European Journal of Plant Pathology

, Volume 136, Issue 2, pp 273–280 | Cite as

A new quantitative real-time PCR assay for Rhizoctonia solani AG3-PT and the detection of AGs of Rhizoctonia solani associated with potato in soil and tuber samples in Great Britain

  • James W. Woodhall
  • Ian P. Adams
  • Jeff C. Peters
  • Glyn Harper
  • Neil Boonham


Rhizoctonia solani is an important pathogen of potatoes causing stem canker and black scurf. The fungus is a species complex comprised of 13 known anastomosis groups (AGs). AG3-PT is the anastomosis group frequently associated with disease in potatoes. A real-time PCR assay was designed to the rDNA ITS region of AG3-PT isolates to enable the pathogen to be detected directly in tuber and soil samples. The resulting assay was highly specific for AG3-PT, and did not amplify DNA from isolates from other AGs or subgroups of AG3. Using a bulk DNA extraction method capable of extracting from up to 250 g of soil, the assay could detect one individual sclerotium of AG3-PT (weighing 200 μg) in 250 g of soil. The AG3-PT assay was used, with assays for AG2-1, AG5 and AG8 to determine the prevalence of those AGs in UK potato soils and tubers. AG2-1 and AG3-PT were the predominant groups in tubers and soils, although AG3-PT was more frequently isolated from tubers, highlighting its importance as a potato pathogen. AG3-PT was also detected in more than half of the tuber samples tested suggesting the importance of seed borne inoculum.


Rhizoctonia solani AG3-PT Potatoes Real-time PCR Soil 



Anastomosis group


Cycle threshold


Internal transcribed spacer


Potato dextrose agar


Ribosomal DNA


  1. Balali, G. R., Neate, S. M., Scott, E. S., Whisson, D. L., & Wicks, T. J. (1995). Anastomosis group and pathogenicity of isolates of Rhizoctonia solani from potato crops in South Australia. Plant Pathology, 44, 1050–1057.CrossRefGoogle Scholar
  2. Bandy, B. P., Leach, S. S., & Tavantzis, S. M. (1988). AG3 is the major cause of rhizoctonia disease of potato in Maine. Plant Disease, 72, 596–598.CrossRefGoogle Scholar
  3. Banville, G. J., Carling, D. E., & Otrysko, B. E. (1996). Rhizoctonia disease on potato. In B. Sneh, S. Jabaji-Hare, S. Neate, & G. Dijst (Eds.), Rhizoctonia species. Taxonomy, molecular biology, ecology, pathology and disease control (pp. 321–330). Dordrecht: Kluwer.Google Scholar
  4. Bartz, F. E., Cubeta, M. A., Toda, T., Naito, S., & Ivors, K. L. (2010). An in planta method for assessing the role of basidiospores in Rhizoctonia foliar disease of tomato. Plant Disease, 94, 515–520.CrossRefGoogle Scholar
  5. Boonham, N., Glover, R., Tomlinson, J., & Mumford, R. (2008). Exploiting generic platform technologies for the detection and identification of plant pathogens. European Journal of Plant Pathology, 121, 355–363.CrossRefGoogle Scholar
  6. Brierley, J. L., Stewart, J. A., & Lees, A. K. (2009). Quantifying potato pathogen DNA in soil. Applied Soil Ecology, 41, 234–238.CrossRefGoogle Scholar
  7. Budge, G. E., Shaw, M. W., Colyer, A., Pietravalle, S., & Boonham, N. (2009a). Molecular tools to investigate Rhizoctonia solani distribution in soil. Plant Pathology, 58, 1071–1080.CrossRefGoogle Scholar
  8. Budge, G. E., Shaw, M. W., Lambourne, C., Jennings, P., Clayburn, R., Boonham, N., & McPherson, M. (2009b). Characterization and origin of infection of Rhizoctonia solani associated with Brassica oleracea crops in the UK. Plant Pathology, 58, 1059–1070.CrossRefGoogle Scholar
  9. Campion, C., Chatot, C., Perraton, B., & Andrivon, D. (2003). Anastomosis groups, pathogenicity and sensitivity to fungicides of Rhizoctonia solani isolates collected on potato crops in France. European Journal of Plant Pathology, 109, 983–992.CrossRefGoogle Scholar
  10. Carling, D. E., & Leiner, R. H. (1986). Isolation and characterisation of Rhizoctonia solani and binucleate Rhizoctonia solani-like fungi from aerial stems and subterranean organs of potato plants. Phytopathology, 76, 725–729.CrossRefGoogle Scholar
  11. Ceresini, P. C., Shew, H. D., Vilgalys, R. J., & Cubeta, M. A. (2002). Genetic diversity of Rhizoctonia solani AG3 from potato and tobacco in North Carolina. Mycologia, 94, 437–449.PubMedCrossRefGoogle Scholar
  12. Chand, T., & Logan, C. (1983). Cultural and pathogenic variation in potato isolates of Rhizoctonia solani in Northern Ireland. Transactions of the British Mycological Society, 81, 585–589.CrossRefGoogle Scholar
  13. Fiers, M., Chatot, C., Edel-Hermann, V., Le Hingrat, Y., Yanougo, A. K., Gautheron, N., Guillery, E., Alabouvette, C., & Steinberg, C. (2010). Diversity of microorganisms associated with atypical superficial blemishes of potato tubers and pathogenicity assessment. European Journal of Plant Pathology, 128, 353–37.CrossRefGoogle Scholar
  14. Hide, G. A., & Firmager, J. P. (1990). Effects of an isolate of Rhizoctonia solani Kühn AG8 from diseased barley on the growth and infection of potatoes (Solanum tuberosum). Potato Research, 33, 229–234.CrossRefGoogle Scholar
  15. Lees, A. K., Cullen, D. W., Sullivan, L., & Nicolson, M. J. (2002). Development of conventional and quantitative real-time PCR assays for the detection and identification of Rhizoctonia solani AG3 in potato and soil. Plant Pathology, 51, 293–302.CrossRefGoogle Scholar
  16. Lehtonen, M. J., Ahvenniemi, P., Wilson, P. S., German-Kinnari, M., & Valkonen, J. P. T. (2008). Biological diversity of Rhizoctonia solani (AG-3) in a northern potato-cultivation environment in Finland. Plant Pathology, 57, 141–151.Google Scholar
  17. Ophel-Keller, K., Mckay, A., Hartley, D., Herdina, & Curran, J. (2008). Development of a routine DNA-based testing service for soilborne diseases in Australia. Australasian Plant Pathology, 37, 243–253.CrossRefGoogle Scholar
  18. Sneh, B., Burpee, L., & Ogoshi, A. (1991). Identification of Rhizoctonia species. St. Paul: APS Press.Google Scholar
  19. Tsror, L. (2010). Biology, epidemiology and management of Rhizoctonia solani on potato. Journal of Phytopathology, 158, 649–658.CrossRefGoogle Scholar
  20. Woodhall, J. W., Lees, A. K., Edwards, S. G., & Jenkinson, P. (2007). Characterization of Rhizoctonia solani from potato in Great Britain. Plant Pathology, 56, 286–295.CrossRefGoogle Scholar
  21. Woodhall, J. W., Lees, A. K., Edwards, S. G., & Jenkinson, P. (2008). Infection of potato by Rhizoctonia solani: effect of anastomosis group. Plant Pathology, 57, 897–905.CrossRefGoogle Scholar
  22. Woodhall, J. W., Webb, K. M., Harper, G., Peters, J. C., Rodriguez-Carres, M., & Cubeta, M. A. (2011). First report of a new binucleate Rhizoctonia on potato tubers in the UK. New Disease Reports, 23, 31. doi:10.5197/j.2044-0588.2011.023.031.CrossRefGoogle Scholar
  23. Woodhall, J. W., Webb, K. M., Giltrap, P. M., Adams, I. P., Peters, J. C., Budge, G. E., & Boonham, N. (2012). A new large scale soil DNA extraction procedure and real-time PCR assay for the detection of Sclerotium cepivorum in soil. European Journal of Plant Pathology, 134, 467–473.CrossRefGoogle Scholar

Copyright information

© UK Crown Copyright 2013

Authors and Affiliations

  • James W. Woodhall
    • 1
  • Ian P. Adams
    • 1
  • Jeff C. Peters
    • 1
  • Glyn Harper
    • 2
  • Neil Boonham
    • 1
  1. 1.The Food and Environment Research AgencyYorkUK
  2. 2.AHDB Potato Council, Sutton Bridge Crop Storage ResearchSpaldingUK

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