Molecular characterization of the genome of a partitivirus from the basidiomycete Rhizoctonia solani

The bisegmented genome of a double-stranded (ds) RNA virus from the fungus Rhizoctonia solani isolate Rhs 717 was characterized. The larger segment, dsRNA 1, is 2363 bases long whereas the smaller segment, dsRNA 2, has 2206 bases. The 5′ ends of the coding strands of dsRNA 1 and dsRNA 2 are highly conserved (100% identity over 47 bases), and contain inverted repeats capable of forming stable stem–loop structures. Analysis of the coding potential of each of the two segments showed that dsRNAs 1 and 2 could code for polypeptides of 730 aa (bases 86–2275; molecular mass 86 kDa) and 683 aa (bases 79–2130; molecular mass 76 kDa), respectively. The 86 kDa polypeptide has all the motifs of dsRNA RNA-dependent RNA polymerases (RDRP), and has significant homology with putative RDRPs of partitiviruses from Fusarium poae and Atkinsonella hypoxylon. The 76 kDa protein shows homology with the putative capsid proteins (CP) of the same viruses. Northern blot analysis revealed no subgenomic RNA species, consistent with the fact that the long open reading frames encoding the putative RDRP and CP cover the entire length of the respective dsRNAs.

Isozyme variation in the fungus Atkinsonella hypoxylon within and among populations of its host grasses

Isozyme variation of 291 isolates of Atkinsonella hypoxylon (Clavicipitaceae, tribe Balansieae) from 24 populations of its four known host grasses (Danthonia compressa, Danthonia sericea, Danthonia spicata, and Stipa leucotricha) was examined using starch gel electrophoresis. In total, there were 20 distinct multilocus genotypes. Eleven out of 13 enzyme loci (84.6%) exhibited more than one allele (mean 2.8) per locus. Nei's total gene diversity (HT) within all isolates was 0.229. Between isolate samples from S. leucotricha and the three Danthonia hosts, Nei's genetic identity (I) ranged from 0.21 to 0.31 and among isolate samples from the three Danthonia species I ranged from 0.65 to 0.88, with isolates from D. spicata and D. compressa being most similar. Variation of A. hypoxylon occurred both within and among populations of D. spicata and D. compressa, where up to 53 isolates were sampled per host population. In contrast, all 20 isolates from S. leucotricha were identical, as were all 6 from D. sericea. A few isolates from D. spicata exhibited the same, unusual multilocus genotype with unique alleles at six different loci. The occurrence of several multilocus genotypes in isolates from the same ascostroma and the 1:1 segregation of genotypes among ascospores from a single ascus indicated gene exchange among sexually reproducing individuals, consistent with a heterothallic mating system for A. hypoxylon.