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1- .2-.3- .*-:[email protected]

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References1. Anderson NA. 1982. The genetics and pathology of Rhizoctonia solani. Annual

Review of Phytopathology 20: 329–347.2. Balali G and Kowsari M, 2004. Pectic zymogram variation and pathogenicity of

Rhizoctonia solani AG-4 to bean (Phaseolus vulgaris) isolates in Isfahan, Iran.Mycopathologia 158: 377–384.

3. Boysen M, Borja M, Del Moral C, Salazar O and Rubio V. 1996. Identification atstrain level of Rhizoctonia solani AG4 isolates by direct sequence of asymmetricPCR products of the ITS regions. Current Genetics 29: 174–181.

4. Bruns TD, White TJ and Taylor JW. 1991. Fungal molecular systematics. AnnualReview of Ecology, Evolution and Systematics 22: 525–564.

5. Cubeta MA, Vilgalys R and González D. 1996. Molecular analysis of ribosomalRNA genes in Rhizoctonia fungi. pp. 81–86, In B Sneh, S Jabaji-Hare, S Neate andG Dijst (eds). Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology,Pathology and Disease Control. Dordrecht: Kluwer Academic Publishers.

6. González D, Carling DE, Kuninaga S, Vilgalys R and Cubeta MA. 2001.Ribosomal DNA systematics of Ceratobasidium and Thanatephorus withRhizoctonia anamorphs. Mycologia 93: 1138–1150.

7. González D, Cubeta MA and Vilgalys R. 2006. Phylogenetic utility of indels withinribosomal DNA and -tubulin sequences from fungi in the Rhizoctonia solanispecies complex. Molecular Phylogenetics and Evolution 40: 459–470.

8. González V, Salazar O, Julián MC, Acero J, Portal MA, Muñoz R, López-CórcolesH, Gómez-Acebo E, López-Fuster P and Rubio V. 2002. Ceratobasidiumalbasitensis, a new Rhizoctonia-like fungus isolated in Spain. Persoonia 17: 601–614.

9. Guillemaut C, Edel-Herman V, Comporota P, Alabouvette C, Richard-Molard Mand Steinberg C. 2003. Typing of anastomosis groups of Rhizoctonia solani byrestriction analysis of ribosomal DNA. Canadian Journal of Microbiology 49: 556–568.

10. Haratian M. 2012. Evaluation genetic structure populations Rhizoctonia solani AG4in Iran [PhD]. [Tehran]: Tarbiat Modares University.

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11. Jabaji-Hare S. 1996. Biochemical methods. pp. 65–71, In B Sneh, S Jabaji-Hare, SNeate and G Dijst (eds). Rhizoctonia Species: Taxonomy, Molecular Biology,Ecology, Pathology and Disease Control. Dordrecht: Kluwer Academic Publishers.

12. Kiliço lu Mc and Özkoç I. 2010. Molecular characterization of Rhizoctonia solaniAG4 using PCR-RFLP of the rDNA-ITS region. Turkish. Journal of Biology 34:261–269.

13. Kuninaga S 1996. DNA base sequence relatedness. pp. 74–78, In RhizoctoniaSpecies: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control.(B Sneh, S Jabaji-Hare, S Neate and G Dijst (eds). Dordrecht: Kluwer AcademicPublishers.

14. Kuninaga S and Yokosawa R. 1984a. DNA base sequence homology inRhizoctonia solani Ktihn. IV. Genetic relatedness within AG4. Annals of thePhytopathological society of Japan 50: 322–330.

15. Kuninaga S and Yokosawa R. 1984b. DNA base sequence homology inRhizoctonia solani Ktihn. V. Genetic relatedness within AG6. Annals of thePhytopathological society of Japan 50: 346–352.

16. Lübeck M. 2004. Molecular characterization of Rhizoctonia solani. AppliedMycology and Biotechnology 4: 205–224.

17. McDonald HJ and Rovira AD. 1985. Development of inoculation technique forRhizoctonia solani and its application to screening cereal cultivars for resistance.pp. 174–176, In CA Parker, AD Rovira, KJ Moore, PTW Wong and JF Kollmorgen(eds.). Ecology and Management of Soilborne Plant Pathogens. St. Paul (MN):American Phytopathological Society.

18. Meinhardt LW, Wulff NA, Bellato CM and Tsai SM. 2002. Genetic analyses ofRhizoctonia solani isolates from Phaseolus vulgaris grown in the Atlanticrainforest region of Sao Paulo, Brazil. Fitopatologia Brasileira 27: 259–267.

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Research in Plant Pathology/Vol. 5/No. 1/Spring and Summer 2017 2

Study on DNA polymorphism in populations of Rhizoctonia solanianastomosis group 4 using rDNA RFLP

F. Badpa*1, G.R. Balali 2, B. Sharifnabi3

AbstractRibosomal DNA (rDNA) sequences have been widely used to study the phylogenetic

relationships in different fungi. Fungal nuclear rRNA genes are arranged as tandem repeatswith several hundred copies per genome. These spacer regions are considerably more variablethan the subunit sequences and have been widely used in studies on the relationships amongspecies within a single genus or among intraspecific populations. To evaluate thepolymorphism between 18S and 28S genes, 28 isolates of Rhizoctonia solani anastomosisgroup 4 were examined. Genomic DNA was extracted from the isolates and prepared for PCRreaction. The amplification was performed using internal transcribed spacer (ITS) ITS1 andITS4 primers. A DNA fragment of 700 bp in size was detected in PCR products of all testedisolates. To assess existence of any further polymorphism in the ITS region, the PCR productswere digested with restriction endonucleases. Although there were restriction sites for XbaI,HaeIII, BamHI, HindIII, SacI, PstI, and TaqI endonucleases, there were no restriction sites forNdeI, XhoI, HincII, HinfI, EcoRI and XhoI endonucleases. The endonuclease HincIIrecognized a restriction site on PCR products that discriminated the isolates belonging toAG4-HGII form isolates of AG4-HGI. Based on the results, it has been concluded that AG-4isolates of Rhizoctonia solani were heterogenic.

Keywords: Genetic diversity, ITS-rDNA, PCR, Rhizoctonia solani AG4.

1 - Former MSc student, Department of Biology, Isfahan University, Isfahan, Iran.2- Associate professor, Department of Biology, Isfahan University, Isfahan, Iran.3- Professor, Department of Plant Protection, College of Agriculture, Isfahan University of Technology,

Isfahan, Iran.*Corresponding author: [email protected]

mailto:[email protected]

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