Typing of anastomosis groups ofRhizoctonia solaniby restriction analysis of ribosomal DNA

2003 ◽  
Vol 49 (9) ◽  
pp. 556-568 ◽  
Author(s):  
Cécile Guillemaut ◽  
Véronique Edel-Hermann ◽  
Pierre Camporota ◽  
Claude Alabouvette ◽  
Marc Richard-Molard ◽  
...  
Keyword(s):  
Sugar Beet ◽  
Rhizoctonia Solani ◽  
Ribosomal Dna ◽  
Restriction Analysis ◽  
Anastomosis Group ◽  
Internal Transcribed Spacers ◽  
Its Sequences ◽  
Typing Method ◽  
Rflp Type ◽  
Pcr Rflp

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.

scholarly journals Identification of Rhizoctonia solani isolates from sugar beet roots by analyzing the ITS region of ribosomal DNA

2007 ◽  
pp. 161-171 ◽  
Author(s):  
Vera Stojsin ◽  
Dragana Budakov ◽  
Barry Jacobsen ◽  
Eva Grimme ◽  
Ferenc Bagi ◽  
...  
Keyword(s):  
Sugar Beet ◽  
Rhizoctonia Solani ◽  
Ribosomal Dna ◽  
Root Rot ◽  
Its Region ◽  
Anastomosis Group ◽  
Crown Rot ◽  
Internal Transcribed Spacer Region ◽  
The Usa ◽  
Crown And Root Rot

Rhizoctonia solani (K?hn) is one of the most important sugar beet pathogens Rhizoctonia solani anastomosis groups (AGs) 2-2 and 4 are proven to be the most common pathogenic strains on sugar beet. AG 2-2 (intraspecific groups IIIB and IV) can cause root and crown rot while damping-off of seedlings is most frequently attributed to AG 4. Four isolates of R. solani from sugar beet roots showing characteristic crown and root rot symptoms, collected from different localities in Vojvodina Province, were chosen and compared to the well-characterized R. solani isolate R9, AG 2-2 IV, from the USA. All Vojvodinian isolates showed medium level of pathogenicity and were able to cause crown and root rot symptoms on inoculated sugar beet roots. Based on anastomosis reaction, isolates from Vojvodina did not belong to the AG 2-2 group. Sequencing of the ITS (internal transcribed spacer) region of ribosomal DNA was performed on the Vojvodinian isolates from R9 in order to determine their relatedness. Sequence analysis showed that these isolates were different than R9 and were closely related (99-100% sequence homology) to anastomosis group 4, subgroup HG II.

Rapid Molecular Typing of Methicillin-ResistantStaphylococcus aureusby PCR-RFLP

2001 ◽  
Vol 22 (5) ◽  
pp. 294-298 ◽  
Author(s):  
Thomas A. Wichelhaus ◽  
Klaus-Peter Hunfeld ◽  
Boris Böddinghaus ◽  
Peter Kraiczy ◽  
Volker Schàfer ◽  
...  
Keyword(s):  
Fragment Length Polymorphism ◽  
Restriction Analysis ◽  
Protein A ◽  
Hypervariable Region ◽  
Epidemiological Surveillance ◽  
Typing Method ◽  
Rapid Pcr ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Rflp Typing

AbstractObjective:To establish a new, rapid, and reliable genotypic fingerprinting technique for methicillin-resistantStaphylococcus aureus(MRSA) typing in routine epidemiological surveillance.Design:The method is based on polymerase chain reaction (PCR) restriction fragment-length polymorphism (RFLP) followingHaeII digestion of simultaneously amplified parts of the protein A gene, the coagulase gene, and the hypervariable region adjacent tomecA. A total of 46 MRSA initial isolates were analyzed, including 14 isolates from five countries; the six German epidemic strains; 16 isolates from the Frankfurt metropolitan area, which were known to be heterogeneous by pulsed-field gel electrophoresis (PFGE); and 10 isolates obtained during three epidemics, all of which displayed an identical genotype.Results:Restriction analysis by PCR-RFLP permitted discrimination of 10 of 14 international isolates, all six German epidemic strains, and 15 of 16 national isolates. It also confirmed the homogeneous character of the 10 outbreak isolates.Conclusions:This new and rapid PCR-RFLP typing method is an attractive tool in routine epidemiological surveillance. Its impressive characteristics are ease of performance and interpretation, while at the same time guaranteeing good discriminatory power, reproducibility, and typeability.

Analysis of intragenomic variation of the rDNA internal transcribed spacers (ITS) in Halichondrida (Porifera: Demospongiae)

2007 ◽  
Vol 87 (6) ◽  
pp. 1599-1605 ◽  
Author(s):  
Belinda Alvarez ◽  
Mahima Krishnan ◽  
Karen Gibb
Keyword(s):  
Genetic Markers ◽  
Ribosomal Dna ◽  
Internal Transcribed Spacers ◽  
Its Sequences ◽  
Single Individual ◽  
Closely Related Species ◽  
Hybrid Species ◽  
Mitochondrial Markers ◽  
Intragenomic Variation ◽  
Acanthella Cavernosa

Three species of sponges, Axinella aruensis, Phakellia sp. and Acanthella cavernosa, were selected to investigate the extent of intragenomic variation (IGV) in the internal transcribed spacers (ITS-1 and ITS-2) of the ribosomal DNA within the order Halichondrida. The IGV was detected in both the spacers and in the three species. At least three non-identical sequences were found in any single individual with the number of polymorphic sites within species ranging from 6 to 77. Uncorrected distance values up to 29% were observed in the ITS-2 of Axinella aruensis. The levels of polymorphism found in this species are the highest reported for Porifera and comparable to those observed in hybrid species of scleractinean corals. Analyses of ITS sequences from multiple clones from individuals of Halichondrida are strongly recommended if these genetic markers are to be used to depict phylogenetic and/or phylogeographic relationships of closely related species and their populations. Data from other nuclear and mitochondrial markers should be used to complement conclusions derived from studies based on ITS sequences.

scholarly journals Differentiation of Three Homogeneous Groups of Rhizoctonia solani Anastomosis Group 4 by Analysis of Fatty Acids

2001 ◽  
Vol 91 (9) ◽  
pp. 821-830 ◽  
Author(s):  
Janell Stevens Johnk ◽  
Roger K. Jones
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Sugar Beet ◽  
North Dakota ◽  
Rhizoctonia Solani ◽  
Acid Methyl Ester ◽  
Anastomosis Group ◽  
Homogeneous Groups ◽  
Fame Analysis ◽  
Group 4

Profiles of fatty acids from 70 isolates of Rhizoctonia solani anastomosis group (AG)-4 clustered into three groups, corresponding to homogeneous group (HG)-I, HG-II, and a newly described HG-III. Isolates from Georgia peanuts exhibiting limb rot were characterized as gas chromatography (GC) subgroup 1 (GC-1) and contained HG-I isolates. Isolates from diseased soybean hypocotyls grown in North Dakota and sugar beet seedlings, taproots, and tare soil in Minnesota and North Dakota were characterized as GC subgroup 2 (GC-2) and contained predominantly HG-II isolates but also included three distinct isolates based on fatty acid methyl ester (FAME) analysis and morphological features. Selected isolates from North Carolina cucumbers clustered into three distinct groups that corresponded to HG-I, HG-II, and the newly described HG-III. Distinct isolates from the soybean and sugar beet populations clustered with HG-III. Fatty acid profiles of AG-4 were compared with FAME library profiles of AG-1, AG-2 type 2, and AG-3, which were developed in previous studies and were sufficiently different that they could be used to support speciation of this group from R. solani. It is suggested that binomial R. practicola may be appropriate for the portion of AG-4 identified as HG-II.

scholarly journals Influence of selected Rhizoctonia solani isolates on sugar beet seedlings

2016 ◽  
Vol 56 (2) ◽  
pp. 116-121
Author(s):  
Paweł Skonieczek ◽  
Mirosław Nowakowski ◽  
Jacek Piszczek ◽  
Marcin Żurek ◽  
Łukasz Matyka
Keyword(s):  
Sugar Beet ◽  
Rhizoctonia Solani ◽  
Root Rot ◽  
Laboratory Tests ◽  
Anastomosis Group ◽  
Damping Off ◽  
Highly Pathogenic ◽  
Breeding Lines ◽  
Brown Root Rot

Abstract From 2008 to 2010 the levels of sugar beet seedlings infection caused by Rhizoctonia solani were compared in laboratory tests. Seven sugar beet lines were tested: H56, H66, S2, S3, S4, S5 and S6 as well as three control cultivars: Carlos, Esperanza and Janosik. Sugar beet lines with tolerance to rhizoctoniosis and cultivars without tolerance were infected artificially by R. solani isolates: R1, R28a and R28b. These isolates belong to the second anastomosis group (AG), which is usually highly pathogenic to beet roots. The aim of the experiment was to test whether the tolerance of sugar beet genotypes to R. solani AG 2 prevents both root rot, and damping-off of seedlings, induced by the pathogen. Sugar beet lines tolerant to brown root rot in laboratory tests were significantly less sensitive to infection of the seedlings by R. solani AG 2 isolates in comparison to control cultivars. Rhizoctonia solani AG 2 isolates demonstrated considerable differences in pathogenicity against seedlings of sugar beet lines and cultivars. The strongest infection of sugar beet seedlings occurred with the isolate R28b. The greatest tolerance to infection by AG 2 isolates was found for the S5 and S3 breeding lines.

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