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Internal Transcribed Spacer (internal + transcribed_spacer)

Distribution by Scientific Domains
Life Sciences87%
Medical Sciences5%
3 Other Domains8%
Distribution within Life Sciences
Evolution27%
Phycology18%
Plant Science13%
Plant Pathology12%
Microbial Ecology8%
Entomology6%
5 Other Subdomains16%

Kinds of Internal Transcribed Spacer

  • dna internal transcribed spacer
  • nuclear internal transcribed spacer
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  • nuclear ribosomal internal transcribed spacer
  • rdna internal transcribed spacer
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  • ribosomal dna internal transcribed spacer
  • ribosomal internal transcribed spacer
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    Terms modified by Internal Transcribed Spacer

  • internal transcribed spacer region
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  • internal transcribed spacer regions
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  • internal transcribed spacer sequence
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    Selected Abstracts

    Mycosphaerella species associated with leaf disease of Eucalyptus globulus in Ethiopia

    FOREST PATHOLOGY, Issue 4 2006
    Alemu Gezahgne
    Summary Eucalyptus spp. are among the most widely planted exotic trees in Ethiopia. Several damaging leaf pathogens are known from Eucalyptus spp. worldwide. Of these, Mycosphaerella spp. are among the most important, causing the disease known as Mycosphaerella leaf disease (MLD). Characteristic symptoms of MLD include leaf spot, premature defoliation, shoot and twig dieback. Recent disease surveys conducted in Ethiopian Eucalyptus plantations have revealed disease symptoms similar to those caused by Mycosphaerella spp. These symptoms were restricted to E. globulus trees growing in several localities in south, south western and western Ethiopia. The aim of this study was to identify the fungi associated with this disease. This was achieved by examining ascospore germination patterns, anamorph associations and sequence data from the Internal Transcribed Spacer (ITS) region of the rRNA operon, for representative isolates. Several different ascospore germination patterns were observed, suggesting that more than one species of Mycosphaerella is responsible for MLD on E. globulus in Ethiopia. Analysis of sequence data showed that three Mycosphaerella spp., M. marksii, M. nubilosa and M. parva were present. This is the first report of these three species from Ethiopia and represents a valuable basis on which to build further studies in the region. Résumé Les Eucalyptus comptent parmi les essences d'arbres exotiques les plus plantées en Ethiopie. Plusieurs pathogènes foliaires sont connus dans le monde pour occasionner des dégâts sur Eucalyptus. Parmi ceux-ci, les espèces de Mycosphaerella sont parmi les plus importantes, causant la maladie connue comme Maladie Foliaire àMycosphaerella (MFM, MLD en anglais). Les symptômes caractéristiques de la MFM comprennent des taches foliaires, une défoliation précoce et des dépérissements de pousses et de rameaux. Des campagnes de surveillance menées récemment dans les plantations éthiopiennes d'Eucalyptus ont révélé la présence de tels symptômes. Ces symptômes sont uniquement observés sur E. globulus dans plusieurs localités du sud, sud-ouest et ouest de l'Ethiopie. L'objectif de cette étude était d'identifier les champignons associés à cette maladie. Pour cela, des isolats représentatifs ont étéétudiés pour les modalités de germination des ascospores, les anamorphes associés ainsi que les données de séquence de la région ITS de l'opéron ADNr. Différentes modalités de germination des ascospores ont été observées, suggérant que plusieurs espèces de Mycosphaerella seraient associées à la MFM sur E. globulus en Ethiopie. L'analyse des données de séquence a montré la présence de 3 espèces : M. marksii, M. nubilosa et M. parva. Ceci constitue la première mention de ces 3 espèces en Ethiopie et une première étape pour envisager d'autres études dans cette région. Zusammenfassung Eucalyptus -Arten sind die am häufigsten angepflanzten exotischen Bäume in Äthiopien. An Eucalyptus kommen verschiedene Blattkrankheiten vor, wobei die Mycosphaerella -Arten als Verursacher der Mycosphaerella -Blattkrankheit (MLD) am bedeutendsten sind. Charakteristische Symtpome der MLD sind Blattnekrosen und vorzeitiger Blattfall sowie Trieb- und Zweigsterben. Bei der Inventur von Krankheiten in äthiopischen Eucalyptusplantagen wurden Symptome entdeckt, die denen von Mycosphaerella spp. ähnlich waren. Diese traten nur an E. globulus lokal in S-, SW- und W-Äthiopien auf. Ziel dieser Untersuchung war es, die damit assoziierten Pilze zu identifizieren. Hierzu wurde an repräsentativen Isolaten das Keimverhalten der Ascosporen, das Vorkommen von Anamorphen und die ITS-Sequenz des rRNA-Operons untersucht. Es wurden verschiedene Keimungstypen der Ascosporen beobachtet, was darauf schliessen liess, dass mehr als eine Mycosphaerella -Art für die Krankheit an E. globulus in Äthiopien verantwortlich ist. Anhand der Sequenzen wurden M. marksii, M. nubilosa und M. parva identifiziert. Dies ist der Erstnachweis für diese drei Arten in Äthiopien und eine Grundlage für weitere Studien. [source]

    A SYSTEMATIC STUDY OF GIGAR-TINACEAE FROM PACIFIC NORTH AMERICA BASED ON MOLECULAR AND MORPHOLOGICAL EVIDENCE

    JOURNAL OF PHYCOLOGY, Issue 2000
    J.R. Hughey
    Greater than 50 species of Gigartinaceae have been described from Pacific North America, about half of which are currently recognized. Although the family is treated extensively in the taxonomic literature, many of the species are still confused and a comprehensive revision is required. We sequenced the rbcL (RuBisCO) gene and ITS (Internal Transcribed Spacer) 1, 2, and 5.8S regions from a large number of recent collections and identified a discrete of number data sets. These were analysed in comparison with the morphological evidence for each of the taxa. Uncertain of the possibility that our operational taxonomic units may not correspond to the types, we developed a protocol for isolating PCR-friendly DNA from herbarium specimens, some reaching back as far as 1670. The DNA profiles of types and historically important specimens were compared to those for recently collected silica gel-dried and formalin-fixed material and assigned correct names. Species studied ranged from Alaska to Mexico and the Gulf of California and were compared to outgroup taxa from Pacific South America and the Southern Ocean. Particular attention was paid to variations in morphology as they relate to habitat with emphasis on the presence or absence of different morphological forms among sympatric and allopatric populations. We recognize 10 species in Chondracanthus (including one new combination and one new species) and 16 species in Mazzaella (including two new combinations and two new species). Finally, we tested a phylogenetic hypothesis inferred for the Gigartinaceae from rbcL sequences for congruence with one generated from ITS sequences. [source]

    Ericoid mycorrhizal fungi are common root inhabitants of non- Ericaceae plants in a south-eastern Australian sclerophyll forest

    FEMS MICROBIOLOGY ECOLOGY, Issue 2 2008
    Susan M. Chambers
    Abstract Fungi were isolated from the roots of 17 plant species from the families Apiaceae, Cunoniaceae, Cyperaceae, Droseraceae, Fabaceae-Mimosoideae, Lomandraceae, Myrtaceae, Pittosporaceae, Proteaceae and Stylidiaceae at a sclerophyll forest site in New South Wales, Australia. Internal transcribed spacer (ITS) restriction fragment length polymorphism (RFLP) and sequence comparisons indicated that the isolated fungi had affinities to a range of ascomycetes, basidiomycetes and zygomycetes. Four RFLP types had closest affinities to previously identified Helotiales ericoid mycorrhizal (ERM) or Oidiodendron spp. Isolates representing six RFLP types, which were variously isolated from all 17 plant species, formed ERM coils in hair root epidermal cells of Woollsia pungens (Ericaceae) under gnotobiotic conditions. Three of these isolates formed intercellular hyphae, intracellular hyphae and/or microsclerotia, which are typical of dark septate endophyte infection, in roots of Stylidium productum (Stylidiaceae), indicating an ability to form different types of association with roots of different hosts. Overall the data indicate that a broad range of plant taxa may act as repositories for ERM fungi in sclerophyll forest soil. [source]

    Detection of Heterobasidion annosum s. l. [(Fr.) Bref.] in Norway Spruce by Polymerase Chain Reaction

    JOURNAL OF PHYTOPATHOLOGY, Issue 7 2002
    G. Bahnweg
    Abstract Internal transcribed spacer (ITS) sequences of the rDNA repeat unit of Heterobasidion annosum were used to design specific primers for the detection and quantification of this important forest pathogen by polymerase chain reaction (PCR). Specificity of detection was cross-checked against a variety of other fungi (saprophytes, root pathogens, mycorrhizal fungi) which may occur in the same environment. As little as 1 pg fungal DNA (equiv. to 10,40 genomes) could be detected in 200 ng spruce root DNA (from 1 mg fresh spruce root). The Heterobasidion -specific primers allowed simultaneous detection of Armillaria spp. in multiplex PCR. The method was successfully applied to increment cores of Norway spruce from the forest region Tharandter Wald (Saxonia, Germany), Oberbärenburg (East Ore Mountains, Saxonia) and Oberschleissheim (north of Munich, Bavaria). [source]

    Genetic Identification of Pelagic Shark Body Parts for Conservation and Trade Monitoring

    CONSERVATION BIOLOGY, Issue 4 2002
    Mahmood Shivji
    Difficulties with the identification of many commonly fished sharks and their body parts has resulted in a global dearth of catch and trade information, making reliable assessment of exploitation effects and conservation needs for individual species nearly impossible. We developed and tested a highly streamlined molecular genetic approach based on species-specific, polymerase-chain-reaction primers in an eight-primer multiplex format to discriminate simultaneously between body parts from six shark species common in worldwide pelagic fisheries. The species-specific primers are based on DNA sequence differences among species in the nuclear ribosomal internal transcribed spacer 2 locus. The primers and multiplex format accurately and sensitively distinguished samples from each of three lamnid ( Isurus oxyrinchus, Isurus paucus, and Lamna nasus) and three carcharhinid ( Prionace glauca, Carcharhinus obscurus, and Carcharhinus falciformis) species from all but one other shark species encountered in the North Atlantic fishery. Furthermore, the three lamnid primers were robust enough in their discriminatory power to be useful for species diagnosis on a global scale. Preliminary testing of dried fins from Asian and Mediterranean commercial markets suggests that our genetic approach will be useful for determining the species of origin of detached fins, thus allowing the monitoring of trade in shark fins for conservation assessment. Our approach will also facilitate detection of products from protected and other at-risk shark species and may prove useful as a model for development of the high-throughput, genetic, species-diagnosis methods typically required in conservation and management contexts. Resumen: La conservación y manejo de tiburones fundamentado a nivel de especie es una necesidad imperativa debido a la creciente demanda de aletas de tiburón y el reconocimiento de que las especies individuales de tiburones responden de manera distinta a la explotación. Las dificultades para la identificación de muchos tiburones capturados comúnmente, así como de partes de su cuerpo, han resultado en una escasez global de información sobre capturas y comercialización, haciendo casi imposible el poder realizar evaluaciones de los efectos de la explotación y de las necesidades de conservación. Desarrollamos y evaluamos un método altamente estilizado de genética molecular basado en detonadores de la reacción en cadena de la polimerasa, especie-específicos, en un formato múltiple de ocho detonadores para discriminar simultáneamente entre las partes del cuerpo de seis especies de tiburones provenientes de pesquerías pelágicas mundiales comunes. Los detonadores especie-específicos están basados en diferencias en las secuencias de ADN entre especies del locus espaciador 2 nuclear, ribosomal, transcrito. Los detonadores y el formato múltiple distinguen muestras con precisión y sensitividad de cada uno de los tres lámnidos ( Isurus oxyrinchus, Isurus paucus y Lamna nasus) y tres especies de carcarínidos ( Prionace glauca, Carcharhinus obscurus y Carcharhinus falciformis) especies todas encontradas en las pesquerías de Norteamérica, excepto una. Mas aún, los detonadores de los tres lamnidos fueron lo suficientemente robustos en su poder discriminante como para ser usados para el diagnóstico de especies a escala mundial. Las pruebas preliminares de aletas secas de los mercados comerciales de Asia y el Mediterráneo sugieren que nuestro método genético puede ser útil para determinar la especie de origen de las aletas separadas, permitiendo así usar el monitoreo de las aletas de tiburón para evaluaciones de conservación. Nuestro método también podría facilitar la detección de productos provenientes de especies protegidas o en riesgo y podría resultar útil como un modelo para el desarrollo de métodos genéticos de alto rendimiento para el diagnóstico de especies, métodos típicamente requeridos en los contextos de conservación y manejo. [source]

    Range size, taxon age and hotspots of neoendemism in the California flora

    DIVERSITY AND DISTRIBUTIONS, Issue 3 2010
    Nathan J. B. Kraft
    Abstract Aim, Sustaining biological diversity requires the protection of the ecological, evolutionary and landscape-level processes that generate it. Here, we identify areas of high neoendemism in a global diversity hotspot, the California flora, using range size data and molecular-based estimates of taxon age. Location, California, USA. Methods, We compiled distribution and range size data for all plant taxa endemic to California and internal transcribed spacer (ITS)-based age estimates for 337 putative neoendemics (15% of the endemic flora). This information was combined to identify areas in the state with high proportions of young and restricted-range taxa. We overlaid the distribution of neoendemic hotspots on maps of currently protected lands and also explored correlations between our diversity measures and climate. Results, The central coast of California, the Sierra Nevada and the San Bernardino Range contained endemics with the most restricted distributions on average, while areas in the Desert and Great Basin provinces found within the state were composed of the youngest neoendemics on average. Diversity measures that took age and range size into account shifted the estimate of highest endemic diversity in the state towards the Desert and Great Basin regions relative to simple counts of endemic species richness. Our diversity measures were poorly correlated with climate and topographic heterogeneity. Main conclusions, Substantial portions of California with high levels of plant neoendemism fall outside of protected lands, indicating that additional action will be needed to preserve the geographic areas apparently associated with high rates of plant diversification. The neoendemic flora of the deserts appears particularly young in our analyses, which may reflect the relatively recent origin of desert environments within the state. [source]

    Naturally occurring egg parasitoids of the genus Trichogramma (Hymenoptera: Trichogrammatidae) in a pomegranate orchard in Tunisia

    ENTOMOLOGICAL SCIENCE, Issue 1 2010
    Ines KSENTINI
    Abstract Four Trichogramma species were found in a pomegranate orchard in Gabès, an arid region of Tunisia, from parasitized eggs of Ectomyelois ceratoniae Zeller (Lepidoptera: Pyralidae), an economically important insect pest. Identification based on assessment of male genitalia and internal transcribed spacer 2 (ITS2) sequences showed that they were T. bourarachae Pintureau and Babault, 1988, T. oleae Voegelé and Pointel, 1979, T. cacoeciae Marchal, 1927 and T. evanescens Westwood, 1833. Trichogramma evanescens is reported for the first time in Tunisia. Trichogramma cacoeciae was the largely dominant species in the analyzed samples, whereas T. bourarachae was present in a minor portion of 1.38%. The implications of these results for attempts at controlling E. ceratoniae are discussed. [source]

    Large-scale distribution and activity patterns of an extremely low-light-adapted population of green sulfur bacteria in the Black Sea

    ENVIRONMENTAL MICROBIOLOGY, Issue 5 2010
    Evelyn Marschall
    Summary The Black Sea chemocline represents the largest extant habitat of anoxygenic phototrophic bacteria and harbours a monospecific population of Chlorobium phylotype BS-1. High-sensitivity measurements of underwater irradiance and sulfide revealed that the optical properties of the overlying water column were similar across the Black Sea basin, whereas the vertical profiles of sulfide varied strongly between sampling sites and caused a dome-shaped three-dimensional distribution of the green sulfur bacteria. In the centres of the western and eastern basins the population of BS-1 reached upward to depths of 80 and 95 m, respectively, but were detected only at 145 m depth close to the shelf. Using highly concentrated chemocline samples from the centres of the western and eastern basins, the cells were found to be capable of anoxygenic photosynthesis under in situ light conditions and exhibited a photosynthesis,irradiance curve similar to low-light-adapted laboratory cultures of Chlorobium BS-1. Application of a highly specific RT-qPCR method which targets the internal transcribed spacer (ITS) region of the rrn operon of BS-1 demonstrated that only cells at the central station are physiologically active in contrast to those at the Black Sea periphery. Based on the detection of ITS-DNA sequences in the flocculent surface layer of deep-sea sediments across the Black Sea, the population of BS-1 has occupied the major part of the basin for the last decade. The continued presence of intact but non-growing BS-1 cells at the periphery of the Black Sea indicates that the cells can survive long-distant transport and exhibit unusually low maintenance energy requirements. According to laboratory measurements, Chlorobium BS-1 has a maintenance energy requirement of ,1.6,4.9·10,15 kJ cell,1 day,1 which is the lowest value determined for any bacterial culture so far. Chlorobium BS-1 thus is particularly well adapted to survival under the extreme low-light conditions of the Black Sea, and can be used as a laboratory model to elucidate general cellular mechanisms of long-term starvation survival. Because of its adaptation to extreme low-light marine environments, Chlorobium BS-1 also represents a suitable indicator for palaeoceanography studies of deep photic zone anoxia in ancient oceans. [source]

    Soils of a Mediterranean hot spot of biodiversity and endemism (Sardinia, Tyrrhenian Islands) are inhabited by pan-European, invasive species of Hypocrea/Trichoderma

    ENVIRONMENTAL MICROBIOLOGY, Issue 1 2009
    Quirico Migheli
    Summary We have used a Mediterranean hot spot of biodiversity (the Island of Sardinia) to investigate the impact of abiotic factors on the distribution of species of the common soil fungus Trichoderma. To this end, we isolated 482 strains of Hypocrea/Trichoderma from 15 soils comprising undisturbed and disturbed environments (forest, shrub lands and undisturbed or extensively grazed grass steppes respectively). Isolates were identified at the species level by the oligonucleotide BarCode for Hypocrea/Trichoderma (TrichOKEY), sequence similarity analysis (Trichoblast) and phylogenetic inferences. The majority of the isolates were positively identified as pan-European and/or pan-global Hypocrea/Trichoderma species from sections Trichoderma and Pachybasium, comprising H. lixii/T. harzianum, T. gamsii, T. spirale, T. velutinum, T. hamatum, H. koningii/T. koningii, H. virens/T. virens, T. tomentosum, H. semiorbis, H. viridescens/T. viridescens, H. atroviridis/T. atroviride, T. asperellum, H. koningiopsis/T. koningiopsis and Trichoderma sp. Vd2. Only one isolate represented a new, undescribed species belonging to the Harzianum,Catoptron Clade. Internal transcribed spacer sequence analysis revealed only one potentially endemic internal transcribed spacer 1 allele of T. hamatum. All other species exhibited genotypes that were already found in Eurasia or in other continents. Only few cases of correlation of species occurrence with abiotic factors were recorded. The data suggest a strong reduction of native Hypocrea/Trichoderma diversity, which was replaced by extensive invasion of species from Eurasia, Africa and the Pacific Basin. [source]

    Ecotype diversity in the marine picoeukaryote Ostreococcus (Chlorophyta, Prasinophyceae)

    ENVIRONMENTAL MICROBIOLOGY, Issue 6 2005
    Francisco Rodríguez
    Summary The importance of the cyanobacteria Prochlorococcus and Synechococcus in marine ecosystems in terms of abundance and primary production can be partially explained by ecotypic differentiation. Despite the dominance of eukaryotes within photosynthetic picoplankton in many areas a similar differentiation has never been evidenced for these organisms. Here we report distinct genetic [rDNA 18S and internal transcribed spacer (ITS) sequencing], karyotypic (pulsed-field gel electrophoresis), phenotypic (pigment composition) and physiological (light-limited growth rates) traits in 12 Ostreococcus strains (Prasinophyceae) isolated from various marine environments and depths, which suggest that the concept of ecotype could also be valid for eukaryotes. Internal transcribed spacer phylogeny grouped together four deep strains isolated between 90 m and 120 m depth from different geographical origins. Three deep strains displayed larger chromosomal bands, different chromosome hybridization patterns, and an additional chlorophyll (chl) c -like pigment. Furthermore, growth rates of deep strains show severe photo-inhibition at high light intensities, while surface strains do not grow at the lowest light intensities. These features strongly suggest distinct adaptation to environmental conditions encountered at surface and the bottom of the oceanic euphotic zone, reminiscent of that described in prokaryotes. [source]

    Application de la variabilité génétique de l'ADNr chez Monilinia laxa, Monilinia fructigena et Monilinia fructicolaà l'identification des espèces par PCR,

    EPPO BULLETIN, Issue 3-4 2000
    R. Ioos
    Monilinia laxa, Monilinia fructigena et Monilinia fructicola sont des agents de pourriture de fruit et de chancre sur rameau des arbres fruitiers. La région des ITS (internal transcribed spacer) située entre les gènes codant pour les sous-unités ribosomiques 18S et 28S de quatre isolats de M. fructigena et quatre isolats de M. fructigena collectés en France a été amplifiée par PCR et séquencée. L'alignement multiple de ces séquences d'ITS et leur comparaison avec les séquences d'ITS de Monilinia spp. publiées a révélé une très faible variabilité intraspécifique. Par contre, un faible polymorphisme interspécifique a été localisé au niveau de deux régions, respectivement dans l'ITS1 et l'ITS2. Ces deux régions ont été utilisées pour définir des couples d'oligonucléotides espèce-spécifiques. Ces couples d'amorces ont permis d'amplifier par PCR un fragment de 356 pb pour chacune des trois espèces. La spécificité des trois couples d'amorces a été vérifiée avec succès sur une collection de 17 isolats de M. laxa, 18 isolats de M. fructigena et 6 isolats de M. fructicola isolés de différents hôtes. En utilisant des conditions stringentes lors de la PCR, aucune réaction croisée n'a été observée avec les isolats testés. La spécificité du test a été d'autre part vérifiée avec l'ADN total extrait de différentes espèces de champignons, soit phylogénétiquement proche du genre Monilinia soit fréquemment isolées de fruits malades. L'utilisation de cette technique permet d'identifier avec fiabilité un isolaat indéterminé en une seule amplification, en le testant simultanément avec chacun des trois couples d'amorces. De plus, la détection et l'identification des espèces de Monilinia peuvent être réalisés directement à partir de fruits présentant des symptômes. Cette méthode simple et rapide pourrait être particulièrement utile pour détecter M. fructicola qui est un organisme de quarantaine pour l'Europe. [source]

    The use of ITS DNA sequence analysis and MALDI-TOF mass spectrometry in diagnosing an infection with Fusarium proliferatum

    EXPERIMENTAL DERMATOLOGY, Issue 11 2008
    Florian Seyfarth
    Abstract:, Although mycoses are among the most common diseases worldwide, infections with Fusarium spp. occur only rarely. Mostly patients suffering from underlying immune deficiency are infected with this mould, resulting in a considerably decreasing prognosis. In immunocompromised patients, cutaneous manifestations are more often associated with Fusarium sp. than with Candida sp. or Aspergillus sp. We describe one patient with acute lymphoblastic leukaemia, who was first treated with chemotherapy after GMALL protocol 07/03. After relapse, the patient was successfully transplanted in second remission with a human leukocyte antigen (HLA)-matched unrelated peripheral blood stem cell graft. Ten months later, the patient died from respiratory insufficiency and recurrence of leukaemia. Previously, Aspergillus antigen was detected in blood. In the latter course, disseminated papules appeared. One of these was examined histologically and mycologically. Conventional cultural diagnostics led to the diagnosis of a fusariosis, further supported by internal transcribed spacer (ITS) sequencing and matrix assisted laser desorption/ionisation,time-of-flight mass spectrometry (MALDI-TOF) mass spectrometry, both determining the isolated strain as Fusarium proliferatum, which is a very infrequent pathogen within this genus. Our investigations underline the potential of MALDI-TOF MS based identification of Fusarium species as an innovative, time and cost efficient alternative to ITS sequencing. [source]

    Phylogenetic diversity of Synechococcus strains isolated from the East China Sea and the East Sea

    FEMS MICROBIOLOGY ECOLOGY, Issue 3 2009
    Dong Han Choi
    Abstract Phylogenetic relationships among 33 Synechococcus strains isolated from the East China Sea (ECS) and the East Sea (ES) were studied based on 16S rRNA gene sequences and 16S,23S rRNA gene internal transcribed spacer (ITS) sequences. Pigment patterns of the culture strains were also examined. Based on 16S rRNA gene and ITS sequence phylogenies, the Synechococcus isolates were clustered into 10 clades, among which eight were previously identified and two were novel. Half of the culture strains belonged to clade V or VI. All strains that clustered into novel clades exhibited both phycoerythrobilin and phycourobilin. Interestingly, the pigment compositions of isolates belonging to clades V and VI differed from those reported for other oceanic regions. None of the isolates in clade V showed phycourobilin, whereas strains in clade VI exhibited both phycourobilin and phycoerythrobilin, which is in contrast to previous studies. The presence of novel lineages and the different pigment patterns in the ECS and the ES suggests the possibility that some Synechococcus lineages are distributed only in geographically restricted areas and have evolved in these regions. Therefore, further elucidation of the physiological, ecological, and genetic characteristics of the diverse Synechococcus strains is required to understand their spatial and geographical distribution. [source]

    Retrieval of first genome data for rice cluster I methanogens by a combination of cultivation and molecular techniques

    FEMS MICROBIOLOGY ECOLOGY, Issue 2 2005
    Christoph Erkel
    Abstract We report first insights into a representative genome of rice cluster I (RC-I), a major group of as-yet uncultured methanogens. The starting point of our study was the methanogenic consortium MRE50 that had been stably maintained for 3 years by consecutive transfers to fresh medium and anaerobic incubation at 50 °C. Process-oriented measurements provided evidence for hydrogenotrophic CO2 -reducing methanogenesis. Assessment of the diversity of consortium MRE50 suggested members of the families Thermoanaerobacteriaceae and Clostridiaceae to constitute the major bacterial component, while the archaeal population was represented entirely by RC-I. The RC-I population amounted to more than 50% of total cells, as concluded from fluorescence in situ hybridization using specific probes for either Bacteria or Archaea. The high enrichment status of RC-I prompted construction of a large insert fosmid library from consortium MRE50. Comparative sequence analysis of internal transcribed spacer (ITS) regions revealed that three different RC-I rrn operon variants were present in the fosmid library. Three, approximately 40-kb genomic fragments, each representative for one of the three different rrn operon variants, were recovered and sequenced. Computational analysis of the sequence data resulted in two major findings: (i) consortium MRE50 most likely harbours only a single RC-I genotype, which is characterized by multiple rrn operon copies; (ii) seven genes were identified to possess a strong phylogenetic signal (eIF2a, dnaG, priA, pcrA, gatD, gatE, and a gene encoding a putative RNA-binding protein). Trees exemplarily computed for the deduced amino acid sequences of eIF2a, dnaG, and priA corroborated a specific phylogenetic association of RC-I with the Methanosarcinales. [source]

    Potentiality of the cox1 gene in the taxonomic resolution of soil fungi

    FEMS MICROBIOLOGY LETTERS, Issue 1 2010
    Claire Molitor
    Abstract We explored the potential of the cox1 gene in the species resolution of soil fungi and compared it with the nuclear internal transcribed spacer (ITS) and small subunit (SSU)-rDNA. Conserved primers allowing the amplification of the fungal cox1 gene were designed, and a total of 47 isolates of Zygomycota and Ascomycota were investigated. The analysis revealed a lack of introns in >90% of the isolates. Comparison of the species of each of the six studied genera showed high interspecific sequence polymorphisms. Indeed, the average of nucleotide variations (4.2,11%) according to the genus, due mainly to the nucleotide substitutions, led to the taxonomic resolution of all the species studied regarding both ITS and SSU-rDNA, in which <88% were discriminated. The phylogenetic analysis performed after alignment of the cox1 gene across distant fungal species was in accordance with the well-known taxonomic position of the species studied and no overlap was observed between intra- and interspecific variations. These results clearly demonstrated that the cox1 sequences could provide good molecular markers for the determination of the species composition of environmental samples and constitute an important advance to study soil fungal biodiversity. [source]

    RESEARCH ARTICLE: Farysizyma gen. nov., an anamorphic genus in the Ustilaginales to accommodate three novel epiphytic basidiomycetous yeast species from America, Europe and Asia

    FEMS YEAST RESEARCH, Issue 3 2008
    João Inácio
    Abstract Among many isolates that resulted from four independent surveys of yeasts associated with plants in Brazil, the USA, Portugal and Taiwan, we have characterized eighteen basidiomycetous strains, two of which were conspecific with the type strain of Rhodotorula acheniorum, whereas the remaining sixteen isolates appeared not to correspond to any previously described species. Microsatellite-PCR fingerprinting with primers M13 and (GTG)5 confirmed that the latter strains formed three genetically distinct groups. Each group was considered to represent a distinct species based on nucleotide sequences of the D1/D2 domains of the 26S rRNA gene and the internal transcribed spacer (ITS) region. Phylogenetic analyses of sequence data placed the putative novel species in a clade with R. acheniorum and the dimorphic smut fungus Farysia chardoniana. A novel anamorphic genus, Farysizyma, is created to accommodate the three undescribed species, which were named Farysizyma itapuensis, Farysizyma setubalensis and Farysizyma taiwaniana. A new combination, Farysizyma acheniorum, is proposed for R. acheniorum, which may represent the yeast-phase anamorph of Farysia thuemenii. [source]

    The teleomorph of Chalara fraxinea, the causal agent of ash dieback

    FOREST PATHOLOGY, Issue 5 2009
    T. Kowalski
    Summary Ash dieback, caused by the pathogen Chalara fraxinea, is an emerging lethal disease of Fraxinus excelsior, threatening the host species in large parts of Europe. The ascomycete Hymenoscyphus albidus (Helotiaceae, Helotiales) was identified as the teleomorph of C. fraxinea by culturing from ascospores, morphological comparison and nuclear ribosomal internal transcribed spacer (ITS) sequencing. [source]

    PCR primers for identification of Sirococcus conigenus and S. tsugae, and detection of S. conigenus from symptomatic and asymptomatic red pine shoots

    FOREST PATHOLOGY, Issue 3 2008
    D. R. Smith
    Summary Regions of diversity in the internal transcribed spacer (ITS) sequences of Sirococcus species were exploited to design primer pairs used in a PCR-based method for the identification of the conifer shoot blight pathogen Sirococcus conigenus and the closely related fungus Sirococcus tsugae. The specificity of each primer pair for the respective fungus, detection limits and utility for detection from host material were confirmed. The S. conigenus primers were then used to detect this pathogen in tissues of symptomatic or apparently healthy red pine shoots collected at six locations in Wisconsin and Michigan and results compared with those obtained using a cultural assay. For needles, bark and wood of symptomatic shoots, the mean frequencies of detection of S. conigenus using the PCR-based methods were consistent (,7.5 out of 10) and always greater than for the cultural assay. Detection from symptomatic shoots using the cultural assay was more frequent from needles than from bark or wood. Both the PCR-based method and the cultural assay detected S. conigenus in similar frequencies from asymptomatic shoots, although less frequently than from symptomatic shoots. The efficiency of the PCR-based method and its utility for direct testing of host material should make it particularly useful in areas where multiple shoot blight pathogens are found. [source]

    Phylogeographic variation among isolates of the Sirococcus conigenus P group

    FOREST PATHOLOGY, Issue 1 2007
    H. Konrad
    Summary In this study the phylogeographic variation among isolates of the Sirococcus conigenus P group and the phylogenetic relationships of S. conigenus with Sirococcus clavigignenti-juglandacearum and other species previously placed in the genus Sirococcus were investigated. A collection of 33 isolates originating from Picea, Pinus and Larix in Europe, North America and Bhutan were characterized by sequence analyses of the internal transcribed spacer (ITS) region (including ITS1, 5.8S ribosomal DNA, ITS2) of the nuclear rDNA and a portion of the , -tubulin gene. In phylogenetic analyses most isolates from pine, spruce and larch formed a distinct clade, representing the P group of S. conigenus, which was separated from the T group of this pathogen. Four isolates from Picea in Europe and Canada formed a third clade within S. conigenus and these isolates are referred to as the S group. The P group consisted of five distinct ITS haplotypes, which partly differed in their optimum growth temperature and their growth rates at 25°C on malt extract agar. Nested clade analysis resolved the five haplotypes into three distinct clades and revealed significant genetic/geographic associations for some of the haplotypes. Parsimony analysis of the small subunit (18S) ribosomal DNA sequences confirmed the phylogenetic affinities between S. conigenus and S. clavigignenti-juglandacearum. In contrast, Godronia cassandrae and Hormococcus conorum, which formerly had been placed in the genus Sirococcus, were found to be only distantly related to S. conigenus and S. clavigignenti-juglandacearum. [source]

    Studies on anastomosis groups of Rhizoctonia solani isolates causing disease in two forest nurseries in Poland

    FOREST PATHOLOGY, Issue 2 2006
    S. St, pniewska-Jarosz
    Summary Thirty-eight isolates of Rhizoctonia spp. were isolated from Scots pine (Pinus sylvestris) seedlings with damping-off symptoms, originating from two forest nurseries in central-west Poland (Wronczyn and Jarocin) and from diseased seedlings grown in soil from Wronczyn nursery. Majority of these isolates (79%) had multinucleate cells and were identified as Rhizoctonia solani. The remaining isolates were recognized as binucleate Rhizoctonia spp. R. solani isolates were characterized using hyphal anastomosis and were divided into five anastomosis groups (AG). The most prevalent was AG5 (37% of isolates), followed by AG2-1 (30%) and 27% of the isolates were identified as AG4. Groups AG1-IB and AG2-2 were only represented by single isolates. The virulence recorded as mortality (in percentage) was comparatively high for binucleate and multinucleate isolates of Rhizoctonia spp. Sequence analysis of the polymerase chain reaction (PCR)-amplified internal transcribed spacer (ITS) rDNA region was used for phylogenetic analysis. The dendrogram showed that isolates were distinctly separated based on their AG types and there was no relationship between pathogenicity on Scots pine seedlings and the AG to which the isolates belong to. The results are discussed with respect to pathogenic potential of the various AG groups. Résumé Trente-huit isolats de Rhizoctonia spp. ont été isolés de semis de Pin sylvestre (Pinus sylvestris) présentant des symptômes de fonte, dans deux pépinières forestières du Centre-Ouest de la Pologne (Wronczyn and Jarocin) et de semis malades élevés dans du sol provenant de la pépinière de Wronczyn. La majorité de ces isolats (79%) ont des cellules multi-nucléées et ont été identifiés comme des Rhizoctonia solani. Le reste des isolats ont été reconnus comme des Rhizoctonia spp. binucléés. Les isolats de R. solani ont été caractérisés en utilisant l'anastomose d'hyphes et répartis dans cinq groupes d'anastomoses (AG). Le plus important est le groupe AG5 (37% des isolats), suivi par AG2-1 (30%) et AG4 (27%). Les groupes AG1-IB et AG2-2 sont représentés chacun par seulement un isolat. La virulence, estimée par le pourcentage de mortalité, est relativement forte pour les isolats binucléés et multinucléés de Rhizoctonia spp. L'analyse des séquences de la région ITS de l'ADNr amplifiées par PCR a été utilisée pour l'analyse phylogénétique. Le dendrogramme montre que les isolats sont séparés selon leur groupe d'anastomose mais il n'y a pas de relation entre le groupe d'anastomose et la virulence sur semis de Pin sylvestre. Les résultats sont discutés dans la perspective du pouvoir pathogène des différents groupes d'anastomoses. Zusammenfassung Von Kiefernsämlingen (Pinus sylvestris) mit Umfallkrankheit, die aus zwei Forstbaumschulen in Zentral-Westpolen stammten (Wronczyn und Jarocin) und aus erkrankten Sämlingen, die in Bodenproben aus der Baumschule Wronczyn kultiviert worden waren, wurden 38 Stämme von Rhizoctonia spp. isoliert. Die meisten dieser Isolate (79%) hatten vielkernige Zellen und wurden als R. solani identifiziert. Die restlichen Isolate waren zweikernige Rhizoctonia spp. Die Isolate von R. solani wurden durch Anastomosierungstests charakterisiert und fünf Anastomosierungsgruppen zugeordnet. Die häufigste Gruppe war AG5 (37% der Isolate), gefolgt von AG2-1 (30%) und AG4 (27%). Die Gruppe AG1-IB und AG2-2 waren nur durch einzelne Isolate vertreten. Die Virulenz (gemessen als % Mortalität) war sowohl für zweikernige als auch für vielkernige Isolate vergleichsweise hoch. Mit den Sequenzen der PCR-amplifizierten ITS-rDNA-Region wurde eine phylogenetische Analyse durchgeführt. Das Dendrogramm zeigte, dass die Isolate aufgrund ihrer Zugehörigkeit zu den Anastomosierungsgruppen deutlich voneinander getrennt waren, und es bestand keine Beziehung zwischen ihrer Virulenz gegenüber Kiefernsämlingen und der Gruppenzugehörigkeit. Die Befunde werden im Hinblick auf das pathogene Potential der verschiedenen Anastomosierungsgruppen diskutiert. [source]

    Rhizoctonia solani AG 2-1 as a causative agent of cotyledon rot on European beech (Fagus sylvatica)

    FOREST PATHOLOGY, Issue 6 2005
    A. M. Hietala
    Summary Rhizoctonia solani was frequently isolated in the Italian Alps from nursery-grown European beech (Fagus sylvatica) seedlings displaying symptoms of cotyledon rot. Koch's postulates were verified and mode of infection of the associated isolates was investigated with light and scanning electron microscopy. Population structure of the pathogen was investigated by scoring the anastomosis reaction type in pairings between different isolates from the same seedbed. One pathogen genotype showed a large distribution area within the seedbed, this implying that the inoculum had been building up in the seedbed over a longer time period. Hyphal anastomosis tests and sequence analysis of the internal transcribed spacer (ITS) region of ribosomal DNA indicated that the pathogen belongs to AG 2-1 of R. solani. ITS sequence analysis indicates that the isolates from beech are closely related to R. solani isolates causing a disease on tulip. The striking similarities between the two diseases are discussed. Résumé Rhizoctonia solani a fréquemment été isolé de semis de hêtre (Fagus sylvatica) présentant des symptômes de pourriture des cotylédons dans une pépinière forestière des Alpes italiennes. Les postulats de Koch ont été vérifiés et le mode d'infection étudié par microscopie optique et électronique à balayage. La structure de la population de l'agent pathogène a étéétudiée en examinant les réactions d'anastomoses dans les confrontations par paires des isolats d'un même lit de semences. Un génotype particulier s'est avéré largement distribué dans le lit de semence, suggérant soit une accumulation de l'inoculum pendant une longue période soit que ce génotype est capable de reproduction homocaryotique, favorisant sa dispersion. Les tests d'anastomose et l'analyse de la séquence de la région ITS de l'ADN ribosomal indiquent que l'agent pathogène appartient au groupe AG 2-1 de R. solani. L'analyse de la séquence de l'ITS montre que les isolats de hêtre sont proches d'isolats de R. solani pathogènes sur tulipe. Les ressemblances frappantes entre les deux maladies et la gestion de la maladie sur hêtre sont discutées. Zusammenfassung In einer Forstbaumschule in den italienischen Alpen wurde Rhizoctonia solani häufig aus Buchenkeimlingen (Fagus sylvatica) mit Symptomen einer Kotyledonenfäule isoliert. Die Koch'schen Postulate wurden erfüllt und die Art der Infektion der beteiligten Isolate wurde licht- und rasterelektronen-mikroskopisch untersucht. Die Populationsstruktur des Pathogens wurde anhand der Reaktionstypen (Anastomosierungsverhalten) in Paarungsversuchen mit den unterschiedlichen Isolaten aus demselben Saatbeet untersucht. Ein Kompatibilitätstyp war innerhalb des Saatbeetes weit verbreitet, was darauf hindeutet, dass sich das Inokulum über einen längeren Zeitraum dort angereichert hatte und/oder der Genotyp homokaryotisch fruchtet, was seine Ausbreitung fördert. Die Anastomosierungstests und die ITS-Sequenzanalyse der ribosomalen DNA ergaben, dass der Erreger zu der AG 2-1 Gruppe R. solani gehört. Die ITS-Sequenzen deuten darauf hin, dass die Isolate von Buche mit den R. solani, Isolaten verwandt sind, die an Tulpen pathogen sind. Die auffallende Ähnlichkeit der beiden Krankheiten und das Management der Erkrankung an Buche wird diskutiert. [source]

    Molecular diagnosis of Phytophthora lateralis in trees, water, and foliage baits using multiplex polymerase chain reaction

    FOREST PATHOLOGY, Issue 5 2001
    L. M. Winton
    A polymerase chain reaction (PCR)-based protocol for detection of Phytophthora lateralis in plant tissues and water is described. Base-pair (bp) deletions in both of the ribosomal DNA internal transcribed spacer (ITS) regions in P. lateralis were used to design complementary PCR primer sequences that amplify a 738 bp fragment only if P. lateralis DNA is present in the sample. Universal control primers based on conserved sequences of the nuclear ribosomal small subunit are included in a multiplexed reaction, providing an internal check on the procedure. The universal primers amplify an approximately 550 bp fragment that is common to plants, protists, and true fungi. The procedure reliably detects P. lateralis in cedar stem tissues and in roots. Positive reactions were obtained with as few as 200 P. lateralis zoospores in water. Diagnostic moléculaire par PCR multiplex pour détecter Phytophthora lateralis dans les arbres, l'eau et le feuillage utilisé comme piège Un protocole basé sur la PCR est décrit pour détecter Phytophthora lateralis dans les tissus végétaux et l'eau. Des délétions de paires de bases dans chacune des régions ITS de l'ADN ribosomal de P. lateralis ont été utilisées pour définir des amorces de PCR qui n'amplifient un fragment de 738 paires de bases que si l'ADN de P. lateralis est présent dans l'échantillon. Des amorces universelles basées sur des régions conservées de la petite sous-unité de l'ADN ribosomal nucléaire ont été incluses dans une réaction de PCR multiplex, fournissant ainsi un témoin interne de la réaction. Ces amorces universelles amplifient un fragment de 550 pb qui est commun aux plantes, aux protistes et aux champignons vrais. Ce protocole permet la détection de P. lateralis dans les tiges et dans les racines du Chamaecyparis. Des réactions positives ont été obtenues avec seulement 200 zoospores de P. lateralis dans l'eau. Molekulare Diagnose von Phytophthora lateralis in Bäumen, Wasser und als Köder benutzten Blättern mittels Multiplex-PCR Eine auf der PCR beruhende Methode zum Nachweis von Phytophthora lateralis in Pflanzengeweben und Wasser wird beschrieben. Deletionen in den beiden ITS Regionen der ribosomalen DNA von P. lateralis wurden zur Synthese von PCR-Primern ausgenutzt, die ein 738 Basenpaare langes Fragment nur dann amplifizieren, wenn P. lateralis in der Probe vorhanden ist. Universelle Primer, die konservierten Sequenzen der kleinen Unterheit der ribosomalen Kern-DNA entsprechen, wurden als interne Kontrollen in die Multiplex-PCR miteinbezogen. Diese Primer amplifizieren ein ungefähr 550 Basenpaare langes Fragment, das sowohl bei Pflanzen als auch bei Protisten und höheren Pilzen vorkommt. Mit der Methode liess sich P. lateralis im Stamm und in den Wurzeln von Lawsons Scheinzypresse verlässlich nachweisen. Für den Nachweis von P. lateralis im Wasser waren mindestens 200 Zoosporen nötig. [source]

    Molecular identification and phylogeny of East African Simulium damnosum s.l. and their relationship with West African species of the complex (Diptera: Simuliidae)

    INSECT MOLECULAR BIOLOGY, Issue 1 2000
    A. Krüger
    Abstract The phylogenetic relationships of East and West African members of the Simulium damnosum complex were studied by sequence analyses of the mitochondrial 16s ribosomal RNA (rRNA) gene. Results suggest that: (i) the S. damnosum complex is divided into an East and a West African clade, and (ii) S. pandanophilum and the cytoform ,Kiwira' form an East African subbranch distinct from the ,Sanje' group. In contrast to former assumptions from cytogenetic analyses, our molecular data do not support a direct relationship between the East African S. kilibanum and the West African S. squamosum. Length differences of the rDNA internal transcribed spacer 1 (ITS-1) turned out to be useful to distinguish between cytoforms. [source]

    Rapid and accurate identification of microorganisms contaminating cosmetic products based on DNA sequence homology

    INTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 6 2005
    Y. Fujita
    Synopsis The aim of this study was to develop rapid and accurate procedures to identify microorganisms contaminating cosmetic products, based on the identity of the nucleotide sequences of the internal transcribed spacer (ITS) region of the ribosomal RNA coding DNA (rDNA). Five types of microorganisms were isolated from the inner portion of lotion bottle caps, skin care lotions, and cleansing gels. The rDNA ITS region of microorganisms was amplified through the use of colony-direct PCR or ordinal PCR using DNA extracts as templates. The nucleotide sequences of the amplified DNA were determined and subjected to homology search of a publicly available DNA database. Thereby, we obtained DNA sequences possessing high similarity with the query sequences from the databases of all the five organisms analyzed. The traditional identification procedure requires expert skills, and a time period of approximately 1 month to identify the microorganisms. On the contrary, 3,7 days were sufficient to complete all the procedures employed in the current method, including isolation and cultivation of organisms, DNA sequencing, and the database homology search. Moreover, it was possible to develop the skills necessary to perform the molecular techniques required for the identification procedures within 1 week. Consequently, the current method is useful for rapid and accurate identification of microorganisms, contaminating cosmetics. Résumé Le but de cette étude est de développer une procédure rapide et fiable pour identifier les micro-organismes contaminant les produits cosmétiques. Cette procédure repose sur l'identification des séquences des nucléotides des espaceurs transcrits internes (Internal Transcribed Spacer ou région ITS), de l'ADN codant pour l'ARN ribosomique (rADN). Cinq types de micro-organismes sont isolés sur la partie intérieure des bouchons des flacons de lotions pour le soin de la peau et de gels lavants. Les régions ITS rADN des micro-organismes sont amplifiées grâce à l'utilisation de la méthode ,colony-direct PCR, ou ,ordinal PCR, en utilisant les extraits d'ADN comme matrices. Les séquences de nucléotides de l'ADN amplifiées sont évaluées et soumises à une recherche homologique dans une librairie d'ADN disponible au public. Ainsi, grâce aux bases de données, nous obtenons des séquences d'ADN qui possèdent une similaritéélevée avec les séquences recherchées des cinq organismes analysés. La procédure d'identification classique exige des compétences d'experts et une période d'environ un mois pour identifier les micro-organismes. D'autre part, il faut 3 à 7 jours pour terminer toutes les procédures utilisées dans la méthode ici décrite, y compris l'isolation et la culture des organismes, le séquençage de l'ADN et la recherche dermatologique dans les bases de données. De plus, il est possible en 1 semaine de développer les compétences nécessaires pour mettre en ,uvre les techniques moléculaires requises pour les procédures d'identification. Cette méthode est donc utile pour une identification rapide et fiable des micro-organismes qui contaminent les cosmétiques. [source]

    Surviving climate changes: high genetic diversity and transoceanic gene flow in two arctic,alpine lichens, Flavocetraria cucullata and F. nivalis (Parmeliaceae, Ascomycota)

    JOURNAL OF BIOGEOGRAPHY, Issue 8 2010
    József Geml
    Abstract Aim, We examined genetic structure and long-distance gene flow in two lichenized ascomycetes, Flavocetraria cucullata and Flavocetraria nivalis, which are widespread in arctic and alpine tundra. Location, Circumpolar North. Methods, DNA sequences were obtained for 90 specimens (49 for F. cucullata and 41 for F. nivalis) collected from various locations in Europe, Asia and North America. Sequences of the nuclear internal transcribed spacer (ITS) + 5.8S ribosomal subunit gene region were generated for 89 samples, and supplemented by beta-tubulin (BTUB) and translation elongation factor 1-alpha gene (EF1) sequences for a subset of F. cucullata specimens. Phylogenetic, nonparametric permutation methods and coalescent analyses were used to assess population divergence and to estimate the extent and direction of migration among continents. Results, Both F. cucullata and F. nivalis were monophyletic, supporting their morphology-based delimitation, and had high and moderately high intraspecific genetic diversity, respectively. Clades within each species contained specimens from both North America and Eurasia. We found only weak genetic differentiation among North American and Eurasian populations, and evidence for moderate to high transoceanic gene flow. Main conclusions, Our results suggest that both F. cucullata and F. nivalis have been able to migrate over large distances in response to climatic fluctuations. The high genetic diversity observed in the Arctic indicates long-term survival at high latitudes, whereas the estimated migration rates and weak geographic population structure suggest a continuing long-distance gene flow between continents that has prevented pronounced genetic differentiation. The mode of long-distance dispersal is unknown, but wind dispersal of conidia and/or ascospores is probably important in the open arctic landscapes. The high genetic diversity and efficient long-distance dispersal capability of F. cucullata and F. nivalis suggest that these species, and perhaps other arctic lichens as well, will be able to track their potential niche in the changing Arctic. [source]

    Incipient speciation of Catostylus mosaicus (Scyphozoa, Rhizostomeae, Catostylidae), comparative phylogeography and biogeography in south-east Australia

    JOURNAL OF BIOGEOGRAPHY, Issue 3 2005
    Michael N Dawson
    Abstract Aim, Phylogeography provides a framework to explain and integrate patterns of marine biodiversity at infra- and supra-specific levels. As originally expounded, the phylogeographic hypotheses are generalities that have limited discriminatory power; the goal of this study is to generate and test specific instances of the hypotheses, thereby better elucidating both local patterns of evolution and the conditions under which the generalities do or do not apply. Location, Coastal south-east Australia (New South Wales, Tasmania and Victoria), and south-west North America (California and Baja California). Methods, Phylogeographic hypotheses specific to coastal south-east Australia were generated a priori, principally from existing detailed distributional analyses of echinoderms and decapods. The hypotheses are tested using mitochondrial cytochrome c oxidase subunit I (COI) and nuclear internal transcribed spacer 1 (ITS1) DNA sequence data describing population variation in the jellyfish Catostylus mosaicus, integrated with comparable data from the literature. Results, Mitochondrial COI distinguished two reciprocally monophyletic clades of C. mosaicus (mean ± SD: 3.61 ± 0.40% pairwise sequence divergence) that were also differentiated by ITS1 haplotype frequency differences; the boundary between the clades was geographically proximate to a provincial zoogeographic boundary in the vicinity of Bass Strait. There was also limited evidence of another genetic inhomogeneity, of considerably smaller magnitude, in close proximity to a second hypothesized zoogeographic discontinuity near Sydney. Other coastal marine species also show genetic divergences in the vicinity of Bass Strait, although they are not closely concordant with each other or with reported biogeographic discontinuities in the region, being up to several hundreds of kilometres apart. None of the species studied to date show a strong phylogeographic discontinuity across the biogeographic transition zone near Sydney. Main conclusions, Patterns of evolution in the Bass Strait and coastal New South Wales regions differ fundamentally because of long-term differences in extrinsic factors. Since the late Pliocene, periods of cold climate and low sea-level segregated warm temperate organisms east or west of an emergent Bassian Isthmus resulting in population divergence and speciation; during subsequent periods of warmer and higher seas, sister taxa expanded into the Bass Strait region leading to weakly correlated phylogeographic and biogeographic patterns. The Sydney region, by contrast, has been more consistently favourable to shifts in species' ranges and long-distance movement, resulting in a lack of intra-specific and species-level diversification. Comparisons between the Sydney and Bass Strait regions and prior studies in North America suggest that vicariance plays a key role in generating coastal biodiversity and that dispersal explains many of the deviations from the phylogeographic hypotheses. [source]

    Evolution and biogeography of the austral genus Phyllocladus (Podocarpaceae)

    JOURNAL OF BIOGEOGRAPHY, Issue 10 2004
    Steven J. Wagstaff
    Abstract Aim, To infer evolutionary relationships within the genus Phyllocladus and among its close relatives by phylogenetic analysis of DNA sequences. Interpret the inferred relationships in association with the fossil record to examine the origin and diversification of the genus. Location, Australasia. Methods, Phylogenetic analyses of rbcL, matK and internal transcribed spacer (ITS) sequences representing all of the extant species of Phyllocladus and a selection of outgroups from Podocarpaceae and Araucariaceae. Results, The rbcL and matK sequences exhibit little variation within Phyllocladus, but ally its members to Podocarpaceae although its immediate sister remains unclear. The ITS sequences resolve all five species of Phyllocladus and two intraspecific ecotypes of P. alpinus. Main conclusions,Phyllocladus forms a distinct lineage that diverged early in the evolutionary history of Podocarpaceae. The fossil record indicates that the genus was more widely distributed and morphologically diverse during the early Tertiary than at present. Although of Mesozoic origin, the level of sequence variation within Phyllocladus suggests that the extant species radiated during the late Tertiary c. 6.3 ± 0.9 Ma. New Zealand is the present centre of species diversity. [source]

    Elevated genetic heterogeneity and Pleistocene climatic instability: inferences from nrDNA in New Zealand Coprosma (Rubiaceae)

    JOURNAL OF BIOGEOGRAPHY, Issue 7 2002
    Stephen R. Wichman
    Aim To examine patterns of hybridization and genotype mixing within the genus Coprosma J.R.Forst. & G.Forst. (Rubiaceae). Location New Zealand Methods Nucleotide sequence was determined for the internal transcribed spacer (ITS) and external transcribed spacer (ETS) regions of nuclear ribosomal DNA for fifty individuals from thirty-six taxa within the New Zealand component of the genus Coprosma. Results Mixed sequences were found to be widespread in Coprosma. Direct sequencing of ITS polymerase chain reaction (PCR) products from seven polyploid taxa showed evidence of sequence mixtures. Cloning and sequencing of individual PCR products from two polyploids confirmed the presence of multiple templates, one of which corresponded to that of a diploid. Intra-individual heterogeneity was also seen in a hybrid diploid taxon, with the mixed nucleotides corresponding to those of the parental lineages. Finally the ITS sequences of twenty-two diploid taxa showed that eleven contained intra-individual heterogeneity. Conclusions We conclude that the widespread occurrence of sequence mixtures in Coprosma results from of frequent hybridization. We also conclude that concerted evolution of the ITS and ETS regions is depressed. We propose that these characteristics evolved as a mechanism to maintain high levels of heterogeneity and suggest that this is adaptive for Coprosma in climatically unstable and physically complex New Zealand landscapes. These landscapes have been subjected to repeated oscillations between stadial and interstadial environments during the Pleistocene. [source]

    Evaluation of the alkaline wash/lysis procedure for the molecular diagnosis of a positive bacterial blood culture in clinical routine practice

    JOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 3 2010
    Sheng-Chuan Hsi
    Abstract Blood culture is commonly used to detect microorganisms in patients with a suspected blood infection. This study evaluated the alkaline wash/lysis procedure to extract DNA of microorganisms in a clinical blood culture. A multiplex polymerase chain reaction (PCR) targeting the 16S rDNA (ribosomal DNA) gene and the fungal ITS (internal transcribed spacer) gene was used as a reliable indicator for the presence of microorganism DNA in the extracts. A total of 535BacT/ALERT positive blood culture bottles were evaluated. Multiplex PCR showed positive results in 530 DNA extracts, but 5 DNA extracts gave negative results. We conclude that the alkaline wash/lysis procedure in combination with the multiplex PCR is a simple and sensitive method, which can be used in a standard diagnostic laboratory to detect microorganisms in blood culture material. J. Clin. Lab. Anal. 24:139,144, 2010. © 2010 Wiley-Liss, Inc. [source]

    Phylogenetic position of Salmo(Platysalmo)platycephalus Behnke 1968 from south-central Turkey, evidenced by genetic data

    JOURNAL OF FISH BIOLOGY, Issue 4 2004
    S. Su
    To determine whether the current classification of the flathead trout Salmo (Platysalmo) platycephalus, endemic to the upper reaches of the Zamanti River system, Turkey, based solely on morphology, is in congruence with molecular phylogeny, the nucleotide sequence variation in mitochondrial (control region and cytochrome b gene) and nuclear (internal transcribed spacer of rRNA genes) DNA for the flathead trout and various representatives of the genus Salmo was studied. On the basis of pair-wise genetic distance estimates, the highest differences were found to exist between the flathead trout and S. salar, S. ohridana and S. obtusirostris, whereas the differences between the flathead trout and S. trutta were minimal. All the analyses performed firmly positioned the flathead trout within the Adriatic phylogeographic lineage of S. trutta; however, the exact position of the flathead trout within the Adriatic cluster was irresolvable. Accordingly, classifying the flathead trout as a subgenus of Salmo is unjustifiable and its reclassification in a lower taxonomic category is suggested by the present study. [source]