    • Korean Journal of Plant Taxonomy
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    • v.43 no.2
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    • pp.139-145
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    • 2013
  • This study was carried out to determine the karyotype and chromosomal localizations of 45S and 5S rDNAs using FISH in Astragalus koraiensis. The somatic metaphase chromosome number of this species was 2n = 16 with basic chromosome number of x = 8. The karyotype of A. koraiensis was consisted of six pairs of median region chromosomes(chromosome 1, 3, 4, 5, 6, 8) and two pairs of submedian chromosomes(chromosome 2, 7). Based on the FISH, one pair of 45S rDNA site was detected on the centromeric region of chromosome 5. Whereas, two pair of 5S sites were detected on the short arm of chromosome 4 and centromeric region of chromosome 7, respectively. These are quite different patterns from A. membranaceus, A. membranaceus var. alpinus, and A. mongholicus. Although A. koraiensis is considered as Korean endemic species, therefore, it should be conducted out comparative FISH study with A. sikokianus and A. bhotanensis which are very similar to A. koraiensis morphologically.

Analysis of Chromosome Composition of Gastrodia elata Blume by Fluorescent in situ Hybridization using rDNA and Telomeric Repeat Probes (rDNA와 말단소체 반복서열 탐침을 이용한 천마의 FISH 염색체 조성 분석)

  • Zhou, Hui Chao;Park, Eung Jun;Kim, Hyun Hee
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.2
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    • pp.113-118
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    • 2018
  • Background: Gastrodia elata Blume is a saprophytic perennial plant in the Orchidaceae family, because of its agricultural and medicinal effectiveness, researchers focus on its genome and chemical components. However, cytogenetic information based on the chromosome structure and composition to construct chromosomal backbone for genome sequencing research and for the development and breeding of plants is very limited. Methods and Results: We determined the metaphase chromosome composition of the G. elata genome by fluorescence in situ hybridization (FISH) using 5S and 45S rDNAs and telomeric repeat probes. The nuclear genome of G. elata was organized into 2 n = 36, with relatively small ($2.71-5.50{\mu}m$)chromosomes that showed gradual decrease in size. Conglutination phenomenon was observed among the metaphase chromosomes, and it was distinguished from that in other plant metaphase chromosome spreads. One pair of signal was detected for each 5S and 45S rDNA in the pericentromeric region and interstitial region on the short arm of chromosomes 10 and 4, respectively, and telomeric DNA signals were detected in the terminal region of most chromosomes. Conclusions: To our knowledge, this is the first FISH chromosome composition result in G. elata and could be useful in more comprehensive molecular cytogenetic and genomic analyses as well as breeding programs of the medicinal plant G. elata.

Characterization of a Ligninase Producing Strain, Serratia marcescens HY-5 isolated from Sympetrum dopressiusculum (고추좀잠자리 (Sympetrum depressiusculum)로부터 분리한 리그닌 분해균주, Serratia marcescens HY-5의 특성)

  • Kim, Ki-Duck;Park, Doo-Sang;Shin, Dong-Ha;Han, Bo-Na;Oh, Hyun-Woo;Youn, Young-Nam;Park, Ho-Yong
    • Korean journal of applied entomology
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    • v.45 no.3 s.144
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    • pp.301-307
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    • 2006
  • A ligniolytic bacterial strain was isolated from the digestive tract of a red dragonfly, Sympetrum dopressiusculum. It was identified as a Serratia marcescens HY-5 by 16S rDNA sequence analysis and physiological and biochemical analysis. The isolated strain showed proportional increase of ligninolytic activity to the cell growth in the culture media which include lignin compounds. It showed about 25-45% decomposition of lignin compound by 48 hr incubation especially, showed effective decomposition of monomer lignin compounds, vanillin and guaiacol, and a dimer, dealkaline lignin. PCR amplification of 16S rDNA followed by denaturing gradient gel electrophoresis analysis showed high density of S. marcescens HY-5 in the gut of the S. depressiusculum at both gut samples which collected at different site.

The Effect of Antibiotics on the DNA Synthesis and Base Composition in Fungal Cells (진균류의 DNA 생합성 및 염기조성에 미치는 항생물질의 효과)

  • Park, Kyou-Yeon;Lee, Chong-Sam
    • The Korean Journal of Mycology
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    • v.22 no.4
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    • pp.366-377
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    • 1994
  • The base composition of DNA of Aspergillus phoenicis, Rhizopus acidus and Candida albicans treated with cycloheximide and nalidixic acid during the culture was analyzed to compare with the control. The contents of base in the DNA were inhibited by cycloheximide, 20.4% of adenine, 43.1% of thymine, 40.9% of cytosine, 35.3% of guanine, 32.2% of purine, and 42.7% of pyrimidine for A. phoenicis. In R. acidus, 34.2% of adenine, 42.1% of thymine, 38.0% of cytosine, 18.1% of guanine, 24.1% of purine and 40.0% of pyrimidine were depressed by cycloheximide. In the antibiotic treatment of C. albicans, 58.3% of adenine, 58.5% of thymine, 58.1% of cytosine, 42.4% of guanine, 46.8% of purine and 58.8% of pyrimidine were inhibited to compare with the control. The nalidixic acid treatments were showed that, in A. phoenicis 41.6% of adenine, 47.1% of thymine, 59.3% of cytosine, 46.3% of guanine, 45.6% of purine and 57.2% of pyrimidine were inhibited. When R. acidus was treated with nalidixic acid, 59.1% of adenine, 54.7% of thymine, 35.3% of cytosine, 37.4% of guanine, 45.9% of purine and 44.9% of pyrimidine decreased. In treatment of nalidixic acid, the content of DNA was depressed 60.1% of adenine, 68.6% of thymine, 60.7% of cytosine, 40.0% of guanine, 45.8% of purine and 63.5% of pyrimidine for C. albicans In the DNA synthesis of three fungal cells, cycloheximide and nalidixic acid treatments were analyzed obviously that the biosynthesis of pyrimidine was depressed than that of purine. Therefore, it was showed that the DNA contents in the various fungal cells were inhibited remarkably in nalidixic acid treatment than cycloheximide.

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Karyotyping Analysis and Bicolor FISH of Pimpinella hallaisanensis, an Endemic to Jeju Island (제주특산 한라참나물(Pimpinella hallaisanensis)의 핵형분석과 Bicolor FISH)

  • Kim, Soo-Young;Kim, Chan-Soo;Tho, Jae-Hwa;Lee, Joongku
    • Korean Journal of Plant Taxonomy
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    • v.38 no.2
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    • pp.151-162
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    • 2008
  • Chromosome analysis using karyotyping and bicolor FISH were carried out in Pimpinella hallaisanensis which is one of the endemic plants in Jeju island of Korea. The somatic methaphase chromosomes number of this plant was 2n=2x=22 and the size of this chromosomes ranged from 3.58 to $5.82{\mu}m$. The chromosome complements consisted of two pairs of metacentrics (chromosomes 1 and 2), four pairs of submetacentrics (chromosomes 3, 4, 6 and 8) and five pairs of subtelocentrics (chromosomes 5, 7, 9, 10 and 11). Using bicolor FISH, three pairs of 5S and four pairs of 45S rDNA loci were observed. Two pairs of 5S rDNA signals were detected on the end of the long arm of chromosome 4 and one pair of them were observed between long arm end and centromere. Another 45S rDNA signals were detected on the end of short arm of chromosome 4, 6, 10 and 11, respectively. Hence, the chromosome number reexamined using both conventional staining and FISH methods was different from previous report.

Chromosome Analysis Using GISH and FISH of Interspecific Hybrids between Allium cepa L. and A. fistulosum L. (GISH와 FISH를 이용한 양파와 파간 종간교잡계통의 염색체 분석)

  • Kim, Cheol-Woo;Lee, Eul-Tai;Kim, Hwa-young;Choi, In-Hu;Bang, Jin-Ki;Koo, Dal-Hoe;Bang, Jae-Wook
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.468-473
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    • 2009
  • Fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) were used for chromosome analysis of hybrids (2n=16) between onion (Allium cepa L., 2n=2X=16) and welsh onion (A. fistulosum L., 2n=2X=16). 5S rDNA, 45S rDNA, and tandemly repeated DNA (TSD) sequence were used as probes for FISH analysis. A. fistulosum specific DNA probe of telomeric repeats and A. fistulosum DNA were used for GISH analysis. In the analysis of meiotic chromosome GISH revealed that hybrids have 7 bivalants and 2 univalents chromosome and 2 univalents were derived from A. fistulosum chromosomes. In somatic chromosomes of hybrid each 8 chromosomes were derived from A. cepa and A. fistulosum, respectively. FISH signal of 45S rDNA probe in A. fistulosum was detected at secondary constriction of chromosomes, while FISH signal in A. cepa was observed in both secondary constriction and telomere of chromosomes. TDS signals in A. fistulosum chromosomes were detected at all subtelomeric of 8 chromosomes and also in 2 pericentromeric of the chromosomes, whereas TDS signals in A. cepa were observed only in subtelomeric in all chromosomes. The pattern of TDS signal in hybrid chromosomes was similar to those of A. fistulosum chromosomes.

Identification of parasite DNA in common bile duct stones by PCR and DNA sequencing

  • Jang, Ji-Sun;Kim, Kyung-Ho;Yu, Jae-Ran;Lee, Soo-Ung
    • Parasites, Hosts and Diseases
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    • v.45 no.4
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    • pp.301-306
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    • 2007
  • We attempted to identify parasite DNA in the biliary stones of humans via PCR and DNA sequencing. Genomic DNA was isolated from each of 15 common bile duct (CBD) stones and 5 gallbladder (GB) stones. The patients who had the CBD stones suffered from cholangitis, and the patients with GB stones showed acute cholecystitis, respectively. The 28S and 18S rDNA genes were amplified successfully from 3 and/or 1 common bile duct stone samples, and then cloned and sequenced. The 28S and 18S rDNA sequences were highly conserved among isolates. Identity of the obtained 28S D1 rDNA with that of Clonorchis sinensis was higher than 97.6%, and identity of the 18S rDNA with that of other Ascarididae was 97.9%. Almost no intra-specific variations were detected in the 28S and 18S rDNA with the exception of a few nucleotide variations, i.e., substitution and deletion. These findings suggest that C. sinensis and Ascaris lumbricoides may be related with the biliary stoneformation and development.

Cytogenetic Mapping of Carthamus tinctorius L. with Tandemly Repeated DNA Sequences by Fluorescence in situ Hybridization

  • Mancia, Franklin Hinosa;Ju, Yoon Ha;Lim, Ki-Byung;Kim, Jung Sun;Nam, Sang Yong;Hwang, Yoon-Jung
    • Korean Journal of Plant Resources
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    • v.30 no.6
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    • pp.654-661
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    • 2017
  • Dual-color fluorescence in situ hybridization karyotype analysis was created using repetitive sequences including two types of rDNA repeats (45S and 5S rDNAs) and Arabidopsis-type telomere sequence repeats. The somatic metaphase cells of Carthamus tinctorius were observed as diploids (2n=2x=24). A symmetrical or slightly asymmetrical karyotype with seven pairs of metacentric and five pairs of submetacentric chromosomes was observed. The lengths of the somatic metaphase chromosomes ranged from 4.18 to $6.53{\mu}m$, with a total length of $60.71{\mu}m$. One locus of 45S rDNA was located on the pericentromeric regions of three pairs of chromosomes and the other pair was situated on the terminal regions of the short arms of a single pair of chromosomes. One locus of 5S rDNA was detected on the interstitial regions of the short arms of two pairs of chromosomes. Arabidopsis-type telomeric repeats were detected on the terminal regions of all pairs of chromosomes. Co-localization of loci between telomeric repeats and 45S rDNA was observed in a single pair of chromosomes. The results provide additional information for the existing physical mapping project of C. tinctorius and will also serve as a benchmark to a more intricate cytogenetic investigation of C. tinctorius and its related species.

The Diversity of Culturable Organotrophic Bacteria from Local Solar Salterns

  • Yeon, Sun-Hee;Jeong, Won-Jin;Park, Jin-Sook
    • Journal of Microbiology
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    • v.43 no.1
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    • pp.1-10
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    • 2005
  • We isolated and cultured bacteria inhabiting solar saltern ponds in Taean-Gun, Chungnam Province, Korea. All of the isolated 64 strains were found to be moderately halophilic bacteria, growing in a salt range of 2-20 %, with an optimal concentration of 5% salt. Bacterial diversity among the isolated halophiles was evaluated via RFLP analyses of PCR-amplified 16S rDNAs, followed by phylogenetic analysis of the partial 16S rDNA sequences. The combination of restriction enzyme digestions with HaeIII, CfoI, MspI and RsaI generated 54 distinct patterns. A neighbor-joining tree of the partial 16S rDNA sequences resulted in the division of the 64 strains into 2 major groups, 45 strains of ${\gamma}-Proteobacteria$ (70.3%) and 19 strains of Firmicutes (29.7%). The ${\alpha}-Proteobacteria$ and Cytophaga-Flavobacterium-Bacterioides groups, which were repeatedly found to exist in thalassohaline environments, were not represented in our isolates. The ${\gamma}-Proteobacteria$ group consisted of several subgroups of the Vibrionaceae (37.5%), Pseudoalteromonadaceae (10.9%), Halomonadaceae (7.8%), Alteromonadaceae (7.8%), and Idiomarinaceae (6.3%). Members of Salinivibrio costicola (29.7%) were the most predominant species among all of the isolates, followed by Halobacillus treperi (12.5%). Additionally, three new species candidates were found, based on similarities of the 16S rDNA sequences to those of previously published species.

FISH Karyotype and GISH Meiotic Pairing Analyses of a Stable Intergeneric Hybrid xBrassicoraphanus Line BB#5

  • Belandres, Hadassah Roa;Waminal, Nomar Espinosa;Hwang, Yoon-Jung;Park, Beom-Seok;Lee, Soo-Seong;Huh, Jin Hoe;Kim, Hyun Hee
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.83-92
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    • 2015
  • xBrassicoraphanus line BB#5, a new synthetic intergeneric hybrid between Brassica rapa L. ssp. pekinensis and Raphanus sativus L. var. rafiphera induced by N-methyl-N-nitroso-urethane mutagenesis in microspore culture, shows high seed fertility and morphological uniformity. Dual-color fluorescence in situ hybridization (FISH) using 5S and 45S rDNA probes and genomic in situ hybridization (GISH) using B. rapa genomic DNA probe were carried out to analyze the chromosome composition and the meiosis pairing pattern compared to its parental lines. The somatic chromosome complement is 2n = 38, which consists of 17 metacentric and two submetacentric chromosomes with lengths of 2.18 to $5.01{\mu}m$. FISH karyotype analysis showed five and eight pairs of 5S and 45S rDNA loci. GISH meiosis pairing analysis showed that 19 complete bivalents were most frequent and accounted for 42% of the 100 pollen mother cells examined. Based on chromosome number, size, morphology, rDNA distribution, and meiosis pairing pattern, both parental genomes of B. rapa and R. sativus appear to exist in xBrassicoraphanus line BB#5, demonstrating its genome integrity. Such stable chromosome constitutions and meiotic pairing patterns in somatic and meiotic cells are very rare in natural and synthetic intergeneric hybrids. Chromosomal studies and genetic and phenotypic changes in allopolyploids a re discussed. The results p resented h erein will b e usef ul f or f urther g enomic s tudy o f xBrassicoraphanus lines and their improvement as promising new breeding varieties.