Typing of anastomosis groups of Rhizoctonia solani by restriction analysis of ribosomal DNA

• Published in: Canadian journal of microbiology Vol. 49; no. 9; pp. 556 - 568
• Main Authors: Guillemaut, C, Edel-Hermann, V, Camporota, P, Alabouvette, C, Richard-Molard, M, Steinberg, C
• Format: Journal Article
• Language: English
• Published: Ottawa, Canada NRC Research Press 01.09.2003; National Research Council of Canada; Canadian Science Publishing NRC Research Press
• Subjects: Biological and medical sciences; Crop diseases; Disease; DNA, Fungal - analysis; DNA, Fungal - genetics; DNA, Ribosomal - analysis; DNA, Ribosomal - genetics; Enzymes; Flowers & plants; Fundamental and applied biological sciences. Psychology; Fungi; Genetics; internal transcribed spacers; Life Sciences; Microbiology; Microbiology and Parasitology; Miscellaneous; molecular sequence data; Mycological Typing Techniques; Mycology; nucleotide sequences; Plant diseases; plant pathogenic fungi; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; restriction fragment length polymorphism; Rhizoctonia - classification; Rhizoctonia - genetics; Rhizoctonia solani; Ribonucleic acid; ribosomal DNA; Ribotyping; RNA; Roots (Botany); strain differences; Sugar; Sugar beets; Thanatephorus cucumeris; France; Fungi; Restriction fragment length polymorphism; Typing; Plant pathogen; Ribosomal DNA; Rhizoctonia solani; Identification; Fungi Imperfecti; Thallophyta
• ISSN: 0008-4166; 1480-3275
• DOI: 10.1139/w03-066

A method based on restriction analysis of polymerase chain reaction (PCR)-amplified ribosomal DNA was developed for the rapid characterization of large populations of Rhizoctonia solani at the anastomosis group (AG) level. The restriction maps of the internal transcribed spacers (ITS) sequences were compared for 219 isolates of R. solani belonging to AG-1 to AG-12 and AG-BI, representing diverse geographic and host range origins. Four discriminant restriction enzymes (MseI, AvaII, HincII, and MunI) resolved 40 restriction fragment length polymorphism (RFLP) types among the 219 ITS sequences of R. solani. Each RFLP type could be assigned to a single AG except for two RFLP types, which were common to two AG. A fifth enzyme allowed the discrimination of AG-6 and AG-12. In addition, the combination of four enzymes allowed the discrimination of subsets within AG-1, AG-2, AG-3, and AG-4. The efficiency of the typing method was confirmed by analyzing PCR-amplified ITS sequences of 30 reference strains. Furthermore, the PCR–RFLP method was used to characterize at the AG level 307 isolates of R. solani originating from ten sugar beet fields exhibiting patches of diseased plants in France. The PCR-based procedure described in this paper provides a rapid method for AG typing in R. solani.Key words: Rhizoctonia solani, anastomosis group, PCR–RFLP, ITS, identification, sugar beet.