Development of polymorphic microsatellite markers issued from pyrosequencing technology for the medicinal mushroom Agaricus subrufescens

Abstract

The recently described procedure of microsatellite-enriched library pyrosequencing was used to isolate microsatellite loci in the gourmet and medicinal mushroom Agaricus subrufescens. Three hundred and five candidate loci containing at least one simple sequence repeats (SSR) locus and for which primers design was successful, were obtained. From a subset of 95 loci, 35 operational and polymorphic SSR markers were developed and characterized on a sample of 14 A. subrufescens genotypes from diverse origins. These SubSSR markers each displayed from two to 10 alleles with an average of 4.66 alleles per locus. The observed heterozygosity ranged from 0 to 0.71. Several multiplex combinations can be set up, making it possible to genotype up to six markers easily and simultaneously. Cross-amplification in some closely congeneric species was successful for a subset of loci. The 35 microsatellite markers developed here provide a highly valuable molecular tool to study genetic diversity and reproductive biology of A. subrufescens.

The Genetic Linkage Map of the Medicinal Mushroom Agaricus subrufescens Reveals Highly Conserved Macrosynteny with the Congeneric Species Agaricus bisporus

“…For each SSI, the level of ploidy ( n + n vs. n ) was determined with a multilocus genotype test based on 21 genomic microsatellite markers (hereinafter referred to as SubSSR for A . Sub rufescens simple sequence repeats) ( Foulongne-Oriol et al 2012c ) that showed polymorphism between the two parental homokaryons: SSIs that did not exhibit heteromorphic profile at any loci were considered homokaryotic ( Thongklang et al 2014a ). Among the 380 SSIs tested, 79 were found homokaryotic and were thus selected for further mapping purposes.…”

“…The genotyping of genomic SubSSR markers ( Foulongne-Oriol et al 2012c ) followed from the homokaryon selection test described above. In addition to this first set of 21 microsatellite markers, EST-SSRs (hereinafter referred to as ES-SSR) were also used for mapping ( Foulongne-Oriol et al 2014 ).…”

“…In addition to this first set of 21 microsatellite markers, EST-SSRs (hereinafter referred to as ES-SSR) were also used for mapping ( Foulongne-Oriol et al 2014 ). Amplification conditions, electrophoretic separation, and visualization were performed according to Foulongne-Oriol et al (2012c) . All microsatellite primers used for mapping are listed in Supplemental Material, Table A in File S1 .…”

“…Breeding work on this species is in its infancy ( Kerrigan and Wach 2008 ; Llarena-Hernandez et al 2013 ) due to several bottlenecks, including a lack of knowledge on its ecology, reproductive biology, biodiversity, and genetics ( Largeteau et al 2011 ). The recent development of molecular markers and available expressed sequence tag (EST) resources have enriched our toolbox for studying the biology of this mushroom ( Foulongne-Oriol et al 2012c , 2014 ). The clarification of its amphithallic life cycle, and of the interfertility between field specimens from different continents ( Rocha de Brito et al 2016 ; Thongklang et al 2014a ), provide a sound basis to exploit the genetic diversity of this species, and to conduct controlled mating of selected genotypes.…”

Do spawn storage conditions influence the colonization capacity of a wheat-straw-based substrate by Agaricus subrufescens?

“…CA646 and CA647 are two cultivars sold in Europe by Mycelia (Belgium) under the reference numbers 7700 and 7703, respectively. CA560 was isolated in Botucatu, Brazil (1999) from a commercial farm and CA572 is a Brazilian cultivar purchased by D.C. Zied in 2007; both strains have proved to be genetically different [9]. Their original reference numbers are ABL-99/28 and ABL-07/58, respectively, in the Culture Collection of the Edible and Medicinal activé es aprè s stockage en comparaison avec le contrôle ainsi que la transformation du substrat, comme dé crit par les ratios d'aromaticité et d'humification calculé s à partir des donné es de RMN.…”