Rapid nested PCR-based detection of Ramularia collo-cygni direct from barley
Open Access
- 1 March 2006
- journal article
- research article
- Published by Oxford University Press (OUP) in FEMS Microbiology Letters
- Vol. 256 (2), 217-223
- https://doi.org/10.1111/j.1574-6968.2006.00121.x
Abstract
Ramularia collo-cygni is a barley pathogen of increasing importance in Northern and Central Europe, New Zealand and South America. Accurate visual and microscopic identification of the pathogen from diseased tissue is difficult. A nested PCR-based diagnostic test has been developed as part of an initiative to map the distribution of the pathogen in Scotland. The entire nuclear ribosomal internal transcribed spacer and 5.8S rRNA gene regions from 14 isolates of diverse global origin exhibited complete homology following sequence characterization. Two pairs of species-specific primers, based on inter-specific sequence divergence with closely related species, were designed and empirically evaluated for diagnostic nested PCR. Nested primers Rcc3 and Rcc4 consistently amplified a single product of 256 bp from DNA of 24 R. collo-cygni isolates of diverse global provenance, but not from other Ramularia species, or other fungi commonly encountered in cereal pathosystems, as well as Hordeum or Secale DNA preparations. Using this approach, R. collo-cygni was successfully identified from naturally infected barley leaf, awn and grain samples of diverse geographical provenance, in particular from symptoms that lacked the presence of characteristic conidiophores. It is envisaged that this assay will become established as an important tool in continuing studies into the ecology, aetiology and epidemiology of this poorly understood yet economically damaging plant pathogen.Keywords
This publication has 11 references indexed in Scilit:
- Development of a nested PCR detection procedure forNectria fuckelianadirect from Norway spruce bark extractsFEMS Microbiology Letters, 2005
- Quantitative analysis of phytopathogenic ascomycota on leaves of pedunculate oaks (Quercus roburL.) by real-time PCRFEMS Microbiology Letters, 2002
- Molecular detection of Neonectria galligena (syn. Nectria galligena)Mycological Research, 2002
- A Phylogenetic Redefinition of Anamorph Genera in Mycosphaerella Based on ITS rDNA Sequence and MorphologyMycologia, 2001
- A PCR-Based Assay to Detect Rhynchosporium secalis in Barley SeedPlant Disease, 2001
- PCR detection of Gremmeniella abietina, the causal agent of Scleroderris canker of pineMycological Research, 2000
- Rapid detection and diagnosis ofSeptoria triticiepidemics in wheat using a polymerase chain reaction/PicoGreen assayJournal of Applied Microbiology, 1999
- Differentiation of Gaeumannomyces graminis from other turf‐grass fungi by amplification with primers from ribosomal internal transcribed spacersPlant Pathology, 1995
- CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choiceNucleic Acids Research, 1994
- Taxonomy of Ophiocladium hordei, causing leaf lesions on Triticale and other GramineaeTransactions of the British Mycological Society, 1988