Rapid PCR-based method for detection and differentiation of Didymiaceae and Physaraceae (myxomycetes) in environmental samples
| Autor: | Akiko Kamono, Manabu Fukui |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Microbiology (medical)
Sequence analysis Molecular Sequence Data Biology Microbiology DNA Ribosomal Polymerase Chain Reaction Environmental Microbiology RNA Ribosomal 18S Animals Myxomycetes Molecular Biology Soil Microbiology DNA Primers Genetics Polymorphism Genetic Intron RNA Sequence Analysis DNA Ribosomal RNA Amplicon DNA Protozoan Molecular biology Introns Primer (molecular biology) Soil microbiology Temperature gradient gel electrophoresis |
| Zdroj: | Journal of microbiological methods. 67(3) |
| ISSN: | 0167-7012 |
| Popis: | Ecological studies of myxomycetes have been limited by the absence of universal cultivation techniques and the lack of life stage independent identification methods. We designed a novel PCR primer pair for the specific amplification of small subunit ribosomal RNA gene of Didymiaceae and Physaraceae. The primers produced amplicons from 192 fruiting body samples belonging to 10 genera. Twenty-four samples yielded longer fragments and sequence analysis revealed the presence of intron(s). As for the exonic regions, while sequence heterogeneities within a single species/varietas/forma were frequently observed, identical sequences were obtained only from identical species/varietas. The effectiveness of this primer pair in the analysis of morphologically unidentifiable samples was confirmed with the applications to samples of environmental plasmodium/sclerotium and soil. Denaturing gradient gel electrophoresis analysis was also tested with the soil samples. The results presented here demonstrate this PCR-based method can facilitate further ecological studies of Physaraceae and Didymiaceae in the environment. |
| Databáze: | OpenAIRE |
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