European Journal of Plant Pathology (2019) 155, 81-91

C. Cornejo, B. Sever, Q. Kupper, S. Prospero and D. Rigling (2019)
A multiplexed genotyping assay to determine vegetative incompatibility and mating type in Cryphonectria parasitica
European Journal of Plant Pathology 155 (1), 81-91
Abstract: The highly destructive chestnut blight disease can be successfully controlled by infecting the virulent strain of C. parasitica with the hyperparasitic mycovirus CHV-1. The artificial application of the virus-induced hypovirulence, however, requires that the vegetative compatibility (vc) diversity in the target C. parasitica population be determined beforehand. Conventional vc type determination by pairing the unknown isolate with tester strains of known vc types is time consuming and not always reliable. A genotyping assay was developed that uses DNA from C. parasitica isolates and discriminates the six known di-allelic vic loci and the two mating idiomorphs in two fluorescence-labeled multiplex PCRs. Validation was conducted by successfully genotyping the 74 known European vc tester strains and a conventionally characterized natural population of C. parasitica. Cross-species amplification tests with three Cryphonectria species (C. japonica, C. naterciae, and C. radicalis) revealed a high amplification specificity for C. parasitica, while the analytical sensitivity of the assay was established at 20 pg per reaction of genomic DNA. In conclusion, this fluorescent multiplex genotyping assay offers a simple and high-throughput tool for the characterization of the vegetative compatibility and mating type of C. parasitica at population level, which is particularly important for the application of mycovirus-mediated biological control of chestnut blight.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Link to article at publishers website
Database assignments for author(s): Daniel Rigling

Research topic(s) for pests/diseases/weeds:
general biology - morphology - evolution
biocontrol - natural enemies
Research topic(s) for beneficials or antagonists:
general biology - morphology - evolution

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Cryphonectria parasitica
Cryphonectria parasitica hypoviruses (antagonist)	Cryphonectria parasitica