Tetratheca erubescens is a narrowly endemic species including ~6300 plants restricted to a 2-km2 distribution on the south Koolyanobbing Range Banded Ironstone Formation (BIF) in Western Australia. A key objective of the present study was to characterise population genetic variation, and its spatial structuring across the entire distribution of T. erubescens, to enable a quantification of genetic variation that may be affected by proposed mining of the BIF. In total, 436 plants (~30 at each of 14 sites) from across the entire distribution were sampled, genotyped and scored for allelic variation at 11 polymorphic microsatellite loci. Fifty-nine alleles were detected (mean alleles per locus=5.36, range 2–10), and observed heterozygosity was low to moderate and typically lower than expected heterozygosity across all loci (mean observed heterozygosity (Ho)=0.41, mean expected heterozygosity (He)=0.48). Given the restricted distribution of T. erubescens, overall genetic structuring was surprisingly strong (overall FST=0.098). A range-wide spatial autocorrelation analysis indicated a significant positive genetic correlation at distances up to 450m, largely corresponding to the scale of more-or-less continuous distribution within each of two geographic clusters. In support, a STRUCTURE analysis identified an optimal number of genetic clusters as K=2, with assignment of individuals to one of two genetic clusters corresponding with the main geographic clusters. The genetic impact of proposed mining on T. erubescens was assessed on the basis of identifying plants within the proposed mine footprint (all plants from 4 of 14 sites). Repeating analyses of genetic variation after removal of these samples, and comparing to the complete dataset adjusted for sample size, resulted in the loss of one (very rare: overall frequency=0.001) allele (i.e. 58 of 59 alleles (98.3%) were recovered). All other parameters of genetic variation (mean Na, Ne, I, Ho, He, F) were unaffected. Consequently, although up to 22% of all plants fall within the mine footprint and, therefore, may be lost, <2% of alleles detected will be lost, and other genetic parameters remained unaffected. Although these results suggest that the proposed mining will result in a negligible impact on the assessed genetic variation and its spatial structuring in T. erubescens, further research on impacts to, and management of, quantitative genetic variation and key population genetic processes is required.
Multi-GBS: A massively multiplexed GBS-based protocol optimized for large, repetitive conifer genomes
