Limited genetic diversity across pathogen populations responsible for the global emergence of boxwood blight identified using SSRs

Abstract

Boxwood blight is an emerging disease of ornamental as well as native boxwood. The disease became widely established in Europe at the beginning of the 21st century, prior to its more recent discovery in North America and Asia. Two sister-species of fungi cause the disease, Calonectria pseudonaviculata (Cps) and C. henricotiae (Che). Prior efforts to quantify intraspecific genetic polymorphisms of Cps and Che have yielded little information, limiting the ability to understand the evolution and migration of these pathogens. This study describes the development and implementation of simple sequence repeat (SSR) markers to analyse genetic diversity from a global collection of Cps and Che isolates, representing major blight outbreaks since the disease was first identified in the UK in the late 1990s. Analysis of the Cps CB002 genome sequence identified 180 single copy SSR loci using stringent search criteria, 11 of which were polymorphic and used to screen a global sample of 306 isolates. Fourteen multilocus genotypes of Cps and two multilocus genotypes of Che were identified. Twelve of the 14 Cps genotypes differed from each other by a single allele. The most common Cps genotype was found on all continents where boxwood blight is confirmed. Based on measurement of linkage disequilibrium, Cps showed no evidence of sexual recombination. Further in silico analysis identified 1594 SSRs using relaxed SSR definition criteria. Comparison of these SSR-containing loci with Cps and Che genome sequences representing three different genotypes demonstrated that single nucleotide polymorphisms might serve as informative genetic markers for future studies.