No abstract

It has been suggested that Candida albicans, a diploid asexual fungus, achieves genetic diversity by genomic rearrangement. This important human pathogen may provide a system in which to analyze alternate routes to genomic diversity. C. albicans has a highly variable karyotype; its chromosomes contain a middle repeated DNA sequence called the Major Repeat Sequence (MRS), composed of subrepeats HOK, RPS, and RB2. RPS is tandemly repeated while the other subrepeats occur once in each MRS. Chromosome 7, the smallest of the eight chromosomes, has been previously mapped. The complete physical map of this chromosome was used to analyze chromosome 7 diversity in six strains, including two well-characterized laboratory strains (1006 and WO-1) and four clinical ones. We found four types of events to explain the genomic diversity: 1) Chromosome length polymorphism (CLP) results from expansion and contraction of the RPS; 2) reciprocal translocation occurs at the MRS loci; 3) chromosomal deletion; and (4) trisomy of individual chromosomes. These four phenomena play an important role in generating genomic diversity in C. albicans.

Venturia inaequalis, the causal agent of apple scab, is controlled primarily by fungicides. Long-term, extensive fungicide use has led to the development of resistance to multiple fungicides. To assess fungicide resistance, isolates of V. inaequalis were collected from Indiana and Michigan orchards. Single-spore derived isolates were evaluated by mycelium growth assays with previously determined discriminatory doses on media containing dodine, kresoxim-methyl, myclobutanil, or thiophanate-methyl. Of 195 isolates tested, 5.2, 0.7, 57.0, and 92.6% of isolates were found to be resistant to dodine, kresoxim-methyl, myclobutanil, and thiophanate-methyl, respectively. This is the first report of kresoxim-methyl field resistance in these states. Isolates resistant or shifted to a single fungicide were often found to have multiple fungicide resistance. Of all isolates tested, 38% were identified as resistant or shifted to two fungicides, and 12% were resistant or shifted to all four fungicides tested. No fitness penalty was found for isolates resistant to multiple fungicides based on a statistical analysis of mycelial growth and conidial production.

Control strategies for Venturia inaequalis rely heavily on chemical fungicides. Single-site fungicides such as the quinone-outside inhibitors (QoI) have been used in Michigan apple orchards for more than 11 years. In 2008, we sampled eight commercial orchards in the Fruit Ridge growing region of Michigan in which apple scab control failures were observed on ‘McIntosh’ apple following applications of kresoxim-methyl or trifloxystrobin. QoI resistance was assessed in 210 total isolates (a total of 17 orchards) using a spore germination assay and in 319 isolates using a polymerase chain reaction (PCR) assay to detect the G143A mutation located within the V. inaequalis cytochrome b gene (CYTB). The G143A mutation is known to confer high-level QoI resistance in plant-pathogenic fungi. QoI resistance was confirmed in 50 and 64% of the isolates tested with the spore germination and PCR assays, respectively, and there was a 97% concordance observed between the assays. In 2009, we sampled and examined an additional 1,201 V. inaequalis isolates from 64 orchards in Michigan and 86 isolates from four baseline sites in Ohio. All of these isolates were assayed for the G143A mutation and it was detected within 67 and 0% of the Michigan and Ohio isolates, respectively. Our results indicate the widespread occurrence of QoI resistance in Michigan commercial orchard populations of V. inaequalis. Loss of QoI fungicides further limits the arsenal of fungicides available to commercial apple growers for successful scab management.

Parasexual genetic analysis of