, Volume 163, Issue 2, pp 105–115 | Cite as

Intraspecific variation of Rhizoctonia solani AG 3 isolates recovered from potato fields in Central Iran and South Australia

  • G. R. Balali
  • S. M. Neate
  • A. M. Kasalkheh
  • B. J. Stodart
  • D. L. Melanson
  • E. S. Scott


Pectic zymogram, RFLP and PCR analyses were used to characterize Rhizoctonia solani AG 3 isolates collected from diseased potatoes in South Australia. The pectic zymogram data were compared with those obtained for isolates collected from central Iran. Analyses of bands corresponding to pectin esterase and polygalacturonase revealed three zymogram subgroups (ZG) in AG 3. In addition to the previously reported ZG7 (here renamed ZG7-1), two new zymogram subgroups, ZG7-2 and ZG7-3, were identified. Of the 446 isolates tested, 50% of the South Australian and 46% of the Iranian isolates were ZG7-1. The majority of the isolates originating from stem and root cankers were ZG7-1, whereas most of the isolates designated ZG7-2 and ZG7-3 originated from tuber-borne sclerotia. Pathogenicity tests revealed that ZG7-1 generally produced fewer sclerotia and more severe cankers of underground parts of the potato plants than the other two ZGs. Two random DNA clones, one originating from an AG 3 isolate and the other from an AG 4 isolate, were used as probes for RFLP analyses of Australian isolates. The AG 3 probe, previously identified to be specific to this group, detected a high level of genetic diversity, with 11 genotypes identified amongst 50 isolates analysed. The low-copy AG 4 probe resolved three genotypes amongst 24 isolates. For 23 isolates analysed with both markers, the combined data distinguished a total of six genotypes and similarity analysis resolved the isolates into two main groups with 50% homology. PCR, using primers for the plant intron splice junction region (R1), also revealed variation. No obvious relationship among pectic zymogram groups, RFLP and PCR genotypes was observed.


black scurf genetic variation pectic isozymes potato Rhizoctonia 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Adams GC (1988). Adv Plant Pathol 6: 535–552 Google Scholar
  2. 2.
    Micales JA, Bonde MR and Peterson GL (1986). The use of isozyme analysis in fungal taxonomy and genetics. Mycotaxon 27: 405–449 Google Scholar
  3. 3.
    Liu ZL and Sinclair JB (1992). Genetic diversity of Rhizoctonia solani anastomosis group 2. Phytopathology 82: 778–787 CrossRefGoogle Scholar
  4. 4.
    Schneider JHM, Salazar O, Rubio V and Keijer J (1997). Identification of Rhizoctonia solani associated with field-grown tulips using ITS rDNA polymorphism and pectic enzymes. Eur J Plant Pathol 103: 607–622 CrossRefGoogle Scholar
  5. 5.
    Clark J, Butters J, Brent KJ and Hollomon DW (1989). Isozyme uniformity in Erysiphe graminis f.sp. hordei. Mycol Res 92: 404–409 CrossRefGoogle Scholar
  6. 6.
    Goodwin SB, Webster RK and Allard RW (1994). Phytopathology 84: 1047–1053 CrossRefGoogle Scholar
  7. 7.
    Meijer G, Megnegneau B and Linders EGA (1994). Mycol Res 98: 267–276 CrossRefGoogle Scholar
  8. 8.
    McDonald BA (1997). The population genetics of fungi: Tools and techniques. Phytopathology 87: 448–453 CrossRefPubMedGoogle Scholar
  9. 9.
    Stummer BE, Zanker T, Scott ES and Whisson DL (2000). Genetic diversity in populations of Uncinula necator: Comparison of RFLP and PCR-based approaches. Mycol Res 104: 44–52 CrossRefGoogle Scholar
  10. 10.
    Duncan S, Barton JE and O’Brien PA (1993). Analysis of variation in isolates of Rhizoctonia solani by random amplified polymorphic DNA assay. Mycol Res 97: 1075–1082 CrossRefGoogle Scholar
  11. 11.
    Salazar O, Schneider JHM, Julián MC, Keijer J and Rubio V (1999). Phylogenetic subgrouping of Rhizoctonia solani AG 2 isolates based on ribosomal ITS sequences. Mycologia 91: 459–467 CrossRefGoogle Scholar
  12. 12.
    Cruickshank RH (1990). Mycol Res 94: 938–946 CrossRefGoogle Scholar
  13. 13.
    MacNish GC and Sweetingham MW (1993). Evidence of stability of pectic zymogram groups within Rhizoctonia solani AG-8. Mycol Res 97: 1056–1058 CrossRefGoogle Scholar
  14. 14.
    Neate SM, Cruickshank RH and Rovira AD (1988). Pectic enzyme patterns of Rhizoctonia solani isolates from agricultural soils in South Australia. Trans Br Mycol Soc 90: 37–42 CrossRefGoogle Scholar
  15. 15.
    Sweetingham MW, Cruickshank RH and Wong DH (1986). Trans Br Mycol Soc 86: 305–311 CrossRefGoogle Scholar
  16. 16.
    MacNish GC, Carling DE, Sweetingham MW and Brainard KA (1994). Anastomosis group (AG) affinity of pectic isozyme (zymogram) groups (ZG) of Rhizoctonia solani from the Western Australia cereal-belt. Mycol Res 98: 1369–1375 CrossRefGoogle Scholar
  17. 17.
    Jabaji-Hare SH, Meller Y, Gill S and Charest PM (1990). Investigation of genetic relatedness among anastomosis groups of Rhizoctonia solani using cloned DNA probes. Can J Plant Pathol 12: 393–404 CrossRefGoogle Scholar
  18. 18.
    Balali GR, Whisson DL, Scott ES and Neate SM (1996). DNA fingerprinting probe specific to isolates of Rhizoctonia solani AG-3. Mycol Res 100: 467–470 CrossRefGoogle Scholar
  19. 19.
    Matthew J and Whisson D (1995). DNA probe specific to Rhizoctonia solani anastomosis group 8. Mycol Res 99: 745–750 CrossRefGoogle Scholar
  20. 20.
    Weining S and Langridge P (1991). Identification and mapping of polymorphisms in cereals based on the polymerase chain reaction. Theor Appl Genet 182: 209–216 Google Scholar
  21. 21.
    Kuninaga S, Natsuaki T, Takeuchi T and Yokosawa R (1997). Curr Genet 32: 237–243 CrossRefPubMedGoogle Scholar
  22. 22.
    Pope EJ and Carter DA (2001). Phylogenetic placement and host specificity of mycorrhizal isolates belonging to AG-6 and AG-12 in the Rhizoctonia solani species complex. Mycologia 93: 712–719 CrossRefGoogle Scholar
  23. 23.
    Guillemaut C, Edel-Hermann V, Camporota P, Alabouvette C, Richard-Molard M and Steinberg C (2003). Typing of anastomosis groups of Rhizoctonia solani by restriction analysis of ribosomal DNA. Can J Microbiol 49: 556–568 CrossRefPubMedGoogle Scholar
  24. 24.
    Balali GR, Neate SM, Scott ES, Whisson DL and Wicks TJ (1995). Anastomosis group and pathogenicity of isolates of Rhizoctonia solani from potato crops in South Australia. Plant Pathol 44: 1050–1057 CrossRefGoogle Scholar
  25. 25.
    Parmeter JR, Sherwood RT and Platt WD (1969). Phytopathology 59: 1270–1278 Google Scholar
  26. 26.
    Cruickshank RH and Wade GC (1980). Detection of pectic enzymes in pectin-acrylamide gels. Anal Biochem 107: 177–181 CrossRefPubMedGoogle Scholar
  27. 27.
    Cruickshank RH (1983). Distinction between Sclerotinia species by their pectic zymograms. Trans Br Mycol Soc 80: 117–119 CrossRefGoogle Scholar
  28. 28.
    Balali GR. Genetic variation of Rhizoctonia solani AG-3 in South Australia. PhD thesis. The University of Adelaide, Australia, 1996; 189 ppGoogle Scholar
  29. 29.
    Sambrook J, Fritsch EF and Maniatis T (1989). Molecular Cloning: A Laboratory Manual. (2nd ed.). Cold Spring Harbor, New York Google Scholar
  30. 30.
    Hollander M and Wolfe DA (1999). Nonparametric Statistical Methods. John Wiley and Sons, NY, 800 Google Scholar
  31. 31.
    Lynch M (1990). The similarity index and DNA fingerprinting. Mol Biol Evol 7: 478–484 PubMedGoogle Scholar
  32. 32.
    Nei M and Li WH (1979). Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Natl Acad Sci U S A 76: 5269–5273 CrossRefPubMedGoogle Scholar
  33. 33.
    Laroche JP, Jabaji-Hare SH and Charest PM (1992). Differentiation of two anastomosis groups of Rhizoctonia solani by isozyme analysis. Phytopathology 82: 1387–1393 CrossRefGoogle Scholar
  34. 34.
    Yang HA, Zhou J, Sivasithamparam K, Tommerup IC, Barton JE and O’Brien PA (1994). Genetic variability in pectic enzymes of Rhizoctonia solani isolates causing bare-patch disease of cereals. J Phytopathol 141: 259–266 CrossRefGoogle Scholar
  35. 35.
    O’Brien PA, Vidyani M, Zamani MR. Pectic enzymes as virulence factors in Rhizoctonia solani AG 8 strains. International Symposium on Rhizoctonia (Proceeding) Noordwijkerhout, the Netherlands June 1995: 3–4Google Scholar
  36. 36.
    Barker KR and Walker JC (1962). Relationship of pectolytic and cellulytic enzyme production by strains of Pellicularia filamentosa to their pathogenicity. Phytopathology 52: 1119–1125 Google Scholar
  37. 37.
    Gudmestad NC, Zink RT and Huguelet JE (1979). The effect of harvest date and tuber-borne sclerotia on the severity of Rhizoctonia disease of potato. Am Potato J 56: 35–41 CrossRefGoogle Scholar
  38. 38.
    James WC and McKenzie AR (1972). The effect of tuber-borne sclerotia of Rhizoctonia solani Kuhn on the potato crop. Am Potato J 49: 296–301 CrossRefGoogle Scholar
  39. 39.
    Weinhold AR, Bowman T and Hall DH (1982). Rhizoctonia disease of potato: Effect on yield and control by seed tuber treatment. Plant Dis 66: 815–818 CrossRefGoogle Scholar
  40. 40.
    Frank JA (1981). Hooker, WJ (eds) Compendium of Potato Diseases, pp 52–54. APS Press, Minnesota, USA Google Scholar

Copyright information

© Springer 2007

Authors and Affiliations

  • G. R. Balali
    • 1
    • 2
  • S. M. Neate
    • 3
  • A. M. Kasalkheh
    • 1
  • B. J. Stodart
    • 2
  • D. L. Melanson
    • 4
  • E. S. Scott
    • 2
  1. 1.Department of BiologyThe University of IsfahanIsfahanIran
  2. 2.School of Agriculture, Food and WineThe University of AdelaideGlen OsmondAustralia
  3. 3.Department of Plant PathologyNorth Dakota State UniversityFargoUSA
  4. 4.Formerly South Australian Research and Development InstituteAdelaideAustralia

Personalised recommendations