• 제목/요약/키워드: ITS rDNA sequences

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Comparative Analysis of Acanthopanax senticosus Harms from Korea, China and Russia Based on the ITS Sequences of Nuclear Ribosomal DNA (ITS 염기서열분석에 의한 한국산, 중국산 및 러시아산 가시오갈피의 유연관계 분석)

  • Han Hyo-Shim;Kim Doo-Young;Lee Kab-Yeon;Park Wan-Geun;Cho In-Kyung;Jung Jae-Sung
    • Korean Journal of Plant Resources
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    • v.19 no.1
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    • pp.54-58
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    • 2006
  • The genetic analyses of Acanthopanax senticosus Harms from Korea, China and Russia, were made by comparing the internal transcribed spacer (ITS) sequences of the nuclear ribosomal DNA. The ITS region of A. senticosus was amplified by polymerase chain reaction (PCR) using the universal primers and then directly sequenced. The length of the ITS region including 162 bp 5.85 rRNA gene ranged from 608 bp (for Korean and Chinese) to 611 bp (for Russian). The G+C content of ITS region were 60.20% for Korean and Chinese plants and 60.06% for Russian plants. Sequence comparisons indicated that ITS regions of A. senticosus from Korea and China were identical, whereas the ITS sequence of A. senticosus from Russia showed 99.2% homology with the plants from Korea. Variation in sequences were attributable to 5 bp substitution such as transversion or insertion events. These results suggested that A. senticosus Harms from Korea and China were closely related in phylogenetic relationship compared to Russian. In addition, A. senticosus Harms were more similar to Kalopanax pictus than A. sessiliflorus in their ITS sequences.

A Gene Encoding $\beta$-amylase from Saprolegnia parasitica and Its Expression in Saccharomyces cerevisiae

  • Kim, Hee-Ok;Park, Jeong-Nam;Shin, Dong-Jun;Lee, HwangHee Blaise;Chun, Soon-Bai;Bai, Suk
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.529-533
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    • 2001
  • The ${\beta}$-Amylase cDNA fragment from the oomcete Saprolegnia parasitica was cloned by reverse transcription-polymerase chain reaction (RT-PCR) using degenerate oligonucleotide primers derived from conserved ${\beta}$-amylase sequences. The 5'and 3'regions of the $\beta$-amylase gene were amplified using the rapid amplification of cDNA ends (rACE) system. It consisted of an open reading frame of 1,350 bp for a protein of 450 amino acids. Comparison between the genomic and cDNA sequences revealed that the intron was not present in the coding region. The deduced amino acid sequence of the ${\beta}$-amylase gene had a 97% similarity to the ${\beta}$-amylase of Saprolegnia ferax, followed by 41% similarity to those of Arabidopsis thaliana, Hordeum vulgare, and Zea mays. The ${\beta}$-amylase gene was also expressed in Saccharomyces cerevisiae by placing it under the control of the alcohol dehydrogenase gene (ADC1) promoter.

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Characterization of the Complete Mitochondrial Genome of Diphyllobothrium nihonkaiense (Diphyllobothriidae: Cestoda), and Development of Molecular Markers for Differentiating Fish Tapeworms

  • Kim, Kyu-Heon;Jeon, Hyeong-Kyu;Kang, Seokha;Sultana, Tahera;Kim, Gil Jung;Eom, Keeseon S.;Park, Joong-Ki
    • Molecules and Cells
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    • v.23 no.3
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    • pp.379-390
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    • 2007
  • We sequenced and characterized the complete mitochondrial genome of the Japanese fish tapeworm D. nihonkaiense. The genome is a circular-DNA molecule of 13607 bp (one nucleotide shorter than that of D. latum mtDNA) containing 12 protein-coding genes (lacking atp8), 22 tRNA genes and two rRNA genes. Gene order and genome content are identical to those of the other cestodes reported thus far, including its congener D. latum. The only exception is Hymenolepis diminuta in which the positions of trnS2 and trnL1 are switched. We tested a PCR-based molecular assay designed to rapidly and accurately differentiate between D. nihonkaiense and D. latum using species-specific primers based on a comparison of their mtDNA sequences. We found the PCR-based system to be very reliable and specific, and suggest that PCR-based identification methods using mtDNA sequences could contribute to the study of the epidemiology and larval ecology of Diphyllobothrium species.

Genetic Variation of Rice Populations Estimated Using nrDNA ITS Region Sequence

  • Wang, Dong;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.27 no.3
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    • pp.249-255
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    • 2014
  • The rice belonging to Oryza sativa is not only has significant economic importance, for it is the major source of nutrition for about 3 billion all around the world. But also plays a vital role as a model organism, because it has a number of advantages to be a model plant, such as efficient transformation system and small genome size. Many methods and techniques have been conducted to attempt to distinguish different Oryza sativa species, such as amplified fragment length polymorphism (AFLP), random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR) and so on. However, studies using sequence analysis of internal transcribed spacer (ITS), a region of ribosomal RNA has not been reported until now. This study was undertaken with an aim to understand the phylogenetic relationships among sixteen isolates of Oryza sativa collected from abroad and fifteen isolates collected from Korea, using ribosomal RNA (rRNA) internal transcribed spacer (ITS) sequences to compare the phylogeny relationships among different Oryza sativa species. The size variation obtained among sequenced nuclear ribosomal DNA (nrDNA) ITS region ranged from 515bp to 1000bp. The highest interspecific genetic distance (GD) was found between Sfejare 45 (FR12) and Anapuruna (FR15). Taebong isolate showed the least dissimilarity of the ITS region sequence with other thirty isolates. This consequence will help us further understanding molecular diversification in intra-species population and their phylogenetic analysis.

Isolation and Identification of Zinc-Enriched Yeast Saccharomyces cerevisiae FF-10 from the Tropical Fruit Rambutan (열대과일 Rambutan으로부터 아연 고함유 효모 Saccharomyces cerevisiae FF-10 분리 및 특성)

  • Cha, Jae-Young;Heo, Jin-Sun;Kim, Jung-Wook;Lee, Seon-Woo;Cho, Young-Su
    • Journal of Life Science
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    • v.18 no.4
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    • pp.447-453
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    • 2008
  • Zinc is an essential trace element in Human Being. Highly zinc containing yeast strain isolated from the tropical fruit, rambutan and the zinc concentration in this yeast strain was 306 ppm (30.6 mg%)per dry matter basis. This strain was found to be a rounded type, normal size, and multi-polar budding. Phylogenetic analysis using the ITS1-5.85 rDNA sequences from isolated strain is most similar to yeast Saccharomyces cerevisiae at the level of nucleotide sequence identity at 99%. This strain was produced alcohol by about 12% using fully colonized koji-rice with Aspergillus oryzae. In conclusion, the isolated strain was found to be closely related to the S. cerevisiae based on its morphological and physiological properties, and alcohol fermentation. The phylogenetic analysis of strain FF-10 using ITS 5.8S rDNA sequence data also supported the closely related to the S. cerevisiae. Accordingly, the isolated yeast was named as S. cerevisiae FF-10. Further studies on the best culture conditions for zinc production from zinc-enriched S. cerevisiae FF-10 are under investigation.

Internal Transcribed Spacer Barcoding DNA Region Coupled with High Resolution Melting Analysis for Authentication of Panax Species (DNA 바코딩과 고해상 융해곡선분석에 기반한 인삼속 식물의 종 판별)

  • Bang, Kyong Hwan;Kim, Young Chang;Lim, Ji Young;Kim, Jang Uk;Lee, Jung Woo;Kim, Dong Hwi;Kim, Kee Hong;Jo, Ick Hyun
    • Korean Journal of Medicinal Crop Science
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    • v.23 no.6
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    • pp.439-445
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    • 2015
  • Background : Correct identification of Panax species is important to ensure food quality, safety, authenticity and health for consumers. This paper describes a high resolution melting (HRM) analysis based method using internal transcribed spacer (ITS) and 5.8S ribosomal DNA barcoding regions as target (Bar-HRM) to obtain barcoding information for the major Panax species and to identify the origin of ginseng plant. Methods and Results : A PCR-based approach, Bar-HRM was developed to discriminate among Panax species. In this study, the ITS1, ITS2, and 5.8S rDNA genes were targeted for testing, since these have been identified as suitable genes for use in the identification of Panax species. The HRM analysis generated cluster patterns that were specific and sensitive enough to detect small sequence differences among the tested Panax species. Conclusion : The results of this study show that the HRM curve analysis of the ITS regions and 5.8S rDNA sequences is a simple, quick, and reproducible method. It can simultaneously identify three Panax species and screen for variants. Thus, ITS1HRM and 5.8SHRM primer sets can be used to distinguish among Panax species.

Revisiting the Parvilucifera infectans / P. sinerae (Alveolata, Perkinsozoa) species complex, two parasitoids of dinoflagellates

  • Jeon, Boo Seong;Nam, Seung Won;Kim, Sunju;Park, Myung Gil
    • ALGAE
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    • v.33 no.1
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    • pp.1-19
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    • 2018
  • Members of the family Parviluciferaceae (Alveolata, Perkinsozoa) are the well-known dinoflagellate parasitoids along with Amoebophrya ceratii species complex and parasitic chytrid Dinomyces arenysensis and contain six species across three genera (i.e., Parvilucifera infectans, P. sinerae, P. rostrata, and P. corolla, Dinovorax pyriformis, and Snorkelia prorocentri) so far. Among Parvilucifera species, the two species, P. infectans and P. sinerae, are very similar or almost identical each other morphologically and genetically, thereby make it difficult to distinguish between the two. The only main difference between the two species known so far is the number of sporangium wall (i.e., 2 layers in P. infectans vs. 3 layers in P. sinerae). During sampling in Masan bay, Korea during the spring season of 2015, the dinoflagellate Akashiwo sanguinea cells infected by the parasite Parvilucifera were observed and this host-parasite system was established in culture. Using this culture, its morphological and ultrastructural features with special emphasis on the variation in the number of sporangium wall over developmental times, were investigated. In addition, the sequences of rDNA regions and ${\beta}-tubulin$ genes were determined. The result clearly demonstrated that the trophocyte at 36 h was covered with 4 layers, and then outer layer of the sporocyte gradually degraded over time, resulting in wall structure consisting of two layers, with even processes being detached from 7-day-old sporangium with smooth surface, indicating that the difference in the number of layers seems not to be an appropriate ultrastructural character for distinguishing P. infectans and P. sinerae. While pairwise comparison of the large subunit rDNA sequences showed 100% identity among P. infectans / P. sinerae species complex, genetic differences were found in the small subunit (SSU) rDNA sequences but the differences were relatively small (11-13 nucleotides) compared with those (190-272 nucleotides) found among the rest of Parvilucifera species (P. rostrata and P. corolla). Those small differences in SSU rDNA sequences of P. infectans / P. sinerae species complex may reflect the variations within inter- strains of the same species from different geographical areas. Taken together, all morphological, ultrastructural, and molecular data from the present study suggest that they are the same species.

Phylogenetic Analysis of Ruminant Theileria spp. from China Based on 28S Ribosomal RNA Gene

  • Gou, Huitian;Guan, Guiquan;Ma, Miling;Liu, Aihong;Liu, Zhijie;Xu, Zongke;Ren, Qiaoyun;Li, Youquan;Yang, Jifei;Chen, Ze;Yin, Hong;Luo, Jianxun
    • Parasites, Hosts and Diseases
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    • v.51 no.5
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    • pp.511-517
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    • 2013
  • Species identification using DNA sequences is the basis for DNA taxonomy. In this study, we sequenced the ribosomal large-subunit RNA gene sequences (3,037-3,061 bp) in length of 13 Chinese Theileria stocks that were infective to cattle and sheep. The complete 28S rRNA gene is relatively difficult to amplify and its conserved region is not important for phylogenetic study. Therefore, we selected the D2-D3 region from the complete 28S rRNA sequences for phylogenetic analysis. Our analyses of 28S rRNA gene sequences showed that the 28S rRNA was useful as a phylogenetic marker for analyzing the relationships among Theileria spp. in ruminants. In addition, the D2-D3 region was a short segment that could be used instead of the whole 28S rRNA sequence during the phylogenetic analysis of Theileria, and it may be an ideal DNA barcode.

Diversity of Endophytic Fungi Associated with Roots of Calanthe discolor and Cephalanthera longibracteata in Korea (새우난초와 은대난초 뿌리에서 분리한 내생균의 다양성)

  • Lee, Bong-Hyung;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.427-435
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    • 2018
  • In this study, endophytic fungi were isolated from the surface-sterilized roots of Calanthe discolor and Cephalanthera longibracteata collected from the Chungnam, Jeju, Kyungnam and Chungbuk provinces in Korea. The morphological characteristics of the obtained isolates were examined and their sequences of the internal transcribed spacer (ITS) rDNA region were analyzed using the ITS1F and ITS4 primers for species identification. Leptodontidium orchidcola showed the highest species abundance and frequency among the isolated endophytic fungi. Additionally, the community analysis revealed a high specificity between the host plants and the endophytic fungal species.

Occurrence of Gray Mold Caused by Botrytis cinerea on Common Fig in Korea (Botrytis cinerea에 의한 무화과 잿빛곰팡이병)

  • Cheong, Seong-Soo;Choi, In-Young;Lee, Wang-Hyu
    • The Korean Journal of Mycology
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    • v.41 no.1
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    • pp.38-41
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    • 2013
  • In 2010 and 2011, gray mold was found on common fig (Ficus carica) fruit grown at the research field of Jeollabuk-do Agricultural Research and Extension Services, Korea. Gray mold symptoms on common fig fruit mainly occurred after harvest season until December. The typical symptom included brown water-soaked rot and fruit decay. The diseased fruit was covered by gray to brown colored conidiophore and conidia. The conidiophores were tree shape and measured $15-33{\times}2{\mu}m$. Conidia on conidiophore were ellipsoidal or lemon shape, colorless, single cell, and measured $7.3-14.6{\times}6.8-11.1{\mu}m$. The nucleotide sequences of the rDNA ITS region obtained from the pure culture of the gray mold on common fig were 100% similar to the sequences of the GenBank accession number HQ171052, EU519210, HQ171053, FN812726, HM849615, and EU563120 of B. cinerea isolates. In phylogenetic tree, the representative isolate was placed within same clade of B. cinerea. Based on the morphological characteristics and analysis of rDNA ITS sequence data, the fungus was identified as B. cinerea.