• Title/Summary/Keyword: Ribosomal

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Molecular Variation and Distribution of Anopheles fluviatilis (Diptera: Culicidae) Complex in Iran

  • Naddaf, Saied Reza;Razavi, Mohammad Reza;Bahramali, Golnaz
    • Parasites, Hosts and Diseases
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    • v.48 no.3
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    • pp.231-236
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    • 2010
  • Anopheles fluviatilis James (Oiptera: Culicidae) is one of the known malaria vectors in south and southeastern Iran. Earlier ITS2 sequences analysis of specimens from Iran demonstrated only a single genotype that was identical to species Y in India, which is also the same as species T. We identified 2 haplotypes in the An. fluviatilis populations of Iran based on differences in nucleotide sequences of D3 domain of the 28S locus of ribosomal DNA (rDNA). Comparison of sequence data from 44 Iranian specimens with those publicly available in the Genbank database showed that all of the 288-D3 sequences from Kazeroun and Khesht regions in Fars Province were identical to the database entry representing species U in India. In other regions, all the individuals showed heterozygosity at the single nucleotide position, which identifies species U and T. It is argued that the 2 species may co-occur in some regions and hybridize; however, the heterozygosity in the 288-D3 locus was not reflected in ITS2 sequences and this locus for all individuals was identical to species T. This study shows that in a newly diverged species, like members of An. fluviatilis complex, a single molecular marker may not be sufficiently discriminatory to identify all the taxa over a vast geographical area. In addition, other molecular markers may provide more reliable information for species discrimination.

Comparison of Relationships in Infraspecies of Magnaporthe grisea Using DNA Sequence of Internal Transcribed Spacer II Region in Ribosomal DNA (도열병균(Magnaporthe grisea)의 Ribosomal DNA의 ITS II 부위 핵산 염기서열을 이용한 균주간 근연관계 비교)

  • 배신철;이신우;이인구;예완해;류진창
    • Korean Journal Plant Pathology
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    • v.12 no.1
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    • pp.91-98
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    • 1996
  • 벼도열병균 14개 균주와 벼 이외 화본과 식물 도열병균 12개 균주를 대상으로 rDNA의 ITS II 부위를 증폭하여 그들의 핵산 구조 차이를 분석함으로 도열병균 균주간 분류를 시도하였다. 5.8S rDNA의 3`-말단 부위와 28S rDNA의 5`-말단 부위의 sequence 중 5`-CCCGGGAATTCGCATCGATCGATCGAATGAAGA-ACGCAGC-3`와 5`-CCCGGGATCCTCCGCTTATT-GATATGC-3`를 이용하여 PCR 증폭을 하였을 때 벼도열병균 14개 균주는 동일한 길이의 단일 밴드를 형성하였으며 벼 이외 화본과 식물 도열병균에서는 레드톱 식물로부터 분리한 도열병균만이 나머지 균주보다 38bp가 더 큰 길이를 가진 밴드를 형성하였다. PCR로 증폭된 DNA를 HaeIII와 MspI 제한효소로 절단하였을 때 벼도열병균 레이스간에는 제한효소 절단에 의한 전기영동 밴드 형태 차이를 관찰할 수 없었으나, 벼 이외 화본과 식물 도열병균 12개 균주는 3군으로 구분할 수 있었다. 벼도열병균 90=054와 강아지풀에서 분리한 도열병균 G90-5, 기장에서 분리한 G88-4, 바랭이에서 분리한 G88-5 그리고 레드톱에서 분리한 RT 균주의 ITS II 부위의 DNA 염기서열 비교 분석에 의하면 G88-4와는 다른 HaeIII와 MspI 제한효소 위치를 가지고 있었기에 제한효소 절단에 의한 전기영동 형태가 상이하였다. 또한 RT균주는 HaeIII와 MspI위치가 존재하지 않았다.

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Three Intraspecific groups in Korean Isolates of Phytophthora drechsleri Based on PCR-RFLP of Ribosomal DNA (Ribosomal DNA의 PCR-RFLP에 의한 국내산 Phytophthora drechsleri의 3가지 종내그룹)

  • 홍승범;지형진;이승임;고승주;류진창;김인수
    • Korean Journal Plant Pathology
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    • v.14 no.5
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    • pp.519-525
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    • 1998
  • Intraspecific genetic diversity of Korean isolates of Phytophthora drechsleri was investigated based on PCR-RFLP of rDNA along with closely related species in the genus; P. cryptogea, P. melonis, P. erythroseptica, P. cinnamomi, P. cambivora and P. cactorum. Gene regions of nuclear small subunit and internal transcribed spacer (ITS) in rDNA were amplified with polymerase chain reaction and digested with 9 restriction enzymes. Phytophthora species was readily differentiated from each other based on the digestion patterns, however, P. cryptogea was not separable from some isolates of P. drechsleri. Twenty one isolates of P. drechsleri originated from 15 host plants were divided into three distinct groups designated as PdG1, PdG2 and PdG3, respectively. Four isolates in PdG1 were originated from green vegetables and tomato and nine isolates in PdG2 were mainly isolated from medicinal plants. The two groups showed 95.3% homology and four isolates of P. cyptogea came under the groups. However, Eight isolates in PdG3 collected from cucurbits were clearly differentiated from those of PdG1 and PdG2 by 66.5% homology, but completely matched with a Taiwan isolate of P. melonis. Results indicated that three distinct groups exist in Korean isolates of P. drechleri and each group has host preference. In addition, reclassification of the cucurbits isolates are reserved because of their distinct genetic characters from other intraspecific groups in P. drechsleri.

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A Case of Fatal Strongyloidiasis in a Patient with Chronic Lymphocytic Leukemia and Molecular Characterization of the Isolate

  • Kia, Eshrat Beigom;Rahimi, Hamid Reza;Mirhendi, Hossein;Nilforoushan, Mohammad Reza;Talebi, Ardeshir;Zahabiun, Farzaneh;Kazemzadeh, Hamid;Meamar, Ahmad Reza
    • Parasites, Hosts and Diseases
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    • v.46 no.4
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    • pp.261-263
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    • 2008
  • Strongyloides stercoralis is a human intestinal parasite which may lead to complicated strongyloidiasis in immunocompromised. Here, a case of complicated strongyloidiasis in a patient with chronic lymphocytic leukemia is reported. Presence of numerous S. stercoralis larvae in feces and sputum confirmed the diagnosis of hyperinfection syndrome in this patient. Following recovery of filariform larvae from agar plate culture of the stool, the isolate was characterized for the ITS1 region of ribosomal DNA gene by nested-PCR and sequencing. Albendazole therapy did not have cure effects; and just at the beginning of taking ivermectin, the patient died. The most important clue to prevent such fatal consequences is early diagnosis and proper treatment.

PCR-RFLP Analysis of Ribosomal DNA Intergenic Spacer Region in Fusarium section Liseola. (Fusarium section Liseola 균주들에서 rDNA Intergenic Spacer 부위의 PCR-RFLP 분석)

  • 이경은;최영길;민병례
    • Korean Journal of Microbiology
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    • v.38 no.1
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    • pp.7-12
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    • 2002
  • The intergenic spacer (IGS) region of the ribosomal DNA of species in Fusarium section Liseola was analyzed by amplification and subsequent digestion with several restriction enzymes. The length of the amplified IGS region was about 2.6 Kb in all strains except F.moniliforme 12 Which was about 2.9 Kb. The enzymes, EcoRI, HincII, SalI, HindIII, PstI and SmaI, digested the IGS region and nine haplotypes were identified among 11 strains. In the dendrogram based on PCR-RFLP of IGS region combined the results of section Liseola in this study and section Elegans in previous study, variation in the IGS appears to offer considerable potential to resolve intraspecific relationship as well as interspecies or intersection.

Morphological and Molecular Classifications of Genus Pholis

  • Lee, Sung-Hoon;Jang, Yo-Soon;Baik, Chung-Boo;Han, Kyeong-Ho;Myung, Jung-Goo;Lee, Jin-Hee;Choi, Sang-Duk;Kim, Seon-Jae;Kim, Jong-Oh;Hwang, Jae-Ho
    • Animal cells and systems
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    • v.13 no.4
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    • pp.453-460
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    • 2009
  • Morphological and molecular classifications were attempted in an effort to establish species-specific classifications of three species of the genus Pholis in Korea; these species were subjected to morphological and molecular methodologies using body measurements, RFLP, RAPD, and phylogenetic trees using the nucleotide sequences of mitochondrial 16S and 12S ribosomal DNAs, cytochrome c oxidase I, and cytochrome b. The data demonstrated that the three species of genus Pholis are distinct from each other, both morphologically and genetically.

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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Cultivation-Dependent and -Independent Characterization of Microbial Community Producing Polyhydroxyalkanoates from Raw Glycerol

  • Ciesielski, Slawomir;Pokoj, Tomasz;Klimiuk, Ewa
    • Journal of Microbiology and Biotechnology
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    • v.20 no.5
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    • pp.853-861
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    • 2010
  • High substrate costs decrease the profitability of polyhydroxyalkanoates (PHAs) production, and thus low-cost carbon substrates coming from agricultural and industrial residuals are tested for the production of these biopolymers. Among them, crude glycerol, formed as a by-product during biodiesel production, seems to be the most promising source of carbon. The object of this study was to characterize the mixed population responsible for the conversion of crude glycerol into PHAs by cultivation-dependent and -independent methods. Enrichment of the microbial community was monitored by applying the Ribosomal Intergenic Spacer Analysis (RISA), and the identification of community members was based on 16S rRNA gene sequencing of cultivable species. Molecular analysis revealed that mixed populations consisted of microorganisms affiliated with four bacterial lineages: ${\alpha}$, ${\gamma}$-Proteobacteria, Actinobacteria, and Bacteroides. Among these, three Pseudomonas strains and Rhodobacter sp. possessed genes coding for polyhydroxyalkanoates synthase. Comparative analysis revealed that most of the microorganisms detected by direct molecular analysis were obtained by the traditional culturing method.

Utility of taxon-specific molecular markers for the species identification of herbarium specimens: an example from Desmarestia japonica (Phaeophyceae, Desmarestiales) in Korea

  • Lee, Sang-Rae;Lee, Eun-Young
    • Fisheries and Aquatic Sciences
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    • v.21 no.3
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    • pp.8.1-8.6
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    • 2018
  • Desmarestia japonica (Phaeophyceae, Desmarestiales) was recently established from the Japanese ligulate Desmarestia and is morphologically similar to D. ligulata. This species has been reported only from Japan. However, the taxonomic reports based on additional regional distributions are needed to clarify this taxonomic entity and its species boundaries. Because Desmarestia species have restricted distributions in Korea, we reexamined herbarium specimens of D. ligulata deposited at the National Institute of Biological Resources (South Korea). To improve the amplification efficiency of the polymerase chain reaction and avoid contamination by the DNA of other organisms, we developed taxon-specific molecular markers suitable for DNA barcoding of Desmarestia species. Nuclear ribosomal small subunit RNA (18S rDNA) and mitochondrial cytochrome c oxidase 1 (cox1) regions were selected as target DNA. As a result, both were successfully isolated from herbarium specimens of D. japonica acquired over 10 years. These molecular markers provide useful genetic information for herbarium specimens for which conventional molecular analysis is challenging.

Phylogenetic Contributions of Partial 26S rDNA Sequences to the Tribe Helleboreae (Ranunculaceae)

  • Ro, Kyung-Eui;Han, Ho-Yeon;Lee, Sang-Tae
    • Animal cells and systems
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    • v.3 no.1
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    • pp.9-15
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    • 1999
  • Monophyly and intergeneric relationships of the tribe Helleboreae, sensu Tamura, and related genera were studied using a 1,100-bp segment at the 5'end of the 26S ribosomal RNA gene. Forty-one OTUs, including eight species of the Helleboreae, were either directly sequenced or obtained from previous publications. Data were analyzed using distance and discrete character methods to infer phylogenetic relationships among the included taxa. The inferred phylogeny did not support monophyly of either Helleboreae or Cimicifugeae whose members were intermixed in our inferred phylogeny. This result is congruent with our previous study, which recommended against finely subdividing, suprageneric higher taxa within the R-chromosome group (subfamily Ranuncluloideae, sensu lato) until more molecular data were accumulated. Our data convincingly suggest the presence of the following three monophyletic groups: the Cimicifuga group (the clade of Actaea, Cimicifuga, Souliea, Eranthis, Anemonopsis, and Beesia), the Trollius group (the clade of Trollius, Megaleranthis, Adonis), and a clade including Anemonopsis and Beesia. Our data also suggest that Trollius and Megaleranthis might be congeners and Eranthis a paraphyletic group.

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