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@ARTICLE{TreeBASE2Ref19017, author = {Frank N Martin}, title = {Mitochondrial haplotype analysis as a tool for differentiating isolates of Verticillium dahliae.}, year = {2010}, keywords = {}, doi = {}, url = {http://}, pmid = {}, journal = {Phytopathology}, volume = {100}, number = {}, pages = {}, abstract = {The ability to monitor mitochondrial background in Verticillium dahliae may provide an additional tool for population studies and monitoring clonal populations. Published mitochondrial genome sequences of V. dahliae (DQ351941) were used to design primers for amplification of spacer regions for assessment of mitochondrial haplotype differences among isolates. Five regions were examined (5,229 bp, or 19% of the total genome size) for 30 isolates representing a range in VCG, host, and geographic origin. Observed differences among isolates were due to single nucleotide polymorphisms, different numbers of bases in specific homopolymeric regions, and copies of subrepeated sequences. When all the differences observed for each locus was totaled there were polymorphisms at 28 loci; when the results of each locus for individual isolates were combined there were 15 mitochondrial haplotypes. Some of the observed grouping correlated with VCG. For example, five VCG-1A and ?1B isolates from California, Spain and Greece had identical haplotypes, however, this was not observed for VCG-2 or VCG-4 isolates. While some VCG-2 isolates also were identical and fell into a single haplotype, 5 haplotypes were found for this group (5 other haplotypes were observed for other isolate that had not been characterized to VCG but grouped with VCG-2 isolates in the phylogenetic analysis). Likewise, five VCG-4 isolates fell into 4 mitochondrial haplotypes, one of which was identical to the largest VCG-2 grouping. A heterokaryon self-incompatible isolate that was reported in the literature to cluster with VCG-2 isolates by AFLP analysis grouped was identical with VCG-1 isolates for 4 of the 5 loci but was intermediate between VCG-1 and VCG-2 in the haplotype analysis. Phylogenetic analysis with these five regions revealed the mitochondrial background of VCG-1 and VCG2-B to be monophyletic but VCG-2A and VCG-4 could not be separated. The results obtained indicate that there is variation in mitochondrial haplotypes and this type of analysis may be a useful for characterization of isolates. While data from all five regions was needed for the haplotype separation obtained in this study, depending on the VCG or the level of variability observed within a population it may be possible to use fewer loci.} }