Pinus albicaulis (whitebark pine) is a widely-distributed but rapidly declining high elevation western North American tree and a candidate for listing under the U.S. Endangered Species Act. Our objectives were to develop reliable nuclear microsatellite markers that can be used to assess within-population genetic diversity as well as seed and pollen migration dynamics, and to validate markers using two geographically proximal P. albicaulis populations. We identified 1,667 microsatellite-containing sequences from shotgun DNA libraries of P. albicaulis. Primer pairs were designed for 308 unique microsatellite-containing loci, and these were evaluated for PCR amplification success and segregation in a panel of diploid needle tissue. DNA was extracted with an SDS protocol, and primers were screened through gel electrophoresis. Microsatellites were genotyped through fluorescent primer fragment analysis. Ten novel and 13 transferred loci were found to be reproducible in analyses based on 20 foliage samples from each of two locations: Henderson Mountain, Custer Gallatin National Forest, Montana, and Mt. Washburn, Yellowstone National Park, Wyoming (USA). Transferred loci had higher numbers of alleles and expected heterozygosities than novel loci, but also revealed evidence for a higher frequency of null alleles. Eight of the 13 transferred loci deviated significantly from Hardy-Weinberg Equilibrium, and showed large positive FIS values that were likely inflated by null alleles. Mantel’s tests of transferred and novel markers showed no correlation between genetic and geographic distances within or among the two sampled populations. AMOVA suggests that 91% of genetic variability occurs within populations and 9% between the two populations. Studies assessing genetic diversity using these microsatellite loci can help guide future management and restoration activities for P. albicaulis.
Eastern hemlock (Tsuga canadensis (L.) Carrière) is a shade-tolerant, slow-growing tree once common in forests across the Great Lakes region. It was heavily exploited in the late 19th and early 20th centuries and now experiences limited regeneration across much of its range. This failure to regenerate has been ascribed to poor seedbed conditions, insufficient canopy openings, warmer climate, and browsing by white-tailed deer (Odocoileus virginianus (Zimmermann, 1780)) or snowshoe hare (Lepus americanus Erxleben, 1777). To test whether deer or hare limit hemlock regeneration, we studied >2000 hemlock seedlings inside and adjacent to experimental deer exclosures at 59 sites randomly distributed across hemlock and hemlock-component stands in northern Wisconsin and the western Upper Peninsula of Michigan in the United States. We monitored local deer and hare abundance, seedling growth, and seedling survival for 20 years. Two First Nations reservations showed lower deer density and greater survival and growth of unprotected seedlings than three national forests of the United States. Cohorts of hemlock seedlings protected from deer survived at a rate four times higher than those exposed to deer (59.3% vs. 15.0%) and shared a combined height 5.2 times greater. Hare densities significantly affected seedling survival only within exclosures. This extensive, long-term study identifies deer as the primary factor constraining hemlock regeneration in the region.
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