Hyphal anastomosis reactions, rDNA-internal transcribed spacer (ITS) sequences, and virulence of isolates representing Rhizoctonia solani AG-BI and six subsets of anastomosis group (AG)-2 (-2-1, -2-2 IIIB, -2-2 IV, -2-2 LP, -2-3, and -2-4) were compared. AG-2-4 is a subset described for the first time in this report. Anastomosis reactions within AG-BI and the listed subsets of AG-2 were generally strong but, between subsets, ranged from strong to a very weak "bridging" -type reaction. Anastomosis reaction alone generally did not provide adequate evidence for placement of an isolate into a subset of AG-2. Anastomosis reactions between AG-BI and the original subsets of AG-2 (-2-1 and -2-2) are very strong; for this reason, we propose that it be included as a subset of AG-2 (designation AG-2 BI). Subsets -2-3 and -2-4 show very weak bridging-type anastomosis reactions with all other subsets of AG-2 and thus may be candidates for independent AG status. Grouping within AG-2 based on rDNA-ITS sequences was consistent with the abovementioned subsets. However, grouping based on virulence as measured herein does not conform to established grouping patterns within AG-2 and does not seem useful as a group-defining criterion. A broad range of damage was observed among members of the most virulent subsets (-2-1, -2-2 IIIB, -2-2 IV, and -2-4), whereas other subsets (-2 BI, -2-2 LP, and -2-3) were similar to one another in causing a minimal level of damage. Group-specific primer pairs for each of the seven subsets of AG-2 were designed based on the abovementioned rDNA-ITS sequences. Primer pairs proved dependable and subset specific in polymerase chain reaction amplifications of purified genomic DNA from 109 isolates of R. solani and two isolates of binucleate Rhizoctonia. These primers will provide a simple and useful method for subset-specific characterization within AG-2 if further critical evaluations confirm their specificity.

Sequence analysis of the rDNA region containing the internal transcribed spacer (ITS) regions and the 5.8s rDNA coding sequence was used to evaluate the genetic diversity of 45 isolates within and between anastomosis groups (AGs) in Rhizoctonia solani. The 5.8s rDNA sequence was completely conserved across all the AGs examined, whereas the ITS rDNA sequence was found to be highly variable among isolates. The sequence homology in the ITS regions was above 96% for isolates of the same subgroup, 66-100% for isolates of different subgroups within an AG, and 55-96% for isolates of different AGs. In neighbor-joining trees based on distances derived from ITS-5.8s rDNA sequences, subgroups IA, IB and IC within AG-1 and subgroups HG-I and HG-II within AG-4 were placed on statistically significant branches as assessed by bootstrap analysis. These results suggest that sequence analysis of ITS rDNA regions of R. solani may be a valuable tool for identifying AG subgroups of biological significance.

Classifi cation of Rhizoctonia spp. using rDNA-ITS sequence analysis supports the genetic basis of the classical anastomosis grouping Abstract Currently, rDNA-ITS sequence analysis seems to be the most appropriate method for comprehensive classifi cation of Rhizoctonia spp. Our previous review article was concerned with detailed analysis of multinucleate Rhizoctonia (MNR), and the current review complements the previous one with detailed analysis of binucleate Rhizoctonia (BNR) (teleomorphs: Ceratobasidium spp. and Tulasnella spp.) and uninucleate Rhizoctonia (UNR) (teleomorph: C. bicorne). Data of all the appropriate BNR and UNR accumulated in GenBank were analyzed together in neighborjoining (NJ) trees supplemented with percent sequence similarity within and among the anastomosis groups (AGs) and subgroups. Generally, the clusters of the isolate sequences supported the genetic basis for the AG based on hyphal fusion anastomosis. Comprehensive interrelationships among all the currently available MNR, BNR, and UNR groups and subgroups in GenBank were subsequently analyzed in NJ and maximum-parsimony (MP) trees, showing the genetic relatedness among the different groups and indicating possible bridging groups between MNR, BNR, and UNR. The review also indicates serious inaccuracies in designation of sequences of some isolates deposited in GenBank. Several additional teleomorph genera with Rhizoctonia spp. anamorphs have also been reported in the literature. However, as they have not been intensively studied, there were no available data on their rDNA-ITS sequences that could be included in this review.

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