The discovery of microsatellite markers for 'Hardenbergia violacea' (Fabaceae), using next-generation sequencing

Abstract

'Hardenbergia violacea' is a climbing to prostrate shrub endemic to woodlands in the south east of Australia. We used next generation sequencing to develop a species-specific primer set. We obtained 106,529 reads in FASTA format with an average read length of 548 bp. DNA was successfully amplified for 20 markers and eight markers were characterized. The degree of polymorphism of these loci was tested on 95 samples from five populations. The total number of alleles ranged from two to 11 and observed and expected heterozygosities ranged from 0.0 to 0.807 and 0.0 to 0.933, respectively. Inbreeding coefficients were highly variable among populations, suggesting a mixture of selfing and outcrossing in nature. The characterized loci will be useful for studies of population structure, gene flow, mating systems, and the conservation and selection of provenances for restoration projects in degraded habitats.