• Title/Summary/Keyword: 5.8S rDNA

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Secondary Structure of the Ribosomal Internal Transcribed Spacer (ITS) Region of Hypsizygus marmoreus (느티만가닥버섯의 ITS (internal transcribed spacer) 영역의 2차구조 분석)

  • Woo, Ju-Ri;Yoon, Hyeokjun;You, Young-Hyun;Lee, Chang-Yun;Kong, Won-Sik;Kim, Jong-Guk
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1260-1266
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    • 2013
  • The ribosomal DNA (rDNA) clusters of Hypsizygus marmoreus 3-10 and H. marmoreus 1-1 were analyzed in this study. The small subunit (SSU) and intergenic spacer 2 (IGS 2) was partially sequenced. The internal transcribed spacer 1 (ITS 1), 5.8S, internal transcribed spacer 2 (ITS 2), large subunit (LSU), intergenic spacer 1 (IGS 1), and 5S were completely sequenced. The rDNA clusters of H. marmoreus 3-10 and H. marmoreus 1-1 were 7,049 bp in length. The sequence of SSU rDNA, which corresponded to 18S rDNA, was 1,796 bp in length, and the sequence of LSU rDNA, which corresponded to 28S rDNA, was 3,348 bp in length. The ITS region that variable region and IGS region that non-transcribed spacer was 462 bp and 1,290 bp in length. The sequence of 5.8S rDNA and 5S rDNA was 153 bp and 43 bp in length, respectively. The 17 bp of the rDNA cluster in the H. marmoreus 3-10 strain was different to that in the H. marmoreus 1-1 strain, with 2 bp in the SSU, 3 bp in the ITS, 9 bp in the LSU, and 3 bp in the IGS. The analysis of the secondary structure revealed that the ITS regions of H. marmoreus 3-10 and H. marmoreus 1-1 have five stem-loop structures. Interestingly, among these structures, one different nucleotide sequence resulted in a different secondary structure in stem-loop V.

Cytogenetic Analyses of Astragalus Species (황기류 식물 3종의 세포유전학적 분석)

  • Kim, Soo-Young;Choi, Hae-Woon;Kim, Chan-Soo;Sung, Jung-Sook;Lee, Joong-Ku;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.4
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    • pp.250-254
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    • 2006
  • To elucidate cytogenetic differences, karyotype analysis and FISH (fluorescence in situ hybridization) with 45S and 5S rDNAs were carried out in the three Astragalas species: Astragalas membranaceus Bunge, A. membranaceus var. alpinus Nakai and A. mongholicus Bunge. The somatic metaphase chromosome numbers of all three species were 2n=2x=16 and the size of chromosomes ranged $2.19{\sim} 5.73\;{\mu}m$. The chromosome complement of A. membranaceus consisted of each four pairs of metacentrics (chromosomes 3,4,6 and 7) and submetacentrics (chromosomes 1,2,4 and 8). In A. membranaceus var. alpinus, the chromosome complement consisted of two pairs of metacentrics (chromosomes 4 and 8) and six pairs of submetacentrics (chromosomes 1,2,3,5,6 and 7). A. mongholicus had three pairs of metacentrics (chromosomes 6,7 and 8) and five pairs of submetacentrics (chromosomes 1,2,3,4 and 5). Using bicolor-FISH, one pair of 45S and 5S rDNA signals could be detected on the centromeric regions of chromosomes 8 and 7 of A. membranaceus and A. mongholicus, respectively. In contrast, A, membranaceus var. alpinus had one pair of 45S signals on the centromeric region of chromosome 8 and two pairs of 5S rDNA signals on the short arms of chromosomes 7 and 8.

Intrageneric Relationships of Trichoderma Based on Internal Transcribed Spacers and 5.8S rDNA Nucleotide Sequences

  • Kim, Gi-Young;Lee, Goang-Jae;Ha, Myung-Gyu;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.28 no.1
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    • pp.11-16
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    • 2000
  • The nucleotide sequences of the internal transcribed spacer (ITS) regions of the ribosomal DNA including the 5.8S ribosomal RNA gene (rDNA) have been determined for 11 species in order to analyze their intrageneric relationships. The total length of these sequences ranged from 530 nucleotides for Trichoderma reesei KCTC 1286 to 553 nucleotide for Trichoderma koningii IAM 12534. Generally speaking, the length of ITS1 region was about 30 nucleotides longer than that of the ITS2 region. Also, the sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites were also found. Thus, a neighbor-joining tree was constructed using the full sequence data of the ITS regions and the 5.8S rDNA. The Trichoderma genus used to be grouped on the basis of the morphological features and especially the shape of phialides needs to be reexamined. The phylogenetic tree displayed the presence of monophylogeny in the species of Trichoderma. Therefore, it was difficult to distinguish the intrageneric relationships in the Trichoderma genus.

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Phylogenetic and Chemical Analyses of Cirsium pendulum and Cirsium setidens Inhabiting Korea (국내에 자생하는 큰엉겅퀴와 고려엉겅퀴의 분자유전학적 및 화학적 분석)

  • Yoo, Sun-Kyun;Bae, Young-Min
    • Journal of Life Science
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    • v.22 no.8
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    • pp.1120-1125
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    • 2012
  • Cirsium pendulum plants were collected from Hongcheon, Pyeongchang, Wonju, Yangyang in Kangwondo, Gapyeong in Gyeongkido, and Choongju in Choongcheongbukdo. Cirsium setidens plants were collected from Taebaek in Kangwondo and Bonghwa in Kyeongsangbukdo. Genomic DNA was prepared from those plants and used for the amplification of 18S rDNA, ITS1, 5.8S rDNA, ITS2, and part of 28S rDNA. The PCR products were sequenced, and the sequence was deposited in the GenBank. The comparison of those sequences has revealed that the rDNA sequences are identical for all six C. pendulum plants, but that the ITS1 and ITS2 sequences contain variable nucleotides. The two C. setidens plants had different nucleotides in 18S rDNA, ITS1, and ITS2. The comparison of the DNA sequences of C. pendulum and C. setidens collected in this study with C. pendulum of Hokkaido in Japan and C. japonicum of Anhui in China indicated that the plants of those three species are clearly divided into three distinct groups. The silymarin content of the collected plants was analyzed and turned out to be quite high. Therefore, it has been found that both C. pendulum and C. setidens plants are producing large amounts of silymarin, which has been reported to have various medicinal effects.

Cytogenetic Study of Maackia amurensis Rupr. & Maxim. and M. fauriei (Levl.) Takeda Using Karyotyping Analysis and the FISH Technique (핵형분석과 FISH 기술을 이용한 솔비나무와 다릅나무의 세포유전학적 연구)

  • Kim, Soo-Young;Kim, Chan-Soo
    • Korean Journal of Plant Taxonomy
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    • v.39 no.3
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    • pp.193-198
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    • 2009
  • Chromosome analysis using karyotyping and bicolor FISH were carried out for two Maackia species (M. fauriei and M. amurensis) found in Korea. The somatic metaphase chromosome number was 2n = 2x = 18 in both, and the size of these chromosomes ranged from 3.58 to $5.82{\mu}m$. The chromosome complements consisted of two pairs of metacentric (chromosomes 1 and 7), four pairs of submetacentrics (chromosomes 4, 6, 8 and 9) and three pairs of subtelocentrics (chromosomes 2, 3 and 5) in M. fauriei but, chromosomes 4 (subtelocentric) and 7 (submetacentric) of M. amurensis have different morphology. Using bicolor FISH, a pair of 45S rDNA loci were observed for both M. fauriei and M. amurensis, but the number and site of the 5S rDNA signal were different in the two species. M. fauriei has two pairs of 5S signals on chromosomes 7 and 8 but, M. amurensis has four paris on chromosomes 3, 4, 7 and 7. Hence, the 5S rDNA is a useful FISH for Maackia species.

Analysis of FISH patterns using 5S and 45S rDNAs in Codonopsis minima and C. lanceolata from Jeju Island (5S와 45S rDNA 유전자를 이용한 제주도산 애기더덕 (Codonopsis minima)과 더덕 (C. lanceolata)의 FISH 패턴 분석)

  • Kim, Soo-Young;Kim, Chan-Soo
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.3
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    • pp.186-190
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    • 2010
  • The chromosome number was identified and fluorescence in situ hybridization(FISH) mapping of 5S and 45S rDNAs were conducted for C. minima and C. lanceolata in the genus Codonopsis from Jeju island. In this study, we have confirmed that the somatic metaphase chromosome number determined as 2n=2x=16 was the same as the findings from the previous studies. While the conventional staining method makes it rather difficult to distinguish satellite chromosomes due to high degree of variability, FISH analysis produced the exact number and location of 5S and 45S rDNAs. Both species in the genus Codonopsis have a pair of 5S rDNA and their gene loci were observed on chromosome 3. Although two pairs of 45S rDNAs (one on chromosome 1 and the other on chromosome 8) were identified in both species, the 45S rDNA signals on chromosome 8 in C. minima were significantly weaker than those on chromosome 1. In addition, the 45S rDNA signals on chromosome 1 in C. lanceolata showed that the chromosome is non-homologus. In this study, we have determined cytogenetic characteristics of C. minima and C. lanceolata according to their gene replication patterns.

FISH Karyotype Analysis of Four Wild Cucurbitaceae Species Using 5S and 45S rDNA Probes and the Emergence of New Polyploids in Trichosanthes kirilowii Maxim

  • Waminal, Nomar Espinosa;Kim, Hyun Hee
    • Horticultural Science & Technology
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    • v.33 no.6
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    • pp.869-876
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    • 2015
  • Wild relative species of domesticated crops are useful genetic resources for improving agronomic traits. Cytogenetic investigations based on chromosome composition provide insight into basic genetic and genomic characteristics of a species that can be exploited in a breeding program. Here, we used FISH analysis to characterize the ploidy level, chromosome constitution, and genomic distribution o f 5S and 4 5S r ibosomal DNA (rDNA) in four wild Cucurbitaceae species, namely, Citrullus lanatus (Thunb.) Mansf. var. citroides L. H. Bailey (2n = 22), Melothria japonica Maxim. (2n = 22), Sicyos angulatus L. (2n = 24), and Trichosanthes kirilowii Maxim. (2n = 66, 88, 110 cytotypes), collected in different areas of Korea. All species were diploids, except for T. kirilowii, which included hexa-, octa-, and decaploid cytotypes (2n = 6x = 66, 8x = 88, and 10x = 110). All species have small metaphase chromosomes in the range of $2-5{\mu}m$. The 45S rDNA signals were localized distally compared to the 5S rDNA. C. lanatus var. citroides and M. japonica showed one and two loci of 45S and 5S rDNA, respectively, with co-localization of rDNA signals in one M. japonica chromosome. S. angulatus showed two co-localized signals of 5S and 45S rDNA loci. The hexaploid T. kirilowii cytotype showed five signals each for 45S and 5S rDNA, with three being co-localized. This is the first report of hexaploid and decaploid cytotypes in T. kirilowii. These results will be useful in future Cucurbitaceae breeding programs.

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.

A Phylogenetic Relationship between Foreign and Korean Strains of Flammulina velutipes Identified by rDNA-ITS Sequence Analysis (Flammulina velutipe의 국내 균주와 외래 균주 간의 ITS region을 이용한 계통학적 유연관계 분석)

  • Hwang, Gwang-Rip;Woo, Ju-Ri;Yoon, Hyeok-Jun;Lee, Chang-Yun;Lee, Sang-Han;Kong, Won-Sik;Kim, Jong-Guk
    • Journal of Life Science
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    • v.22 no.1
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    • pp.62-73
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    • 2012
  • This study was carried out to investigate the genetic relationship of Flammulina velutipes with other species. The ribosomal DNA cluster containing 4 rRNA genes from F. velutipes 4154 were sequenced. The length of the rDNA cluster sequence was estimated at 7,403 bp long and consisted of 1,806 bp of SSU rDNA, 245 bp of ITS 1 region, 159 bp of 5.8S rDNA, 308 bp of ITS 2 region, 3,402 bp of LSU rDNA, 1,400 bp of IGS 1 region, and 83 bp of 5S rDNA. The F. velutipes 4154 genes were contained in the rDNA cluster of F. velutipes in the order of SSU rDNA - ITS 1 - 5.8S rDNA - ITS 2 - LSU rDNA - IGS 1 - 5S rDNA. The phylogenetic relationships of 20 strains of Tricholomataceae and Physalacriaceae were analyzed by conducting distance analysis using the Neighbor-joining (NJ) method. The 20 strains used in this study were divided into three groups and the strains of the genus Flammulina were related very closely to strains of Physalacria bambusae.