Development of polymorphic microsatellite markers for the genetic characterisation of Knoxdaviesia proteae (Ascomycota: Microascales) using ISSR-PCR and pyrosequencing
Loading...
Date
Authors
Aylward, Janneke
Dreyer, Leanne Laurette
Steenkamp, Emma Theodora
Wingfield, Michael J.
Roets, Francois
Journal Title
Journal ISSN
Volume Title
Publisher
Springer
Abstract
Knoxdaviesia proteae is one of the first native
ophiostomatoid fungi discovered in South Africa, where it
consistently occurs in the infructescences of the iconic Cape
Biome plant, Protea repens. Although numerous studies have
been undertaken to better understand the ecology of
K. proteae, many questions remain to be answered, particularly
given its unique niche and association with arthropods
for dispersal.We describe the development and distribution of
microsatellite markers in K. proteae through Interspersed
Simple Sequence Repeat-Polymerase Chain Reaction (ISSR-PCR)
enrichment and pyrosequencing. A large proportion of
the 31492 sequences obtained from sequencing the enriched
genomic DNA were characterised by microsatellites
consisting of short tandem repeats and di- and tri-nucleotide
motifs. Seventeen percent of these microsatellites contained
flanking regions sufficient for primer design. Twenty-three
primer pairs were tested, of which 12 amplified and 10 generated polymorphic fragments in K. proteae. Half of these
could be transferred to the sister species, K. capensis. The
developedmarkers will be used to investigate the reproductive
system, genetic diversity and dispersal strategies of
K. proteae.
Description
Keywords
Knoxdaviesia, Microsatellites, Ophiostomatoid, Pyrosequencing, Interspersed Simple Sequence Repeat-Polymerase Chain Reaction (ISSR-PCR)
Sustainable Development Goals
Citation
Aylward, J, Dreyer, LL, Steenkamp, ET, Wingfield, MJ & Roets, F 2014, 'Development of polymorphic microsatellite markers for the genetic characterisation of Knoxdaviesia proteae (Ascomycota: Microascales) using ISSR-PCR and pyrosequencing', Mycological Progress, vol. 13. no. 2, pp. 439-444.