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Fungal DNA (fungal + dna)

Distribution by Scientific Domains

Medical Sciences	50%
Life Sciences	50%

Selected Abstracts

Fungal rDNA signatures in coronary atherosclerotic plaques

ENVIRONMENTAL MICROBIOLOGY, Issue 12 2007
Stephan J. Ott
Summary Bacterial DNA has been found in coronary plaques and it has therefore been concluded that bacteria may play a role as trigger factors in the chronic inflammatory process underlying coronary atherosclerosis. However, the microbial spectrum is complex and it is not known whether microorganisms other than bacteria are involved in coronary disease. Fungal 18S rDNA signatures were systematically investigated in atherosclerotic tissue obtained through catheter-based atherectomy of 38 patients and controls (unaffected coronary arteries) using clone libraries, denaturating gradient gel analysis (DGGE), in situ hybridization and fluorescence in situ hybridization (FISH). Fungal DNA was found in 35 of 38 (92.11%) coronary heart disease patients by either polymerase chain reaction (PCR) with universal primers or in situ hybridization analysis (n = 5), but not in any control sample. In a clone library with more than 350 sequenced clones from pooled patient DNA, an overall richness of 19 different fungal phylotypes could be observed. Fungal profiles of coronary heart disease patients obtained by DGGE analysis showed a median richness of fungal species of 5 (range from 2 to 9) with a high interindividual variability (mean similarity 18.83%). For the first time, the presence of fungal components in atherosclerotic plaques has been demonstrated. Coronary atheromatous plaques harbour diverse and variable fungal communities suggesting a polymicrobial contribution to the chronic inflammatory aetiology. [source]

Analysing scots pine defence-related transcripts and fungal DNA levels in seedlings single- or dual-inoculated with endophytic and pathogenic Rhizoctonia species

FOREST PATHOLOGY, Issue 6 2009
H. Grönberg
Summary Fungal DNA and induction of host defence-related transcripts were monitored by real-time PCR in young Scots pine seedlings inoculated with pathogenic uninucleate (UNR) and endophytic binucleate (BNR) Rhizoctonia species. The UNR (teleomorph Ceratobasidium bicorne) causes root dieback in conifer seedlings following invasion of the vascular cylinder via root apex and destroying apical meristems whilst the BNR, representing anastomosis group AG-I of genus Ceratobasidium, is primarily restricted to the cortex in basal root regions. In the experiment 1 the fungi were simultaneously inoculated on roots, while in experiment 2, BNR was pre-inoculated 168 h before inoculation with UNR. Nucleic acids were extracted from infected roots at intervals up to 192 h post-infection (hpi), and the genomic DNA levels of the host and fungi and the transcript levels of a house-keeping gene (glyceraldehyde-3-phosphate dehydrogenase) and nine putative defence genes were quantified. In simultaneous inoculation UNR was more competitive than BNR whereas pre-inoculation of BNR suppressed but did not completely prevent root colonization by UNR. Stilbene synthase (STS) transcription was significantly up-regulated in single-inoculations with both fungi and in dual inoculation in both experiments. Maximum STS transcript levels were observed in roots single-inoculated with UNR; the peak level at 48 hpi in experiment 2 was significantly higher than in seedlings single-inoculated with BNR or co-inoculated with both fungi, the latter two treatments showing relatively similar STS transcript levels. Similarly, transcript levels of phenylalanine ammonia lyase at 48 hpi in experiment 2 were significantly higher in roots single-inoculated with UNR compared with BNR or in UNR+BNR co-inoculations. The other seven putative defence genes monitored did not show any clear-cut up-regulation following fungal inoculation. We conclude that BNR suppresses UNR in Scots pine roots via direct competition for infection sites, since the studied transcripts showed no evidence of BNR induced resistance against UNR. [source]

Seasonal and substrate preferences of fungi colonizing leaves in streams: traditional versus molecular evidence

ENVIRONMENTAL MICROBIOLOGY, Issue 2 2005
Liliya G. Nikolcheva
Summary Aquatic hyphomycetes are the main fungal decomposers of plant litter in streams. We compared the importance of substrate (three leaf species, wood) and season on fungal colonization. Substrates were exposed for 12 4-week periods. After recovery, mass loss, fungal biomass and release of conidia by aquatic hyphomycetes were measured. Fungal communities were characterized by counting and identifying released conidia and by extracting and amplifying fungal DNA (ITS2), which was subdivided into phylotypes by denaturing gradient gel electrophoresis (DGGE) and terminal-restriction fragment length polymorphism (T-RFLP). Mass loss, fungal biomass and reproduction were positively correlated with stream temperature. Conidial diversity was highest between May and September. Numbers of different phylotypes were more stable. Principal coordinate analyses (PCO) and canonical analyses of principal coordinates (CAP) of presence/absence data (DGGE bands, T-RFLP peaks and conidial species) showed a clear seasonal trend (P, 0.002) but no substrate effect (P, 0.88). Season was also a significant factor when proportional similarities of conidial communities or relative intensities of DGGE bands were evaluated (P, 0.003). Substrate was a significant factor determining DGGE band intensities (P = 0.002), but did not significantly affect conidial communities (P = 0.50). Both traditional and molecular techniques suggest that strict exclusion of fungi by substrate type is rare, and that presence of different species or phylotypes is governed by season. Biomasses of the various taxa (based on DGGE band intensities) were related to substrate type. [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]

Use of magnetic beads to extract fungal DNA

MYCOSES, Issue 1 2005
E. Faggi
Summary Authors compare two methods of extracting DNA from different fungi: the classic method with phenol/chloroform (P/C) and that with magnetic beads. Both were tested on Candida albicans and Cryptococcus neoformans var. neoformans, belonging to the yeast group and Microsporum canis, M. gypseum, Trichophyton rubrum, T. interdigitale, T. ajelloi, Epidermophyton floccosum, belonging to the dermatophytes group. Extraction products underwent polymerase chain reaction (PCR) fingerprinting with the appropriate primers to point out any disagreement in the genomic profiles. After having determined that the genomic profiles obtained from the DNA extracted from the same strain with the two methods correspond perfectly, the authors concluded that the extraction method with magnetic beads from fungal cells is simpler and quicker than with P/C extraction, greatly facilitating the obtainment of fungal DNA. [source]

Molecular detection of dermatophytes and nondermatophytes in onychomycosis by nested polymerase chain reaction based on 28S ribosomal RNA gene sequences

BRITISH JOURNAL OF DERMATOLOGY, Issue 5 2009
M. Ebihara
Summary Background, Onychomycosis is often caused by dermatophytes, but the role of nondermatophytes is underestimated due to the difficulty of identifying them by conventional direct microscopy and culture. Objectives, This study aims to detect nondermatophytes, as well as dermatophytes, in the nail samples of patients with onychomycosis using a polymerase chain reaction (PCR)-based culture-independent method. Materials and methods, The nested PCR assay targeting the sequence of the 28S ribosomal RNA gene was used to amplify fungal DNAs from 50 microscopy-positive nail specimens. Newly designed primer sets for dermatophyte universal, Trichophyton rubrum, T. mentagrophytes, Aspergillus spp., Scopulariopsis brevicaulis, Fusarium solani, F. oxysporum, F. verticillioides, Candida albicans and C. tropicalis were used after confirmation of their specificity. Results, Forty-seven cases (94%) were positive for fungal DNA, among which dermatophytes were detected in 39 cases (83·0%): T. rubrum in 35 cases (74·5%) and T. mentagrophytes in eight cases (17·0%). Surprisingly, nondermatophytes were detected in 18 cases (38·3%), both dermatophytes and nondermatophytes in 10 cases (21·3%) and nondermatophytes alone in eight cases (17·0%). Aspergillus spp. alone was observed in five cases (10·6%). Conclusions, This study indicates that most of the affected nail plates of patients with onychomycosis were positive for specific fungal DNAs, and suggests that nondermatophytes detected at high rates may be involved in the pathogenesis of onychomycosis. [source]

Culturing and direct DNA extraction find different fungi from the same ericoid mycorrhizal roots

NEW PHYTOLOGIST, Issue 1 2003
Tamara R. Allen
Summary ,,This study compares DNA and culture-based detection of fungi from 15 ericoid mycorrhizal roots of salal (Gaultheria shallon), from Vancouver Island, BC Canada. ,,From the 15 roots, we PCR amplified fungal DNAs and analyzed 156 clones that included the internal transcribed spacer two (ITS2). From 150 different subsections of the same roots, we cultured fungi and analyzed their ITS2 DNAs by RFLP patterns or sequencing. We mapped the original position of each root section and recorded fungi detected in each. ,,Phylogenetically, most cloned DNAs clustered among Sebacina spp. (Sebacinaceae, Basidiomycota). Capronia sp. and Hymenoscyphus erica (Ascomycota) predominated among the cultured fungi and formed intracellular hyphal coils in resynthesis experiments with salal. ,,We illustrate patterns of fungal diversity at the scale of individual roots and compare cloned and cultured fungi from each root. Indicating a systematic culturing detection bias, Sebacina DNAs predominated in 10 of the 15 roots yet Sebacina spp. never grew from cultures from the same roots or from among the > 200 ericoid mycorrhizal fungi previously cultured from different roots from the same site. [source]

Molecular detection of dermatophytes and nondermatophytes in onychomycosis by nested polymerase chain reaction based on 28S ribosomal RNA gene sequences