Detection of Pasteuria penetrans infection in Meloidogyne arenaria race 1 in planta by polymerase chain reaction
| Autor: | Guang Nong, James F. Preston, D. W. Dickson, Liesbeth M. Schmidt, H. C. Aldrich |
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| Rok vydání: | 2009 |
| Předmět: |
DNA
Bacterial Sequence Homology Gram-Positive Bacteria Applied Microbiology and Biotechnology Microbiology DNA Ribosomal Polymerase Chain Reaction law.invention Bacterial Proteins Solanum lycopersicum law RNA Ribosomal 16S medicine Animals Cluster Analysis Tylenchoidea Gene Ribosomal DNA Polymerase chain reaction Gram-Positive Bacterial Infections Phylogeny DNA Primers Ecology biology medicine.diagnostic_test fungi biology.organism_classification 16S ribosomal RNA Pasteuria RNA Bacterial Meloidogyne arenaria Primer (molecular biology) Fluorescence in situ hybridization Bacillus subtilis |
| Zdroj: | FEMS microbiology ecology. 48(3) |
| ISSN: | 1574-6941 |
| Popis: | We report on the development of a PCR-based assay to detect Pasteuria penetrans infection of Meloidogyne arenaria in planta using specific primers for recently sequenced sigE, spoIIAB and atpF genes of P. penetrans biotype P20. Amplification of these genes in crude DNA extracts of ground tomato root galls using real-time kinetic PCR distinguished infected from uninfected M. arenaria race 1 by analysis of consensus thresholds for single copy genes. Fluorescent in situ hybridization (FISH) using the sigE primer sequence as a probe shows hybridization to P. penetrans cells in various stages of vegetative (pre-endospore) development. Ratios of gene copies for sigE and 16S rDNA were obtained for P. penetrans and compared to Bacillus subtilis as a genomic paradigm of endospore-forming bacteria. Phylogenetic analysis of the sigE gene from Gram-positive, endospore-forming bacteria finds P. penetrans most closely related Paenbacillus polymyxa. The sporulation genes (spo genes), particularly sigE, have sequence diversity that recommends them for species and biotype differentiation of the numerous Pasteuria isolates that infect a large number of plant-parasitic nematodes. |
| Databáze: | OpenAIRE |
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