Development of EST microsatellite markers for the Tasmanian palaeoendemic conifer Lagarostrobos franklinii (Hook. f.) Quinn (Podocarpaceae)

Abstract

Abstract Nuclear Expressed Sequence Tag (EST) microsatellite markers were developed for the Tasmanian palaeoendemic conifer Lagarostrobos franklinii (Hook.-f.) Quinn for genetic studies. RNAseq data was mined for EST microsatellites, and primer pairs were synthesised from 70 contigs with 50 producing amplification products. Of these 50, 10 reliably amplified and displayed polymorphism across 8 samples representing the entire species range. The genetic diversity of these 10 loci was then examined in three wild populations (84 samples). The number of alleles varied from two to thirteen per locus with the average number of alleles per population ranging between 3.0 – 4.7. Observed and expected heterozygosity ranged from 0.34 – 0.42 and 0.37 – 0.44, respectively. Marker cross-amplification was tested in the New Zealand sister species Manoao colensoi (Hook. f.) Molloy, but no markers amplified reliably, which possibly reflects the age of divergence between these species (~64 million years). These are the first microsatellite markers developed for the monotypic genus Lagarostrobos. They will be valuable for assessing the species extant genetic diversity, the impact of past climatic perturbations and human disturbance and the role of clonal propagation in recruitment.