• Title/Summary/Keyword: ITS2-rDNA region

Search Result 149, Processing Time 0.03 seconds

Transcriptional Activator Elements for Curtovirus C1 Expression Reside in the 3' Coding Region of ORF C1

  • Hur, Jingyung;Buckley, Kenneth J.;Lee, Sukchan;Davis, Keith R.
    • Molecules and Cells
    • /
    • v.23 no.1
    • /
    • pp.80-87
    • /
    • 2007
  • Beet curly top virus (BCTV) and Beet severe curly top virus (BSCTV), members of curtoviruses, encode seven open reading frames (ORFs) within a ~3 kb genome. One of these viral ORFs, C1, is known to play an important role in the early stage of viral infection in plants during initiation of viral DNA replication. We used promoter:: reporter (${\beta}$-glucuronidase) gene fusions in transgenic Arabidopsis to identify the putative promoter region of BCTV ORF C1. Unlike other geminiviruses, the intergenic region of BCTV was not sufficient to promote C1 expression in transgenic plants. When sequences extending into the coding region of C1 were tested, strong expression of the reporter protein was observed in vascular tissues of transgenic plants. This expression was not dependent on the presence of the intergenic regions or proximal 5' portions of the C1 coding region. Transgenic plants expressing a reporter gene under control of the putative complete C1 promoter were inoculated with virus to determine if any viral transcript affected C1 expression. Virus inoculated plants did not show any altered pattern or change in of reporter gene expression level. These results suggest that (1) important transcriptional activator elements for C1 expression reside in the 3' portion of C1 coding area itself, (2) C1 protein does not auto-regulate its own expression and (3) C1 expression of two curtoviruses is controlled differently compared to other geminiviruses.

Molecular Identification of a Sea Anemone (Cnidaria: Anthozoa: Actiniaria) Obtained in Gijang, Busan (부산 기장에서 채집된 말미잘의 분자생물학적 방법을 이용한 동정)

  • Yoo, Sang Joon;Kim, Do-Hyung
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.50 no.4
    • /
    • pp.447-452
    • /
    • 2017
  • In this study, we tried to identify a sea anemone collected from the coast of Gijang, Busan. The anemone was morphologically similar to species belonging to the genus Anthopleura, but its morphological characteristics did not allow for confirmed identification to species level. Multiple genes from mitochondrial cytochrome oxidase III, 12S and 16S rRNA, and nuclear 18S and 28S rRNA, were amplified for multilocus sequence typing (MLST) analysis using genomic DNA extracted from the sampled anemone and a different primer set. Based on the MLST analysis, the anemone obtained in this study was identified as Anthopleura artemisia. Also, the sequence of internal transcribed spacer-2 was most closely related to A. artemisia, indicating that this single region might be useful for anemone identification. This study shows significance of molecular identification for sea anemones, and will be helpful in studies of sea anemone identification using genotyping-by-sequencing.

Phylogenetic relationships of Lyophyllum decastes on the based of ITS region sequences and RAPD (잿빛만가닥버섯(Lyophyllum decastes)의 ITS 영역염기서열 및 RAPD에 의한 계통학적 유연관계 분석)

  • Woo, Sung-Mi;Park, Yong-Hwan;Yoo, Young-Bok;Shin, Pyung-Gyun;Jang, Kab Yeul;Jin, Yong-Ju;Sung, Jae-Mo
    • Journal of Mushroom
    • /
    • v.7 no.3
    • /
    • pp.98-104
    • /
    • 2009
  • Phylogenetic relationships of Hypsizygus mamoreus and Lyophyllum decastes artificial cultivated using ITS sequences and RAPD polymorphism have been analyzed. Based on ITS region sequences of 14 strains, we could divide into 2 group as group1 to Hypsizygus mamoreus and the control isolated group2 to Lyophyllum decastes were identified as the same species. Restrict analysis of rDNA ITS region which was amplified by PCR, 14 collected strains could be classified into 4 clusters. There was approximately 58% genetic similarity between cluster I(SPA 100, 101, 102) and cluster II(SPA 200, 208 and SPA 201, 202), 41% between cluster III(SPA 104, 103, 203) and cluster IV(SPA 204, 206, 207, 205) by BLAST analysis. RAPD polymorphisms were used to analyze the species diversity of artificially cultivated Lyophyllum decastes SPA 202. As a result, similarity between SPA 202 and SPA 203 was 75%, at the same time, similarity between SPA 202 and Pleurotus eryngii SPA 103 and SPA 104 was 65%.

  • PDF

New Species of the Genus Metschnikowia Isolated from Flowers in Indonesia, Metschnikowia cibodasensis sp. nov.

  • Sjamsuridzal, Wellyzar;Oetari, Ariyanti;Nakashima, Chiharu;Kanti, Atit;Saraswati, Rasti;Widyastuti, Yantyati;Ando, Katsuhiko
    • Journal of Microbiology and Biotechnology
    • /
    • v.23 no.7
    • /
    • pp.905-912
    • /
    • 2013
  • A novel species, Metschnikowia cibodasensis, is proposed to accommodate eight strains (ID03-$0093^T$, ID03-0094, ID03-0095, ID03-0096, ID03-0097, ID03-0098, ID03-0099, and ID03-0109) isolated from flowers of Saurauia pendula, Berberis nepalensis, and Brunfelsia americana in Cibodas Botanical Garden, West Java, Indonesia. The type strain of M. cibodasensis is ID03-$0093^T$ (= NBRC $101693^T$ =UICC $Y-335^T$ = BTCC-$Y25^T$). The common features of M. cibodasensis are a spherical to ellipsoidopedunculate shaped ascus, which contains one or two needle-shaped ascospores, and lyse at maturity. Asci generally develop directly from vegetative cells but sometimes from chlamydospores. The neighbor-joining tree based on the D1/D2 domain of nuclear large subunit (nLSU) ribosomal DNA sequences strongly supports that M. cibodasensis (eight strains) and its closest teleomorphic species, M. reukaufii, are different species by a 100% bootstrap value. The type strain of M. cibodasensis, ID03-$0093^T$, differed from M. reukaufii NBRC $1679^T$ by six nt (five substitutions and one deletion) in their D1/D2 region of nLSU rDNA, and by 18 nt (five deletions, four insertions, and nine substitutions) in their internal transcribed spacer regions of rDNA, respectively. Four strains representative of M. cibodasensis (ID03-$0093^T$, ID03-0095, ID03-0096, and ID03-0099) showed a mol% G+C content of $44.05{\pm}0.25%$, whereas that of M. reukaufii NBRC $1679^T$ was 41.3%. The low value of DNA-DNA homology (5-16%) in four strains of M. cibodasensis and M. reukaufii NBRC $1679^T$ strongly supported that these strains represent a distinct species.

DNA Polymorphism of Insulin-like Growth Factor-binding Protein-3 Gene and Its Association with Cashmere Traits in Cashmere Goats

  • Liu, Haiying;Liu, Chao;Yang, Guiqin;Li, Hui;Dai, Jin;Cong, Yuyan;Li, Xuejian
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.25 no.11
    • /
    • pp.1515-1520
    • /
    • 2012
  • Insulin-like growth factor binding protein-3 (IGFBP-3) gene is important for regulation of growth and development in mammals. The present investigation was carried out to study DNA polymorphism by PCR-RFLP of IGFBP-3 gene and its effect on fibre traits of Chinese Inner Mongolian cashmere goats. The fibre traits data investigated were cashmere fibre diameter, combed cashmere weight, cashmere fibre length and guard hair length. Four hundred and forty-four animals were used to detect polymorphisms in the hircine IGFBP-3 gene. A 316-bp fragment of the IGFBP-3 gene in exon 2 was amplified and digested with HaeIII restriction enzyme. Three patterns of restriction fragments were observed in the populations. The frequency of AA, AB and BB genotypes was 0.58, 0.33 and 0.09 respectively. The allelic frequency of the A and B allele was 0.75 and 0.25 respectively. Nucleotide sequencing revealed a C>G transition in the exon 2 region of the IGFBP-3 gene resulting in R158G change which caused the polymorphism. Least squares analysis revealed a significant effect of genotypes on cashmere weight (p<0.0001), cashmere fibre length (p<0.001) and hair length (p<0.05) of the animals. The effect of genotypes on cashmere fibre diameter was not statistically significant (p>0.05). The animals of AB and BB genotypes showed higher cashmere weight, cashmere fibre length and hair length than the animals possessing AA genotype. These results suggested that polymorphisms in the hircine IGFBP-3 gene might be a potential molecular marker for cashmere weight in cashmere goats.

Identification of Grovesinia moricola Causing Zonate Leaf Spots on Lespedeza cyrtobotrya in Korea (참싸리 겹둥근무늬병균 Grovesinia moricola 동정)

  • Park, Ji-Hyun;Jung, Bok-Nam;Lee, Sang-Hyun;Shin, Hyeon-Dong
    • The Korean Journal of Mycology
    • /
    • v.48 no.1
    • /
    • pp.69-74
    • /
    • 2020
  • In September 2017, a heavy damage by premature defoliation with the zonate leaf spots was observed in several shrubs of Lespedeza cyrtobotrya growing at Mt. Obongsan in Chuncheon, Korea. Numerous cone-shaped, white sporophores of a fungus were observed on lesions of the abaxial leaf surface. A similar fungus was isolated in September 2019 from the leaves of L. cyrtobotrya growing at Mt. Taegisan in Hoengseong, Korea. The morphological characteristics of the sporophores were consistent with those of Grovesinia moricola. The species identification was confirmed by sequencing the internal transcribed spacer (ITS) region of the ribosomal DNA from the two isolates (KACC48417 and KACC48934). The fungal pathogenicity was determined by an artificial inoculation in conditions of relative humidity and temperature of 100% and 15±2℃, respectively. This is the first report of association of G. moricola with L. cyrtobotrya in Korea.

Molecular Characterization of Various Trichomonad Species Isolated from Humans and Related Mammals in Indonesia

  • Kamaruddin, Mudyawati;Tokoro, Masaharu;Rahman, Md. Moshiur;Arayama, Shunsuke;Hidayati, Anggi P.N.;Syafruddin, Din;Asih, Puji B.S.;Yoshikawa, Hisao;Kawahara, Ei
    • Parasites, Hosts and Diseases
    • /
    • v.52 no.5
    • /
    • pp.471-478
    • /
    • 2014
  • Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.

Site-directed Mutagenesis Analysis Elucidates the Role of 223/227 Arginine in 23S rRNA Methylation, Which Is in 'Target Adenine Binding Loop' Region of ErmSF (위치 지정 치환 변이를 이용한 ErmSF의 '타깃 Adenine Binding Loop'을 형성하는 부위에 존재하는 223/227 Arginine 잔기의 23S rRNA Methylation 활성에서의 역할 규명)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
    • /
    • v.48 no.2
    • /
    • pp.79-86
    • /
    • 2012
  • ErmSF is one of the Erm family proteins which catalyze S-adenosyl-$_L$-methionine dependent modification of a specific adenine residue (A2058, E. coli numbering) in bacterial 23S rRNA, thereby conferring resistance to clinically important macrolide, lincosamide and streptogramin B ($MLS_B$) antibiotics. $^{222}FXPXPXVXS^{230}$ (ErmSF numbering) sequence appears to be a consensus sequence among the Erm family. This sequence was supposed to be involved in direct interaction with the target adenine from the structural studies of Erm protein ErmC'. But in DNA methyltarnsferase M. Taq I, this interaction have been identified biochemically and from the complex structure with substrate. Arginine 223 and 227 in this sequence are not conserved among Erm proteins, but because of the basic nature of residues, it was expected to interact with RNA substrates. Two amino acid residues were replaced with Ala by site-directed mutagenesis. Two mutant proteins still maintained its activity in vivo and resistant to the antibiotic erythromycin. Compared to the wild-type ErmSF, R223A and R227A proteins retained about 50% and 88% of activity in vitro, respectively. Even though those arginine residues are not essential in the catalytic step, with their positive charge they may play an important role for RNA binding.

Erysiphe lonicerigena sp. nov., a Powdery Mildew Species Found on Lonicera harae

  • In-Young Choi;Lamiya Abasova;Joon-Ho Choi;Jung-Hee Park;Hyeon-Dong Shin
    • Mycobiology
    • /
    • v.51 no.2
    • /
    • pp.67-71
    • /
    • 2023
  • A powdery mildew (Erysiphaceae) has been continuously collected on the leaves of Lonicera harae in the southern part of the Korean Peninsula, where this shrub is indigenous. Microscopic examination of the asexual morphs revealed that the current collections are differentiated from the all known Erysiphe species on Lonicera spp. by its longer conidiophores and longer conidia. Although the morphology of the chasmothecia is reminiscent of Erysiphe ehrenbergii and E. lonicerae, the specimens on L. harae differ from them in having smaller ascospores. A phylogenetic tree generated from a combined dataset of the internal transcribed spacer region and 28S rDNA gene sequences demonstrates that sequences obtained from three powdery mildew collections on L. harae clustered together as an independent species clade with high bootstrap values distant from other Erysiphe species on Lonicera, representing a species of its own. Based on morphological differences and molecular-phylogenetic results, the powdery mildew on L. harae is proposed as a new species, Erysiphe lonicerigena, and the holomorph of the fungus is described and illustrated in this study.

Characterization of a Novel Alga-Lytic Bacterium, Acidovorax temperans AK-05, Isolated from an Eutrophic Lake for Degradation of Anabaena cylindrica (부영양 호수에서 분리한 Acidovorax temperans AK-05의 Anabaena cylindrica 분해 특성)

  • Kim, Jeong-Dong;Han, Myung-Soo
    • Korean Journal of Ecology and Environment
    • /
    • v.37 no.2 s.107
    • /
    • pp.241-247
    • /
    • 2004
  • Isolation and identification of alga-lytic bacteria were carried out. Fifteen isolates of alga-lytic bacteria were screened by the double layer method using A. cylindrica NIES-19 as a sole nutrient and four isolates among them were compared with their alga-lytic activity. The isolate AK-05 exhibiting the highest alga-lytic activity was identified as Acidovorax temperans base on its 16S rDNA sequence. The culture supernatant of the isolate AK-05 was reliable for the alga-lytic. Alga-lytic activity assays of culture supernatant revealed that the major substances for alga-lytic activity were non-proteins and heat stable. The highest alga-Iytic activity was practical under alkaline conditions and at 25${\sim}$$30^{\circ}C$. It is indicating an advantage for the application of water blooms by cyanobacteria in eutrophic lakes where the pH is generally in alkaline region.