Hyphal Anastomosis Reactions, rDNA-Internal Transcribed Spacer Sequences, and Virulence Levels Among Subsets of Rhizoctonia solani Anastomosis Group-2 (AG-2) and AG-BI

ABSTRACT 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.