Incidence of Venturia shoot blight in aspen (Populus tremuloides Michx.) varies with tree chemistry and genotype

Quaking aspen (Populus tremuloides) is a foundation tree species in North American forests, as well as a valuable source of wood and paper products. Quaking aspen exhibits substantial genetic variation within and among natural populations in phytochemical compounds that influence both interactions with herbivores and ecosystem dynamics. The potential association of these phytochemicals with disease resistance, however, is unknown. Here we present the results of a “natural experiment” in a common garden of quaking aspen genotypes infected with shoot blight (Venturia moreletii). We found that the incidence of shoot blight varied by 10-fold among aspen genotypes, and was strongly and negatively correlated with constitutive foliar concentrations of condensed tannins. Selection factors that shape the genetic and phytochemical architecture of aspen populations may thus simultaneously influence aspen resistance to pathogen attack, with consequences for individual tree fitness as well as community organization, via “extended phenotype” effects.     

Evidence for widespread selection in shaping the genomic landscape during speciation of Populus

Increasing our understanding of how evolutionary processes drive the genomic landscape of variation is fundamental to a better understanding of the genomic consequences of speciation. However, genome‐wide patterns of within‐ and between‐ species variation have not been fully investigated in most forest tree species despite their global ecological and economic importance. Here, we use whole‐genome resequencing data from four Populus species spanning the speciation continuum to reconstruct their demographic histories and investigate patterns of diversity and divergence within and between species. Using Populus trichocarpa as an outgroup species, we further infer the genealogical relationships and estimate the extent of ancient introgression among the three aspen species (Populus tremula, Populus davidiana and Populus tremuloides) throughout the genome. Our results show substantial variation in these patterns along the genomes with this variation being strongly predicted by local recombination rates and the density of functional elements. This implies that the interaction between recurrent selection and intrinsic genomic features has dramatically sculpted the genomic landscape over long periods of time. In addition, our findings provide evidence that, apart from background selection, recent positive selection and long‐term balancing selection have also been crucial components in shaping patterns of genome‐wide variation during the speciation process.     

Competition and facilitation between tree species change with stand development

Processes governing tree interspecific interactions, such as facilitation and competition, may vary in strength over time. This study tried to unveil them by performing dendrometrical analyses on black spruce Picea mariana, trembling aspen Populus tremuloides and jack pine Pinus banksiana trees from pure and mixed mature boreal forest stands in the Clay Belt of northwestern Quebec and on the tills of northwestern Ontario. We cored 1430 trees and cut 120 for stem analysis across all stand composition types, tree species and study regions. Aspen annual growth rate was initially higher when mixed with conifers, but then progressively decreased over time compared to pure aspen stands, while jack pine growth rate did not differ with black spruce presence throughout all stages of stand development. When mixed with aspen, black spruce showed a contrary response to aspen, i.e. an initial loss in growth but a positive gain later. On the richer clay soil of the Quebec Clay Belt region, however, both aspen and spruce responses in mixed stands reversed between 37 and 54 years. Overall, our results demonstrate that interspecific interactions were present and tended to change with stand development and among species. Our results also suggest that the nature of interspecific interactions may differ with soil nutrient availability.     

Wood-decay type and fungal guild dominance across a North American log transplant experiment

We incubated 196 large-diameter aspen (Populus tremuloides), birch (Betula papyrifera), and pine (Pinus taeda) logs on the FACE Wood Decomposition Experiment encompassing eight climatically-distinct forest sites in the United States. We sampled dead wood from these large-diameter logs after 2 to 6 y of decomposition and determined wood rot type as a continuous variable using the lignin loss/density loss ratio (L/D) and assessed wood-rotting fungal guilds using high-throughput amplicon sequencing (HTAS) of the ITS-2 marker. We found L/D values in line with a white rot dominance in all three tree species, with pine having lower L/D values than aspen and birch. Based on HTAS data, white rot fungi were the most abundant and diverse wood-rotting fungal guild, and soft rot fungi were more abundant and diverse than brown rot fungi in logs with low L/D values. For aspen and birch logs, decay type was related to the wood density at sampling. For the pine logs, decay type was associated with the balance between white and brown/soft rot fungi abundance and OTU richness. Our results demonstrate that decay type is governed by biotic and abiotic factors, which vary by tree species.     

Imaging spectroscopy links aspen genotype with below-ground processes at landscape scales

Fine-scale biodiversity is increasingly recognized as important to ecosystem-level processes. Remote sensing technologies have great potential to estimate both biodiversity and ecosystem function over large spatial scales. Here, we demonstrate the capacity of imaging spectroscopy to discriminate among genotypes of Populus tremuloides (trembling aspen), one of the most genetically diverse and widespread forest species in North America. We combine imaging spectroscopy (AVIRIS) data with genetic, phytochemical, microbial and biogeochemical data to determine how intraspecific plant genetic variation influences below-ground processes at landscape scales. We demonstrate that both canopy chemistry and below-ground processes vary over large spatial scales (continental) according to aspen genotype. Imaging spectrometer data distinguish aspen genotypes through variation in canopy spectral signature. In addition, foliar spectral variation correlates well with variation in canopy chemistry, especially condensed tannins. Variation in aspen canopy chemistry, in turn, is correlated with variation in below-ground processes. Variation in spectra also correlates well with variation in soil traits. These findings indicate that forest tree species can create spatial mosaics of ecosystem functioning across large spatial scales and that these patterns can be quantified via remote sensing techniques. Moreover, they demonstrate the utility of using optical properties as proxies for fine-scale measurements of biodiversity over large spatial scales.     

Genetic linkage mapping in aspen (Populus tremula L. and Populus tremuloides Michx.)

A large number of simple sequence repeat (SSR) marker-containing genetic maps are available for several Populus species. For aspen however, no SSR-containing map has been published so far. In this study, genetic linkage mapping was carried out with an interspecific mapping pedigree of 61 full-sib hybrids of European × quaking aspen (Populus tremula L. × Populus tremuloides Michx.), using the two-way pseudo-testcross strategy. Amplified fragment-length polymorphism (AFLP) and SSR markers were used for mapping, resulting in the first SSR-containing genetic linkage maps for aspen. The maps allow comparisons with a Populus consensus map and other published genetic maps of the genus Populus. The maps showed good collinearity to each other and to the Populus consensus map and provide a direct link to the Populus trichocarpa genomic sequence. Sex as a morphological trait was assessed in the mapping population and mapped on a non-terminal position of linkage group XIX on the male P. tremuloides map.     

Influence of Aspen on Forest Floor Properties in Black Spruce-dominated Stands

In the absence of fire in black spruce-feathermoss stands, a thick forest floor layer dominated by bryophytes and sphagnum accumulates. This layer is associated with wet, cool and nutrient-poor soil conditions conducive to the paludification process and pushing the ecosystem towards an unproductive open black spruce forest. The presence of Populus tremuloides in theses stands may halt this process because this species has a high nutrient cycling rate and a litter that represses moss cover. The main hypothesis of this study is that, despite similar abiotic conditions (slope and drainage), the presence of Populus tremuloides in a stand dominated by Picea mariana affects surface soil nutrient availability, total N, pH as well as the decomposition process. The abundance of Populus tremuloides trees was associated with higher exchangeable cations, cationic exchangeable capacity and pH of the forest floor layer on all sites. A decrease in organic matter thickness with increasing aspen presence was also found on all sites, suggesting that this species affects the decomposition process by the quality of its litter as well as by a general improvement of soil physical and chemical properties. The decomposition rate of a standard substrate as well as in vitro potential net nitrogen mineralization were positively related to Populus tremuloides on only one of the three sites, and non-significant on the other sites. Strong immobilization of added nitrogen during incubation was observed on all sites and was not related to aspen, which suggested that in these stands, the soil microbial community is uniformly and strongly nitrogen limited. The zone of influence of Populus tremuloides was evaluated in areas around the soil sampling plot ranging from 3 to 7 m. The results revealed that this zone varies with soil properties. The results suggest that the presence of Populus tremuloides accelerate nutrient cycling, which could affect stand productivity to some extent.     

Effects of genotype and nutrient availability on phytochemistry of trembling aspen (Populus tremuloides Michx.) during leaf senescence

We documented temporal patterns in phytochemical composition of Populus tremuloides Michx. during leaf senescence, and the influence of genotype and soil nutrient availability on such patterns. Levels of foliar nitrogen, carbohydrates, phenolic glycosides and condensed tannins were quantified for four aspen genotypes grown in a common garden, with low and high levels of soil nutrients. Levels of all compounds tended to decline over time, although the magnitude of change was influenced by plant genotype and nutrient availability. Genetic variation in concentrations of phytochemicals was much greater for phenolic glycosides and tannins than for nitrogen and carbohydrates, and these phenolic signatures generally persisted through leaf abscission. Our results suggest that genotypic and nutrient effects on patterns of chemical change during senescence will likely influence the performance of late-season herbivores on aspen. Moreover, nutrient and especially genotypic variation in phytochemistry of abscised leaves is likely to affect litter decomposition rates.     

Genetic Identity of Populus tremuloides Litter Influences Decomposition and Nutrient Release in a Mixed Forest Stand

Recent research has shown that genetic variation can directly impact community and ecosystem level processes. Populus tremuloides (trembling aspen) is an extremely widespread and genetically diverse tree species important to many North American forest ecosystems. Using leaf litter from five genotypes grown in a common garden under two nutrient treatments, we tracked litter decomposition in a natural aspen stand for 1 year. Here we show that aspen leaf litter decomposes and releases carbon, nitrogen, and sulfur in relation to its genetic identity. In a secondary experiment, we show that the genetic diversity of aspen litter mixtures can influence decomposition, however weakly so. Overall, nutrient treatments influenced leaf litter decomposition the most, followed by genetic identity, and then by genetic diversity (if at all in some cases). In this widespread, genetically diverse, and dominant species, genetic variation within a single species is important to ecosystem functioning. The relatively weak effect of genetic diversity on the processes measured here does not preclude its importance to ecosystem functioning, but does suggest that genetic identity and composition are more important than genetic diversity per se.     

Responses of ectomycorrhizal Populus tremuloides and Betula papyrifera seedlings to salinity

Roots of trembling aspen (Populus tremuloides Michx.) and paper birch (Betula papyrifera Marsh.) seedlings were inoculated with Hebeloma crustuliniforme or Laccaria bicolor and treated with 25 mM NaCl for 6 weeks. Both tree species appeared to be relatively tolerant of the applied NaCl treatment and did not develop visible leaf symptoms that are characteristic of salt injury. Salt treatment reduced total dry weights in aspen and birch, but did not significantly affect transpiration rates and root hydraulic conductance. Salt-treated ectomycorrhizal aspen maintained higher root hydraulic conductance compared with non-mycorrhizal plants. Na and Cl concentrations increased in shoots and roots of mycorrhizal and non-mycorrhizal aspen and birch in response to NaCl treatment. Roots of NaCl-treated aspen inoculated with H. crustuliniforme had over twofold higher concentrations of Na compared with non-mycorrhizal NaCl-treated plants. Similarly to aspen, Na and Cl concentrations increased in roots and shoots of NaCl-treated birch seedlings. However, in birch, there were no significant differences in Na and Cl concentrations between mycorrhizal and non-mycorrhizal plants. The results suggest that salt exclusion by the ectomycorrhizal associations is host-specific or/and that the processes leading to salt exclusion are activated in ectomycorrhizal plants by a threshold salt level which may vary between plant species.     

Drought stress limits the geographic ranges of two tree species via different physiological mechanisms

Range shifts are among the most ubiquitous ecological responses to anthropogenic climate change and have large consequences for ecosystems. Unfortunately, the ecophysiological forces that constrain range boundaries are poorly understood, making it difficult to mechanistically project range shifts. To explore the physiological mechanisms by which drought stress controls dry range boundaries in trees, we quantified elevational variation in drought tolerance and in drought avoidance‐related functional traits of a widespread gymnosperm (ponderosa pine – Pinus ponderosa) and angiosperm (trembling aspen – Populus tremuloides) tree species in the southwestern USA. Specifically, we quantified tree‐to‐tree variation in growth, water stress (predawn and midday xylem tension), drought avoidance traits (branch conductivity, leaf/needle size, tree height, leaf area‐to‐sapwood area ratio), and drought tolerance traits (xylem resistance to embolism, hydraulic safety margin, wood density) at the range margins and range center of each species. Although water stress increased and growth declined strongly at lower range margins of both species, ponderosa pine and aspen showed contrasting patterns of clinal trait variation. Trembling aspen increased its drought tolerance at its dry range edge by growing stronger but more carbon dense branch and leaf tissues, implying an increased cost of growth at its range boundary. By contrast, ponderosa pine showed little elevational variation in drought‐related traits but avoided drought stress at low elevations by limiting transpiration through stomatal closure, such that its dry range boundary is associated with limited carbon assimilation even in average climatic conditions. Thus, the same climatic factor (drought) may drive range boundaries through different physiological mechanisms – a result that has important implications for process‐based modeling approaches to tree biogeography. Further, we show that comparing intraspecific patterns of trait variation across ranges, something rarely done in a range‐limit context, helps elucidate a mechanistic understanding of range constraints.     

High efficiency poplar transformation

With the completion of the poplar tree genome database, Populus species have become one of the most useful model systems for the study of woody plant biology. Populus tremuloides (quaking aspen) is the most wide-spread tree species in North America, and its rapid growth generates the most abundant wood-based biomass out of any other plant species. To study such beneficial traits, there is a need for easier and more efficient transformation procedures that will allow the study of large numbers of tree genes. We have developed transformation procedures that are suitable for high-throughput format transformations using either Agrobacterium tumefaciens to produce transformed trees or Agrobacterium rhizogenes to generate hairy roots. Our method uses Agrobacterium inoculated aspen seedling hypocotyls followed by direct thidiazuron (TDZ)-mediated shoot regeneration on selective media. Transformation was verified through β-glucuronidase (GUS) reporter gene expression in all tree tissues, PCR amplification of appropriate vector products from isolated genomic DNA, and northern hybridization of incorporated and expressed transgenes. The hairy root protocol follows the same inoculation procedures and was tested using GUS reporter gene integration and antibiotic selection. The benefit of these procedures is that they are simple and efficient, requiring no maintenance of starting materials and allowing fully formed transgenic trees (or hairy roots) to be generated in only 3–4 months, rather than the 6–12 months required by more traditional methods. Likewise, the fact that the protocols are amenable to high-throughput formats makes them better suited for large-scale functional genomics studies in poplars.     

Soil and tree chemistry reflected the cumulative impact of acid deposition in Pinus banksiana and Populus tremuloides stands in the Athabasca oil sands region in western Canada

The spatial variability of soil chemistry and Ca/Al ratios of soil solution and fine roots were investigated in jack pine (Pinus banksiana) and trembling aspen (Populus tremuloides, aspen) stands to assess the impact of chronic acid deposition on boreal forest ecosystems in the Athabasca oil sands region (AOSR) in Alberta, Canada. Available SO₄²⁻ (as the sum of soluble and adsorbed SO₄²⁻) accumulated in the soil near tree boles of both species, reflecting the influence of canopy intercepted SO₄²⁻. In jack pine stands, pH and soluble base cation concentrations decreased towards tree boles due to increased SO₄²⁻ leaching; the reverse was found in aspen stands due to deposition of base cations leached from the canopy. As a result, Ca/Al ratios in the soluble fraction in soils near jack pine boles were 5–20 times lower than that near aspen boles. The Ca/Al ratio did not reach the critical limits of 1.0 for soil solution (ranged from 1.0 to 4.1) or 0.5 for fine roots (0.7–7.9) in the studied watersheds. However, Alⁿ⁺ concentrations in the soil solution ranged from 0.2 to 4.1 mg L⁻¹ in NE7 and from 0.1 to 8.5 mg L⁻¹ in SM8 that can inhibit the growth of white spruce (Picea glauca) seedlings that commonly succeed aspen in upland sites in the AOSR. We suggest that the spatial variation caused by tree canopies/stems will affect forest regeneration and the effect of acid deposition on forest succession in the AOSR should be further studied.     

Diurnal and seasonal changes of leaf lamina hydraulic conductance in bur oak (Quercus macrocarpa) and trembling aspen (Populus tremuloides)

The aim of this study was to examine the diurnal and seasonal variations in the sensitivity of leaf lamina (K _lam) hydraulic conductance to irradiance in bur oak (Quercus macrocarpa Michx.) and trembling aspen (Populus tremuloides Michx.), which vary in their responses of K _lam to irradiance. K _lam was determined using the high-pressure method and the measurements were carried out in June, August and September. The irradiance response of K _lam in bur oak was present throughout the day and declined in senescing leaves. In trembling aspen, K _lam declined from morning to late afternoon and drastically decreased before the onset of leaf senescence, but it was not sensitive to irradiance. In both tree species, the capacity of the petioles to supply water to leaf lamina changed during the day in accordance with the ability of the leaf lamina to transport water. Petiole hydraulic conductivity (K _pet) declined during the season in bur oak leaves, while it tended to increase in trembling aspen leaves. There was no correlation between the K _lam values and air temperature or light intensity at the time of leaf collection. For trembling aspen, K _pet was negatively correlated with the air temperature suggesting sensitivity to drought. We conclude that the water transport properties of petioles and leaf lamina in the two studied tree species reflect their ecological adaptations. Trembling aspen leaves have high hydraulic conductivity and high stomatal conductance regardless of the irradiance level, consistent with the rapid growth and high demand for water. In contrast, the increased lamina hydraulic conductivity and stomatal conductance under high irradiance in bur oak trees reflect a water conservation strategy.     

Transcriptome Analysis of Poplar during Leaf Spot Infection with Sphaerulina spp.

Diseases of poplar caused by the native fungal pathogen Sphaerulina musiva and related species are of growing concern, particularly with the increasing interest in intensive poplar plantations to meet growing energy demands. Sphaerulina musiva is able to cause infection on leaves, resulting in defoliation and canker formation on stems. To gain a greater understanding of the different responses of poplar species to infection caused by the naturally co-evolved Sphaerulina species, RNA-seq was conducted on leaves of Populus deltoides, P. balsamifera and P. tremuloides infected with S. musiva, S. populicola and a new undescribed species, Ston1, respectively. The experiment was designed to contain the pathogen in a laboratory environment, while replicating disease development in commercial plantations. Following inoculation, trees were monitored for disease symptoms, pathogen growth and host responses. Genes involved in phenylpropanoid, terpenoid and flavonoid biosynthesis were generally upregulated in P. balsamifera and P. tremuloides, while cell wall modification appears to play an important role in the defense of P. deltoides. Poplar defensive genes were expressed early in P. balsamifera and P. tremuloides, but their expression was delayed in P. deltoides, which correlated with the rate of disease symptoms development. Also, severe infection in P. balsamifera led to leaf abscission. This data gives an insight into the large differences in timing and expression of genes between poplar species being attacked by their associated Sphaerulina pathogen.     

Photoperiodic growth control in perennial trees

Plants have to cope with changing seasons and adverse environmental conditions. Being sessile, plants have developed elaborate mechanisms for their survival that allow them to sense and adapt to the environment and reproduce successfully. A major adaptive trait for the survival of trees of temperate and boreal forests is the induction of growth cessation in anticipation of winters. In the last few years enormous progress has been made to elucidate the molecular mechanisms underlying SDs induced growth cessation in model perennial tree hybrid aspen (Populus tremula × P. tremuloides). In this review we discuss the molecular mechanism underlying photoperiodic control of growth cessation and adaptive responses.     

<Emphasis Type="Bold">Epigenetic variation in clonal stands of aspen</Emphasis>

Epigenetic mechanisms can affect ecologically important traits, even in the absence of genetic variation. Environmental factors can influence gene regulation through chemical modifications, such as DNA methylation, resulting in acclimation that can be transferred to subsequent cell generations both mitotically and meiotically. Clonal plants such as Populus tremuloides (aspen) show considerable promise as model species for the long-term in situ study of ecological epigenetics. The common replication of identical genotypes across heterogeneous environments permits within- and between-genotype comparisons while controlling for genetic makeup. With a long lifespan and limited natural selection resulting from sexual recombination, it is conceivable that epigenetic acclimation plays an important role in the long-term ecological success of aspen. This case study is the first in a series investigating the role of epigenetics in aspen ecology. We have established long-term permanent plots of aspen, identified (genotyped) clones and established the baseline epigenetic structure. Here we report the in situ epigenetic structure of two aspen stands. We find considerable epigenetic variation and significant differences within and among genotypes and sites, suggesting both genotype and environment influence the epigenotype.