A Multiplex PCR Approach to Determine Vegetative Incompatibility Genotypes and Mating Type in Cryphonectria parasitica

Quirin Kupper; Carolina Cornejo

doi:10.1007/978-1-0716-2517-0_25

A Multiplex PCR Approach to Determine Vegetative Incompatibility Genotypes and Mating Type in Cryphonectria parasitica

Methods Mol Biol. 2022:2536:435-446. doi: 10.1007/978-1-0716-2517-0_25.

Authors

Quirin Kupper¹, Carolina Cornejo²

Affiliations

¹ Swiss Federal Research Institute WSL, Birmensdorf, Switzerland.
² Swiss Federal Research Institute WSL, Birmensdorf, Switzerland. carolina.cornejo@wsl.ch.

PMID: 35819619
DOI: 10.1007/978-1-0716-2517-0_25

Abstract

This chapter presents a genotyping assay that uses DNA isolated from axenic cultures of Cryphonectria parasitica, which discriminates the six known diallelic vic loci and the two mating idiomorphs (MAT gene) based on (i) modified primer, labeled with a fluorescent dye, (ii) multiplex polymerase chain reaction (multiplex-PCR), and (iii) capillary electrophoresis system. Alternatively, we show that the same primer set is suitable for conventional PCR of each vic locus and MAT gene using nonmodified primer and agarose gel electrophoresis.

Keywords: Biocontrol; Chestnut blight; Diagnostics; Duplex-PCR; Fluorochrome; Genetic diversity; Multiplex-PCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Ascomycota* / genetics
Genotype
Multiplex Polymerase Chain Reaction*
Reproduction / genetics

Supplementary concepts

Cryphonectria parasitica