A TaqMan real-time PCR assay for Rhizoctonia cerealis and its use in wheat and soil
Rhizoctonia cerealis causes sharp eyespot in cereals and the pathogen survives as mycelia or sclerotia in soil. Real-time Polymerase Chain Reaction (qPCR) assays based on TaqMan chemistry are highly suitable for use on DNA extracted from soil. We report here the first qPCR assay for R. cerealis usin...
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Springer
2017
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| Online Access: | https://eprints.nottingham.ac.uk/43567/ |
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| author | Woodhall, J.W. Brown, M.J. Perkins, K. Valdeolmillos, E. Somoza Boonham, N. Ray, Rumiana V. |
| author_facet | Woodhall, J.W. Brown, M.J. Perkins, K. Valdeolmillos, E. Somoza Boonham, N. Ray, Rumiana V. |
| author_sort | Woodhall, J.W. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Rhizoctonia cerealis causes sharp eyespot in cereals and the pathogen survives as mycelia or sclerotia in soil. Real-time Polymerase Chain Reaction (qPCR) assays based on TaqMan chemistry are highly suitable for use on DNA extracted from soil. We report here the first qPCR assay for R. cerealis using TaqMan primers and a probe based on a unique Sequence Characterised Amplified Region (SCAR). The assay is highly specific and did not amplify DNA from a range of other binucleate Rhizoctonia species or isolates of anastomosis groups of Rhizoctonia solani. The high sensitivity of the assay was demonstrated in soils using a bulk DNA extraction method where 200 μg sclerotia in 50 g of soil were detected. DNA of the pathogen could also be amplified from asymptomatic wheat plants. Using the assay on soil samples from fields under different crop rotations, R. cerealis was most frequently detected in soils where wheat was grown or soil under pasture. It was detected least frequently in fields where potatoes were grown. This study demonstrates that assays derived from SCAR sequences can produce specific and sensitive qPCR assays. |
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| format | Article |
| id | nottingham-43567 |
| institution | University of Nottingham Malaysia Campus |
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| last_indexed | 2025-11-14T19:52:24Z |
| publishDate | 2017 |
| publisher | Springer |
| recordtype | eprints |
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| spelling | nottingham-435672020-05-04T19:57:16Z https://eprints.nottingham.ac.uk/43567/ A TaqMan real-time PCR assay for Rhizoctonia cerealis and its use in wheat and soil Woodhall, J.W. Brown, M.J. Perkins, K. Valdeolmillos, E. Somoza Boonham, N. Ray, Rumiana V. Rhizoctonia cerealis causes sharp eyespot in cereals and the pathogen survives as mycelia or sclerotia in soil. Real-time Polymerase Chain Reaction (qPCR) assays based on TaqMan chemistry are highly suitable for use on DNA extracted from soil. We report here the first qPCR assay for R. cerealis using TaqMan primers and a probe based on a unique Sequence Characterised Amplified Region (SCAR). The assay is highly specific and did not amplify DNA from a range of other binucleate Rhizoctonia species or isolates of anastomosis groups of Rhizoctonia solani. The high sensitivity of the assay was demonstrated in soils using a bulk DNA extraction method where 200 μg sclerotia in 50 g of soil were detected. DNA of the pathogen could also be amplified from asymptomatic wheat plants. Using the assay on soil samples from fields under different crop rotations, R. cerealis was most frequently detected in soils where wheat was grown or soil under pasture. It was detected least frequently in fields where potatoes were grown. This study demonstrates that assays derived from SCAR sequences can produce specific and sensitive qPCR assays. Springer 2017-06 Article PeerReviewed Woodhall, J.W., Brown, M.J., Perkins, K., Valdeolmillos, E. Somoza, Boonham, N. and Ray, Rumiana V. (2017) A TaqMan real-time PCR assay for Rhizoctonia cerealis and its use in wheat and soil. European Journal of Plant Pathology, 148 (2). pp. 237-245. ISSN 1573-8469 Soil DNA; Crop rotation; Anastomosis group (AG); Sharp eyespot https://link.springer.com/article/10.1007%2Fs10658-016-1083-7 doi:10.1007/s10658-016-1083-7 doi:10.1007/s10658-016-1083-7 |
| spellingShingle | Soil DNA; Crop rotation; Anastomosis group (AG); Sharp eyespot Woodhall, J.W. Brown, M.J. Perkins, K. Valdeolmillos, E. Somoza Boonham, N. Ray, Rumiana V. A TaqMan real-time PCR assay for Rhizoctonia cerealis and its use in wheat and soil |
| title | A TaqMan real-time PCR assay for Rhizoctonia cerealis and its use in wheat and soil |
| title_full | A TaqMan real-time PCR assay for Rhizoctonia cerealis and its use in wheat and soil |
| title_fullStr | A TaqMan real-time PCR assay for Rhizoctonia cerealis and its use in wheat and soil |
| title_full_unstemmed | A TaqMan real-time PCR assay for Rhizoctonia cerealis and its use in wheat and soil |
| title_short | A TaqMan real-time PCR assay for Rhizoctonia cerealis and its use in wheat and soil |
| title_sort | taqman real-time pcr assay for rhizoctonia cerealis and its use in wheat and soil |
| topic | Soil DNA; Crop rotation; Anastomosis group (AG); Sharp eyespot |
| url | https://eprints.nottingham.ac.uk/43567/ https://eprints.nottingham.ac.uk/43567/ https://eprints.nottingham.ac.uk/43567/ |