North American tree migration paced by climate in the West, lagging in the East.

Tree fecundity and recruitment have not yet been quantified at scales needed to anticipate biogeographic shifts in response to climate change. By separating their responses, this study shows coherence across species and communities, offering the strongest support to date that migration is in progress with regional limitations on rates. The southeastern continent emerges as a fecundity hotspot, but it is situated south of population centers where high seed production could contribute to poleward population spread. By contrast, seedling success is highest in the West and North, serving to partially offset limited seed production near poleward frontiers. The evidence of fecundity and recruitment control on tree migration can inform conservation planning for the expected long-term disequilibrium between climate and forest distribution.

Future carbon sequestration potential in a widespread transcontinental boreal tree species: Standing genetic variation matters!

Climate change (CC) necessitates reforestation/afforestation programs to mitigate its impacts and maximize carbon sequestration. But comprehending how tree growth, a proxy for fitness and resilience, responds to CC is critical to maximize these programs' effectiveness. Variability in tree response to CC across populations can notably be influenced by the standing genetic variation encompassing both neutral and adaptive genetic diversity. Here, a framework is proposed to assess tree growth potential at the population scale while accounting for standing genetic variation. We applied this framework to black spruce (BS, Picea mariana [Mill] B.S.P.), with the objectives to (1) determine the key climate variables having impacted BS growth response from 1974 to 2019, (2) examine the relative roles of local adaptation and the phylogeographic structure in this response, and (3) project BS growth under two Shared Socioeconomic Pathways while taking standing genetic variation into account. We modeled growth using a machine learning algorithm trained with dendroecological and genetic data obtained from over 2600 trees (62 populations divided in three genetic clusters) in four 48-year-old common gardens, and simulated growth until year 2100 at the common garden locations. Our study revealed that high summer and autumn temperatures negatively impacted BS growth. As a consequence of warming, this species is projected to experience a decline in growth by the end of the century, suggesting maladaptation to anticipated CC and a potential threat to its carbon sequestration capacity. This being said, we observed a clear difference in response to CC within and among genetic clusters, with the western cluster being more impacted than the central and eastern clusters. Our results show that intraspecific genetic variation, notably associated with the phylogeographic structure, must be considered when estimating the response of widespread species to CC.

Increasing fire and the decline of fire adapted black spruce in the boreal forest.

Intensifying wildfire activity and climate change can drive rapid forest compositional shifts. In boreal North America, black spruce shapes forest flammability and depends on fire for regeneration. This relationship has helped black spruce maintain its dominance through much of the Holocene. However, with climate change and more frequent and severe fires, shifts away from black spruce dominance to broadleaf or pine species are emerging, with implications for ecosystem functions including carbon sequestration, water and energy fluxes, and wildlife habitat. Here, we predict that such reductions in black spruce after fire may already be widespread given current trends in climate and fire. To test this, we synthesize data from 1,538 field sites across boreal North America to evaluate compositional changes in tree species following 58 recent fires (1989 to 2014). While black spruce was resilient following most fires (62%), loss of resilience was common, and spruce regeneration failed completely in 18% of 1,140 black spruce sites. In contrast, postfire regeneration never failed in forests dominated by jack pine, which also possesses an aerial seed bank, or broad-leaved trees. More complete combustion of the soil organic layer, which often occurs in better-drained landscape positions and in dryer duff, promoted compositional changes throughout boreal North America. Forests in western North America, however, were more vulnerable to change due to greater long-term climate moisture deficits. While we find considerable remaining resilience in black spruce forests, predicted increases in climate moisture deficits and fire activity will erode this resilience, pushing the system toward a tipping point that has not been crossed in several thousand years.

Is there tree senescence? The fecundity evidence.

Despite its importance for forest regeneration, food webs, and human economies, changes in tree fecundity with tree size and age remain largely unknown. The allometric increase with tree diameter assumed in ecological models would substantially overestimate seed contributions from large trees if fecundity eventually declines with size. Current estimates are dominated by overrepresentation of small trees in regression models. We combined global fecundity data, including a substantial representation of large trees. We compared size-fecundity relationships against traditional allometric scaling with diameter and two models based on crown architecture. All allometric models fail to describe the declining rate of increase in fecundity with diameter found for 80% of 597 species in our analysis. The strong evidence of declining fecundity, beyond what can be explained by crown architectural change, is consistent with physiological decline. A downward revision of projected fecundity of large trees can improve the next generation of forest dynamic models.

Rooting depth and xylem vulnerability are independent woody plant traits jointly selected by aridity, seasonality, and water table depth.

Evolutionary radiations of woody taxa within arid environments were made possible by multiple trait innovations including deep roots and embolism-resistant xylem, but little is known about how these traits have coevolved across the phylogeny of woody plants or how they jointly influence the distribution of species. We synthesized global trait and vegetation plot datasets to examine how rooting depth and xylem vulnerability across 188 woody plant species interact with aridity, precipitation seasonality, and water table depth to influence species occurrence probabilities across all biomes. Xylem resistance to embolism and rooting depth are independent woody plant traits that do not exhibit an interspecific trade-off. Resistant xylem and deep roots increase occurrence probabilities in arid, seasonal climates over deep water tables. Resistant xylem and shallow roots increase occurrence probabilities in arid, nonseasonal climates over deep water tables. Vulnerable xylem and deep roots increase occurrence probabilities in arid, nonseasonal climates over shallow water tables. Lastly, vulnerable xylem and shallow roots increase occurrence probabilities in humid climates. Each combination of trait values optimizes occurrence probabilities in unique environmental conditions. Responses of deeply rooted vegetation may be buffered if evaporative demand changes faster than water table depth under climate change.

A critical thermal transition driving spring phenology of Northern Hemisphere conifers.

Despite growing interest in predicting plant phenological shifts, advanced spring phenology by global climate change remains debated. Evidence documenting either small or large advancement of spring phenology to rising temperature over the spatio-temporal scales implies a potential existence of a thermal threshold in the responses of forests to global warming. We collected a unique data set of xylem cell-wall-thickening onset dates in 20 coniferous species covering a broad mean annual temperature (MAT) gradient (-3.05 to 22.9°C) across the Northern Hemisphere (latitudes 23°-66° N). Along the MAT gradient, we identified a threshold temperature (using segmented regression) of 4.9 ± 1.1°C, above which the response of xylem phenology to rising temperatures significantly decline. This threshold separates the Northern Hemisphere conifers into cold and warm thermal niches, with MAT and spring forcing being the primary drivers for the onset dates (estimated by linear and Bayesian mixed-effect models), respectively. The identified thermal threshold should be integrated into the Earth-System-Models for a better understanding of spring phenology in response to global warming and an improved prediction of global climate-carbon feedbacks.

Photoperiod and temperature as dominant environmental drivers triggering secondary growth resumption in Northern Hemisphere conifers.

Wood formation consumes around 15% of the anthropogenic CO2 emissions per year and plays a critical role in long-term sequestration of carbon on Earth. However, the exogenous factors driving wood formation onset and the underlying cellular mechanisms are still poorly understood and quantified, and this hampers an effective assessment of terrestrial forest productivity and carbon budget under global warming. Here, we used an extensive collection of unique datasets of weekly xylem tissue formation (wood formation) from 21 coniferous species across the Northern Hemisphere (latitudes 23 to 67°N) to present a quantitative demonstration that the onset of wood formation in Northern Hemisphere conifers is primarily driven by photoperiod and mean annual temperature (MAT), and only secondarily by spring forcing, winter chilling, and moisture availability. Photoperiod interacts with MAT and plays the dominant role in regulating the onset of secondary meristem growth, contrary to its as-yet-unquantified role in affecting the springtime phenology of primary meristems. The unique relationships between exogenous factors and wood formation could help to predict how forest ecosystems respond and adapt to climate warming and could provide a better understanding of the feedback occurring between vegetation and climate that is mediated by phenology. Our study quantifies the role of major environmental drivers for incorporation into state-of-the-art Earth system models (ESMs), thereby providing an improved assessment of long-term and high-resolution observations of biogeochemical cycles across terrestrial biomes.

Dominance of coniferous and broadleaved trees drives bacterial associations with boreal feather mosses.

The composition of ecologically important moss-associated bacterial communities seems to be mainly driven by host species but may also be shaped by environmental conditions related with tree dominance. The moss phyllosphere has been studied in coniferous forests while broadleaf forests remain understudied. To determine if host species or environmental conditions defined by tree dominance drives the bacterial diversity in the moss phyllosphere, we used 16S rRNA gene amplicon sequencing to quantify changes in bacterial communities as a function of host species (Pleurozium schreberi and Ptilium crista-castrensis) and forest type (coniferous black spruce versus deciduous broadleaf trembling aspen) in eastern Canada. The overall composition of moss phyllosphere was defined by the interaction of both factors, though most of the bacterial phyla were determined by a strong effect of forest type. Bacterial α-diversity was highest in spruce forests, while there was greater turnover (ß-diversity) and higher γ-diversity in aspen forests. Unexpectedly, Cyanobacteria were much more relatively abundant in aspen than in spruce forests, with the cyanobacteria family Nostocaceae differing the most between forest types. Our results advance the understanding of moss-associated microbial communities among coniferous and broadleaf deciduous forests, which are important with the increasing changes in tree dominance in the boreal system.

Site index as a predictor of the effect of climate warming on boreal tree growth.

The boreal forest represents the terrestrial biome most heavily affected by climate change. However, no consensus exists regarding the impacts of these changes on the growth of tree species therein. Moreover, assessments of young tree responses in metrics transposable to forest management remain scarce. Here, we assessed the impacts of climate change on black spruce (Picea mariana [Miller] BSP) and jack pine (Pinus banksiana Lambert) growth, two dominant tree species in boreal forests of North America. Starting with a retrospective analysis including data from 2591 black spruces and 890 jack pines, we forecasted trends in 30-year height growth at the transitions from closed to open boreal coniferous forests in Québec, Canada. We considered three variables: (1) height growth, rarely used, but better-reflecting site potential than other growth proxies, (2) climate normals corresponding to the growth period of each stem, and (3) site type (as a function of texture, stoniness, and drainage), which can modify the effects of climate on tree growth. We found a positive effect of vapor pressure deficit on the growth of both species, although the effect on black spruce leveled off. For black spruce, temperatures had a positive effect on the height at 30 years, which was attenuated when and where climatic conditions became drier. Conversely, drought had a positive effect on height under cold conditions and a negative effect under warm conditions. Spruce growth was also better on mesic than on rocky and sub-hydric sites. For portions of the study areas with projected future climate within the calibration range, median height-change varied from 10 to 31% for black spruce and from 5 to 31% for jack pine, depending on the period and climate scenario. As projected increases are relatively small, they may not be sufficient to compensate for potential increases in future disturbances like forest fires.

North America's oldest boreal trees are more efficient water users due to increased [CO2], but do not grow faster.

Due to anthropogenic emissions and changes in land use, trees are now exposed to atmospheric levels of [[Formula: see text]] that are unprecedented for 650,000 y [Lüthi et al. (2008) Nature 453:379-382] (thousands of tree generations). Trees are expected to acclimate by modulating leaf-gas exchanges and alter water use efficiency which may result in forest productivity changes. Here, we present evidence of one of the strongest, nonlinear, and unequivocal postindustrial increases in intrinsic water use efficiency ([Formula: see text]) ever documented (+59%). A dual-isotope tree-ring analysis ([Formula: see text] and [Formula: see text]) covering 715 y of growth of North America's oldest boreal trees (Thuja occidentalis L.) revealed an unprecedented increase in [Formula: see text] that was directly linked to elevated assimilation rates of [Formula: see text] (A). However, limited nutrient availability, changes in carbon allocation strategies, and changes in stomatal density may have offset stem growth benefits awarded by the increased [Formula: see text] Our results demonstrate that even in scenarios where a positive [Formula: see text] fertilization effect is observed, other mechanisms may prevent trees from assimilating and storing supplementary anthropogenic emissions as above-ground biomass. In such cases, the sink capacity of forests in response to changing atmospheric conditions might be overestimated.

Growing-season frost is a better predictor of tree growth than mean annual temperature in boreal mixedwood forest plantations.

Increase in frost damage to trees due to earlier spring dehardening could outweigh the expected increase in forest productivity caused by climate warming. We quantified the impact of growing-season frosts on the performance of three spruce species (white, black, and Norway spruce) and various seed sources with different frost tolerance in two plantations, established on both sides of the eastern Canadian boreal-temperate forest ecotone. The objectives of this study were to determine (a) if spruce species and seed sources planted in sites far from their natural provenance would be less adapted to local site conditions, leading to increased frost damage and reduced height growth; (b) at which height above the ground growing-season frosts ceased to damage apical meristems; and (c) if height growth was best predicted by extreme climatic events (growing-season frosts) or by mean annual or summer temperature. At each site and for all spruce species and seed sources, we cross-sectioned spruce trees at different heights above the ground. Tree rings were cross-dated and screened for frost rings, which were then given a severity score based on cellular damage. Frost severity reduced height growth of all spruce species and provenances at both sites. Height growth of the non-native Norway spruce was the most reduced by frost severity and was the smallest species at both sites. Frost caused the highest growth reduction in white spruce at the boreal mixedwood site and had the least effect on black spruce at both sites. For all spruce species, height growth was affected up to 2 m above the ground. Model selection based on corrected Akaike's information criteria (AICc) identified that minimum temperature in May was by far the best climate variable predicting tree growth (AICc weight = 1), highlighting the importance of considering extreme climatic events, which are likely to increase in the future.

Strong overestimation of water-use efficiency responses to rising CO2 in tree-ring studies.

The carbon isotope ratio (δ13 C) in tree rings is commonly used to derive estimates of the assimilation-to-stomatal conductance rate of trees, that is, intrinsic water-use efficiency (iWUE). Recent studies have observed increased iWUE in response to rising atmospheric CO2 concentrations (Ca ), in many different species, genera and biomes. However, increasing rates of iWUE vary widely from one study to another, likely because numerous covarying factors are involved. Here, we quantified changes in iWUE of two widely distributed boreal conifers using tree samples from a forest inventory network that were collected across a wide range of growing conditions (assessed using the site index, SI), developmental stages and stand histories. Using tree-ring isotopes analysis, we assessed the magnitude of increase in iWUE after accounting for the effects of tree size, stand age, nitrogen deposition, climate and SI. We also estimated how growth conditions have modulated tree physiological responses to rising Ca . We found that increases in tree size and stand age greatly influenced iWUE. The effect of Ca on iWUE was strongly reduced after accounting for these two variables. iWUE increased in response to Ca , mostly in trees growing on fertile stands, whereas iWUE remained almost unchanged on poor sites. Our results suggest that past studies could have overestimated the CO2 effect on iWUE, potentially leading to biased inferences about the future net carbon balance of the boreal forest. We also observed that this CO2 effect is weakening, which could affect the future capacity of trees to resist and recover from drought episodes.

Geographic isolation and climatic variability contribute to genetic differentiation in fragmented populations of the long-lived subalpine conifer Pinus cembra L. in the western Alps.

BACKGROUND: Genetic processes shape the modern-day distribution of genetic variation within and between populations and can provide important insights into the underlying mechanisms of evolution. The resulting genetic variation is often unequally partitioned within species' distribution range and especially large differences can manifest at the range limit, where population fragmentation and isolation play a crucial role in species survival. Despite several molecular studies investigating the genetic diversity and differentiation of European Alpine mountain forests, the climatic and demographic constrains which influence the genetic processes are often unknown. Here, we apply non-coding microsatellite markers to evaluate the sporadic peripheral and continuous populations of cembra pine (Pinus cembra L.), a long-lived conifer species that inhabits the subalpine treeline ecotone in the western Alps to investigate how the genetic processes contribute to the modern-day spatial distribution. Moreover, we corroborate our findings with paleoecological records, micro and macro-remains, to infer the species' possible glacial refugia and expansion scenarios. RESULTS: Four genetically distinct groups were identified, with Bayesian and FST based approaches, across the range of the species, situated in the northern, inner and south-western Alps. We found that genetic differentiation is substantially higher in marginal populations than at the center of the range, and marginal stands are characterized by geographic and genetic isolation due to spatial segregation and restricted gene flow. Moreover, multiple matrix regression approaches revealed effects of climatic heterogeneity in species' spatial genetic pattern. Also, population stability tests indicated that all populations had experienced a severe historical bottleneck, no heterozygosity excess was detected, suggesting that more recently population sizes have remained relatively stable. CONCLUSIONS: Our study demonstrated that cembra pine might have survived in multiple glacial refugia and subsequently recolonized the Alps by different routes. Modern-day marginal populations, at the edge of the species' range, could maintain stable sizes over long periods without inbreeding depression and preserve high amounts of genetic variation. Moreover, our analyses indicate that climatic variability has played a major role in shaping differentiation, in addition to past historical events such as migration and demographic changes.

Taxonomy, together with ontogeny and growing conditions, drives needleleaf species' sensitivity to climate in boreal North America.

Currently, there is no consensus regarding the way that changes in climate will affect boreal forest growth, where warming is occurring faster than in other biomes. Some studies suggest negative effects due to drought-induced stresses, while others provide evidence of increased growth rates due to a longer growing season. Studies focusing on the effects of environmental conditions on growth-climate relationships are usually limited to small sampling areas that do not encompass the full range of environmental conditions; therefore, they only provide a limited understanding of the processes at play. Here, we studied how environmental conditions and ontogeny modulated growth trends and growth-climate relationships of black spruce (Picea mariana) and jack pine (Pinus banksiana) using an extensive dataset from a forest inventory network. We quantified the long-term growth trends at the stand scale, based on analysis of the absolutely dated ring-width measurements of 2,266 trees. We assessed the relationship between annual growth rates and seasonal climate variables and evaluated the effects of various explanatory variables on long-term growth trends and growth-climate relationships. Both growth trends and growth-climate relationships were species-specific and spatially heterogeneous. While the growth of jack pine barely increased during the study period, we observed a growth decline for black spruce which was more pronounced for older stands. This decline was likely due to a negative balance between direct growth gains induced by improved photosynthesis during hotter-than-average growing conditions in early summers and the loss of growth occurring the following year due to the indirect effects of late-summer heat waves on accumulation of carbon reserves. For stands at the high end of our elevational gradient, frost damage during milder-than-average springs could act as an additional growth stressor. Competition and soil conditions also modified climate sensitivity, which suggests that effects of climate change will be highly heterogeneous across the boreal biome.

Long-term changes in the impacts of global warming on leaf phenology of four temperate tree species.

Contrary to the generally advanced spring leaf unfolding under global warming, the effects of the climate warming on autumn leaf senescence are highly variable with advanced, delayed, and unchanged patterns being all reported. Using one million records of leaf phenology from four dominant temperate species in Europe, we investigated the temperature sensitivities of spring leaf unfolding and autumn leaf senescence (ST , advanced or delayed days per degree Celsius). The ST of spring phenology in all of the four examined species showed an increase and decrease during 1951-1980 and 1981-2013, respectively. The decrease in the ST during 1981-2013 appears to be caused by reduced accumulation of chilling units. As with spring phenology, the ST of leaf senescence of early successional and exotic species started to decline since 1980. In contrast, for late successional species, the ST of autumn senescence showed an increase for the entire study period from 1951 to 2013. Moreover, the impacts of rising temperature associated with global warming on spring leaf unfolding were stronger than those on autumn leaf senescence. The timing of leaf senescence was positively correlated with the timing of leaf unfolding during 1951-1980. However, as climate warming continued, the differences in the responses between spring and autumn phenology gradually increased, so that the correlation was no more significant during 1981-2013. Our results further suggest that since 2000, due to the decreased temperature sensitivity of leaf unfolding the length of the growing season has not increased any more. These finding needs to be addressed in vegetation models used for assessing the effects of climate change.

Drivers of postfire soil organic carbon accumulation in the boreal forest.

The accumulation of soil carbon (C) is regulated by a complex interplay between abiotic and biotic factors. Our study aimed to identify the main drivers of soil C accumulation in the boreal forest of eastern North America. Ecosystem C pools were measured in 72 sites of fire origin that burned 2-314 years ago over a vast region with a range of ∆ mean annual temperature of 3°C and one of ∆ 500 mm total precipitation. We used a set of multivariate a priori causal hypotheses to test the influence of time since fire (TSF), climate, soil physico-chemistry and bryophyte dominance on forest soil organic C accumulation. Integrating the direct and indirect effects among abiotic and biotic variables explained as much as 50% of the full model variability. The main direct drivers of soil C stocks were: TSF >bryophyte dominance of the FH layer and metal oxide content >pH of the mineral soil. Only climate parameters related to water availability contributed significantly to explaining soil C stock variation. Importantly, climate was found to affect FH layer and mineral soil C stocks indirectly through its effects on bryophyte dominance and organo-metal complexation, respectively. Soil texture had no influence on soil C stocks. Soil C stocks increased both in the FH layer and mineral soil with TSF and this effect was linked to a decrease in pH with TSF in mineral soil. TSF thus appears to be an important factor of soil development and of C sequestration in mineral soil through its influence on soil chemistry. Overall, this work highlights that integrating the complex interplay between the main drivers of soil C stocks into mechanistic models of C dynamics could improve our ability to assess C stocks and better anticipate the response of the boreal forest to global change.

Spring phenology at different altitudes is becoming more uniform under global warming in Europe.

Under current global warming, high-elevation regions are expected to experience faster warming than low-elevation regions. However, due to the lack of studies based on long-term large-scale data, the relationship between tree spring phenology and the elevation-dependent warming is unclear. Using 652k records of leaf unfolding of five temperate tree species monitored during 1951-2013 in situ in Europe, we discovered a nonlinear trend in the altitudinal sensitivity (SA , shifted days per 100 m in altitude) in spring phenology. A delayed leaf unfolding (2.7 ± 0.6 days per decade) was observed at high elevations possibly due to decreased spring forcing between 1951 and 1980. The delayed leaf unfolding at high-elevation regions was companied by a simultaneous advancing of leaf unfolding at low elevations. These divergent trends contributed to a significant increase in the SA (0.36 ± 0.07 days 100/m per decade) during 1951-1980. Since 1980, the SA started to decline with a rate of -0.32 ± 0.07 days 100/m per decade, possibly due to reduced chilling at low elevations and improved efficiency of spring forcing in advancing the leaf unfolding at high elevations, the latter being caused by increased chilling. Our results suggest that due to both different temperature changes at the different altitudes, and the different tree responses to these changes, the tree phenology has shifted at different rates leading to a more uniform phenology at different altitudes during recent decades.

Does time since fire drive live aboveground biomass and stand structure in low fire activity boreal forests? Impacts on their management.

Boreal forests subject to low fire activity are complex ecosystems in terms of structure and dynamics. They have a high ecological value as they contain important proportions of old forests that play a crucial role in preserving biodiversity and ecological functions. They also sequester important amounts of carbon at the landscape level. However, the role of time since fire in controlling the different processes and attributes of those forests is still poorly understood. The Romaine River area experiences a fire regime characterized by very rare but large fires and has recently been opened to economic development for energy and timber production. In this study, we aimed to characterize this region in terms of live aboveground biomass, merchantable volume, stand structure and composition, and to establish relations between these attributes and the time since the last fire. Mean live aboveground biomass and merchantable volume showed values similar to those of commercial boreal coniferous forests. They were both found to increase up to around 150 years after a fire before declining. However, no significant relation was found between time since fire and stand structure and composition. Instead, they seemed to mostly depend on stand productivity and non-fire disturbances. At the landscape level, this region contains large amounts of biomass and carbon stored resulting from the long fire cycles it experiences. Although in terms of merchantable volume these forests seemed profitable for the forest industry, a large proportion were old forests or presented structures of old forests. Therefore, if forest management was to be undertaken in this region, particular attention should be given to these old forests in order to protect biodiversity and ecological functions. Partial cutting with variable levels of retention would be an appropriate management strategy as it reproduces the structural complexity of old forests.

Fine-scale assessment of genetic diversity of trembling aspen in northwestern North America.

BACKGROUND: In North America, the last ice age is the most recent event with severe consequences on boreal species' ranges. Phylogeographic patterns of range expansion in trembling aspen (Populus tremuloides) suggested that Beringia is likely to be a refugium and the "ice-free corridor" in Alberta may represent a region where small populations persisted during the last glacial maximum (LGM). The purpose of this study was to ascertain whether the origins of trembling aspen in western North America are reflected in the patterns of neutral genetic diversity and population structure. A total of 28 sites were sampled covering the northwestern part of aspen's distribution, from Saskatchewan to Alaska. Twelve microsatellite markers were used to describe patterns of genetic diversity. The genetic structure of trembling aspen populations was assessed by using multivariate analyses, Mantel correlograms, neighbor-joining trees and Bayesian analysis. RESULTS: Microsatellite markers revealed little to no neutral genetic structure of P. tremuloides populations in northwestern North America. Low differentiation among populations and small isolation by distance (IBD) were observed. The most probable number of clusters detected by STRUCTURE was K = 3 (∆K = 5.9). The individuals in the populations of the 3 clusters share a common gene pool and showed a high level of admixture. No evidence was found that either Beringia or the "ice-free corridor" were refugia. Highest allelic richness (AR) and lowest heterozygosity (Ho) were observed in Alberta foothills of the Rocky Mountains. CONCLUSIONS: Contrary to our hypothesis, our results showed that microsatellite markers revealed little to no genetic structure in P. tremuloides populations. Consequently, no divergent populations were observed near supposed refugia. The lack of detectable refugia in Beringia and in the "ice-free corridor" was due to high levels of gene flow between trembling apsen populations. More favorable environmental conditions for sexual reproduction and successful trembling aspen seedling establishment may have contributed to increase allelic richness through recombination in populations from the Albertan foothills of the Rocky Mountains.