Multilocus PCR Assays Elucidate Vegetative Incompatibility Gene Profiles of Cryphonectria parasitica in the United States
Short DP, Double M, Nuss DL, Stauder CM, MacDonald W, Kasson MT. 2015. Multilocus PCR Assays Elucidate Vegetative Incompatibility Gene Profiles of Cryphonectria parasitica in the United States. Applied and Environmental Microbiology
Chestnut blight is a devastating disease of Castanea spp. Mycoviruses that reduce virulence (hypovirulence) of the causative agent, Cryphonectria parasitica, can be used to manage chestnut blight. However, vegetative incompatibility (vic) barriers that restrict anastomosis-mediated virus transmission hamper hypovirulence efficacy. In order to effectively determine the vegetative incompatibility genetic structure of C. parasitica field populations, we have designed PCR primer sets that selectively amplify and distinguish alleles for each of the six known di-allelic C. parasitica vic genetic loci. PCR assay results were validated using a panel of 64 European tester strains with genetically determined vic genotypes. Analysis of 116 C. parasitica isolates collected from five locations in the eastern U.S. revealed 39 unique vic genotypes and generally good agreement between PCR and tester strain co-culturing assays in terms of vic diversity and genotyping. However, incongruences were observed for isolates from multiple locations and suggested that the co-culturing assay can overestimate diversity at the six known vic loci. The availability of molecular tools for rapid and precise vic genotyping significantly improves the ability to predict and evaluate the efficacy of hypovirulence and related management strategies.
- Type
- Journal Article
- Authors
- Short, Dylan; Double, Mark; Nuss, Donald; Stauder, Cameron; MacDonald, William; Kasson, Matthew
- Units
- GRSM , SHEN
- Keywords
- APHN, Appalachian Highlands Network, chestnut blight, Cryphonectria parasitica, Great Smoky Mountains National Park, GRSM, GRSM-01039, hypovirus transmission, Population Biology, SER, Southeast Region, vic loci