• Title/Summary/Keyword: Molecular sequence analyses

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A Novel Rapid Fungal Promoter Analysis System Using the Phosphopantetheinyl Transferase Gene, npgA, in Aspergillus nidulans

  • Song, Ha-Yeon;Choi, Dahye;Han, Dong-Min;Kim, Dae-Hyuk;Kim, Jung-Mi
    • Mycobiology
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    • v.46 no.4
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    • pp.429-439
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    • 2018
  • To develop a convenient promoter analysis system for fungi, a null-pigment mutant (NPG) of Aspergillus nidulans was used with the 4'-phosphopantetheinyl transferase (PPTase) gene, npgA, which restores the normal pigmentation in A. nidulans, as a new reporter gene. The functional organization of serially deleted promoter regions of the A. nidulans trpC gene and the Cryphonectria parasitica crp gene in filamentous fungi was representatively investigated to establish a novel fungal promoter assay system that depends on color complementation of the NPG mutant with the PPTase npgA gene. Several promoter regions of the trpC and crp genes were fused to the npgA gene containing the 1,034-bp open reading frame and the 966-bp 3' downstream region from the TAA, and the constructed fusions were introduced into the NPG mutant in A. nidulans to evaluate color recovery due to the transcriptional activity of the sequence elements. Serial deletion of the trpC and crp promoter regions in this PPTase reporter assay system reaffirmed results in previous reports by using the fungal transformation step without a laborious verification process. This approach suggests a more rapid and convenient system than conventional analyses for fungal gene expression studies.

The phylogeographic history of amphitropical Callophyllis variegata (Florideophyceae, Rhodophyta) in the Pacific Ocean

  • Bringloe, Trevor T.;Macaya, Erasmo C.;Saunders, Gary W.
    • ALGAE
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    • v.34 no.2
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    • pp.91-97
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    • 2019
  • Chilean species of marine macroalgae with amphitropical distributions oftentimes result from introductions out of the Northern Hemisphere. This possibility was investigated using haplotype data in an amphitropical red macroalgae present in Chile, Callophyllis variegata. Published sequence records from Canada and the United States were supplemented with new collections from Chile (April 2014-November 2015). Specimens of C. variegata were amplified for the 5′ end of the cytochrome c oxidase subunit I gene (COI-5P) and the full length nuclear internal transcribed spacer region. Haplotype networks and biogeographic distributions were used to infer whether C. variegata was introduced between hemispheres, and several population parameters were estimated using IMa2 analyses. C. variegata displayed a natural amphitropical distribution, with an isolation time of approximately 938 ka between hemispheres. It is hypothesized that contemporary populations of C. variegata were established from a refugial population during the late Pleistocene, and may have crossed the tropics via rafting on buoyant species of kelp or along deep-water refugia coincident with global cooling, representing a rare case of a non-human mediated amphitropical distribution.

Alsidium oliveiranum sp. nov. (Rhodomelaceae, Rhodophyta), an overlooked species from the southwestern Atlantic based on morphology and DNA sequence data

  • Guimaraes, Silvia M.P.B.;Soares, Luanda Pereira;Fujii, Mutue Toyota;Diaz-Tapia, Pilar
    • ALGAE
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    • v.34 no.3
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    • pp.187-198
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    • 2019
  • The exploration of seaweed diversity in poorly studied habitats has often led to the discovery of new species. Sand-covered rocks are an example, as they received less attention than sand-free rocky intertidal habitats during seaweed diversity surveys in Brazil. In sand-covered rocks from Espirito Santo and Rio de Janeiro we found an alga whose morphology was unique among rhodomelacean species previously reported in Brazil. With the aim to clarify the taxonomic identity of this species we studied its morphology, as well as its phylogenetic relationships. Molecular analyses resolved this species in the genus Alsidium (tribe Alsidieae) and differed from sequenced congeners with divergences ${\geq}2.5$ and 4.2% in the rbcL and cox1 genes, respectively. Morphological characters were in agreement with the genus Alsidium, and differed from other species currently recognized in the genus. The species consisted of a basal crust and scarcely branched erect axes with seven pericentral cells covered by a continuous layer of cortical cells. Reproductive structures were formed on clusters of short determinate branches. Therefore, the new species A. oliveiranum is proposed based on morphological and molecular evidence. Our findings contribute to better understand the diversity of the tribe Alsidieae, which is particularly diverse in the Americas.

Identification and molecular characterization of doublesex and mab-3-related transcription factor(dmrt) in brackish water flea, Diaphanosoma celebensis, exposed to bisphenol analogs

  • Cho, Hayoung;Jeon, Min Jeong;Lee, Young-Mi
    • Korean Journal of Environmental Biology
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    • v.39 no.2
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    • pp.160-168
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    • 2021
  • Doublesex and mab-3 related transcription factor(dmrt) play crucial roles in sex determination and sex differentiation in vertebrates and invertebrates. Although dmrt genes have been identified in vertebrates, little is known about aquatic invertebrates. In this study, two dmrt genes, namely, Dc_dmrt93B and Dc_dmrt99B, were identified from brackish water flea, Diaphanosoma celebensis. Transcriptional changes were observed in the dmrt genes when the flea was exposed to bisphenol(BP), an endocrine disruptor. Sequence and phylogenetic analyses showed that both dmrt genes contained two conserved domains, namely, DM and DMA, closely clustered with those of Daphnia spp. Additionally, a significant increase in the Dc_dmrt99B mRNA expression level was observed upon exposure to intermediate concentrations of BP (bisphenol A>bisphenol S=bisphenol F, p<0.05), while the expression of Dc_dmrt93B mRNA was slightly modulated. These findings imply that the two dmrt genes may be involved in sex differentiation of D. celebensis. Furthermore, it was found that the ability of BP to modulate dmrt genes could affect development and reproduction. This study provides a basis for understanding the function of the dmrt genes and the molecular mode of action of BP in small crustaceans.

A new record of high temperature tolerance species, Pyropia kitoi com. nov.(Bangiaceae, Rhodophyta), from Korea

  • Dong Jin Kim;Paola Romero-Orozco;Gwan Woung Kim;Seong Hyeon Baek;Tae Oh Cho;Boo Yeon Won
    • Korean Journal of Environmental Biology
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    • v.41 no.3
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    • pp.223-228
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    • 2023
  • Neoporphyra kitoi Ma. Abe, N. Kikuchi, Tamaki, Tom. Sato, Murase, Fujiyoshi & Mas. Kobayashi has been known as an endemic species in Japan. Its high temperature tolerance suggests that it could be advantageous for cultivation. In this study, we collected it from the Ulleungdo island, Korea and transferred it into Pyropia for a new combination, identified as Pyropia kitoi(Ma. Abe, N. Kikuchi, Tamaki, Tom. Sato, Murase, Fujiyoshi & Mas. Kobayashi) D.J. Kim, T.O. Cho & B.Y. Won comb. nov. based on morphological and molecular analyses. Pyropia kitoi is also reported as a new record species in the list of Korean macroalgal flora. Although we didn't observe the emergence of new blades from the rhizoidal cells, which is a key character for this species, our molecular analysis of rbcL revealed that our samples from Korea were congruent with "Neoporphyra kitoi" from Japan and were nested within the clade of Pyropia. The gene sequence divergence between the Korean and Japanese samples was 0-0.2%.

Molecular detection of Borrelia theileri in cattle in Korea

  • Hyeon-Ji Hyung;Yun-Sil Choi;Jinho Park;Kwang-Jun Lee;Jun-Gu Kang
    • Parasites, Hosts and Diseases
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    • v.62 no.1
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    • pp.151-156
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    • 2024
  • Bovine borreliosis, caused by Borrelia theileri which is transmitted via hard tick bites, is associated with mild clinical symptoms, such as fever, lethargy, hemoglobinuria, anorexia, and anemia. Borrelia theileri infects various animals, such as cattle, deer, horses, goats, sheep, and wild ruminants, in Africa, Australia, and South America. Notably, no case of B. theileri infection has been reported in Korean cattle to date. In this study, 101 blood samples were collected from a Korean indigenous cattle breed, among which 1.98% tested positive for B. theileri via nested PCR. The obtained sequences exhibited high homology with B. theileri strains identified in other regions. Phylogenetic analysis of 16S rRNA confirmed the B. theileri group affiliation; however, flagellin B sequences exhibited divergence, potentially due to regional evolutionary differences. This study provides the first molecular confirmation of B. theileri infection in Korean livestock. Further isolation and nucleotide sequence analyses are necessary to better understand the presence of B. theileri strains in cows in Korea.

Comparative chloroplast genomics and phylogenetic analysis of the Viburnum dilatatum complex (Adoxaceae) in Korea

  • PARK, Jongsun;XI, Hong;OH, Sang-Hun
    • Korean Journal of Plant Taxonomy
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    • v.50 no.1
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    • pp.8-16
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    • 2020
  • Complete chloroplast genome sequences provide detailed information about any structural changes of the genome, instances of phylogenetic reconstruction, and molecular markers for fine-scale analyses. Recent developments of next-generation sequencing (NGS) tools have led to the rapid accumulation of genomic data, especially data pertaining to chloroplasts. Short reads deposited in public databases such as the Sequence Read Archive of the NCBI are open resources, and the corresponding chloroplast genomes are yet to be completed. The V. dilatatum complex in Korea consists of four morphologically similar species: V. dilatatum, V. erosum, V. japonicum, and V. wrightii. Previous molecular phylogenetic analyses based on several DNA regions did not resolve the relationship at the species level. In order to examine the level of variation of the chloroplast genome in the V. dilatatum complex, raw reads of V. dilatatum deposited in the NCBI database were used to reconstruct the whole chloroplast genome, with these results compared to the genomes of V. erosum, V. japonicum, and three other species in Viburnum. These comparative genomics results found no significant structural changes in Viburnum. The degree of interspecific variation among the species in the V. dilatatum complex is very low, suggesting that the species of the complex may have been differentiated recently. The species of the V. dilatatum complex share large unique deletions, providing evidence of close relationships among the species. A phylogenetic analysis of the entire genome of the Viburnum showed that V. dilatatum is a sister to one of two accessions of V. erosum, making V. erosum paraphyletic. Given that the overall degree of variation among the species in the V. dilatatum complex is low, the chloroplast genome may not provide a phylogenetic signal pertaining to relationships among the species.

Identification of a Rice Gene (Bph 1) Conferring Resistance to Brown Planthopper (Nilaparvata lugens Stal) Using STS Markers

  • Kim, Suk-Man;Sohn, Jae-Keun
    • Molecules and Cells
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    • v.20 no.1
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    • pp.30-34
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    • 2005
  • This study was carried out to identify a high-resolution marker for a gene conferring resistance to brown planthopper (BPH) biotype 1, using japonica type resistant lines. Bulked segregant analyses were conducted using 520 RAPD primers to identify RAPD fragments linked to the BPH resistance gene. Eleven RAPDs were shown to be polymorphic amplicons between resistant and susceptible progeny. One of these primers, OPE 18, which amplified a 923 bp band tightly linked to resistance, was converted into a sequence-tagged-site (STS) marker. The STS marker, BpE18-3, was easily detectable as a dominant band with tight linkage (3.9cM) to Bph1. It promises to be useful as a marker for assisted selection of resistant progeny in backcross breeding programs to introgress the resistance gene into elite japonica cultivars.

Structural Characteristics of the Putative Protein Encoded by Arabidopsis AtMTN3 Gene

  • Cheong, Jong-Joo;Kwon, Hawk-Bin;Kim, Minkyun
    • Journal of Applied Biological Chemistry
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    • v.44 no.3
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    • pp.125-130
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    • 2001
  • A putative protein encoded by Arabidopsis AtMTN3 gene, a homologue of Medicago truncatula MTN3, consists of 285 amino acid residues, and has a predicted molecular mass of 31.5 kDa and a calculated pI of 9.1. Primary amino acid sequence analyses have revealed that the protein contains seven putative transmembrane regions with N-terminus oriented to the outside of the membrane. The AtMTN3 protein shows overall 16.4% of amino acid identity with the rat GALR3 protein, known to be a G-protein-coupled receptor. The gene is present as a single copy in the Arabidopsis genome, and expressed in aerial parts but not in roots of Arabidopsis. Therefore, AtMTN3 appears not to be specifically involved in Rhizobium-induced nodule development, as was predicted for the MTN3 gene. These proteins possibly mediate signal transmission through G-protein-coupled pathways during general interactions between plants and symbiotic or pathogenic microbes.

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Phylogenetic analysiccccccccc of the genus Stemphylium based on elongation factor -1 alpha and calmodulin gene squences

  • Kong, D.W.;Cho, H.S.;Yu, S.H.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.117.2-117
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    • 2003
  • The importance and diversity of the genus Stemphylium highlights the need for accurate identification of species. However, many Stemphylium isolates have been misidentified due to the use of spore size as the only identifying character. Molecular phylogenetic analyses were performed on fifty-four isolates covering 9 Stemphylium species collected in Korea. Phylogenetic analysis of the translation elongation factor -1 alpha (EF-1) and the calmodulin gene sequence data showed that Stemphylium species were segregated into seven distinct groups, most of w hichcorrelated with species identified by morphology. Analysis of EF-1 in particular was useful for establishing well- supported relationships among the species of Stemphylium.

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