José Alberto Ramírez-Valiente

Phenotypic plasticity and local adaptation in leaf ecophysiological traits of 13 contrasting cork oak populations under different water availabilities

Plants distributed across a wide range of environmental conditions are submitted to differential selective pressures. Long-term selection can lead to the development of adaptations to the local environment, generating ecotypic differentiation. Additionally, plant species can cope with this environmental variability by phenotypic plasticity. In this study, we examine the importance of both processes in coping with environmental heterogeneity in the Mediterranean sclerophyllous cork oak Quercus suber. For this purpose, we measured growth and key functional traits at the leaf level in 9-year-old plants across 2 years of contrasting precipitation (2005 and 2006) in a common garden. Plants were grown from acorns originated from 13 populations spanning a wide range of climates along the distribution range of the species. The traits measured were: leaf size (LS), specific leaf area (SLA), carbon isotope discrimination (Delta(13)C) and leaf nitrogen content per unit mass (N(mass)). Inter-population differences in LS, SLA and Delta(13)C were found. These differences were associated with rainfall and temperature at the sites of origin, suggesting local adaptation in response to diverging climates. Additionally, SLA and LS exhibited positive responses to the increase in annual rainfall. Year effect explained 28% of the total phenotypic variance in LS and 2.7% in SLA. There was a significant genotype x environment interaction for shoot growth and a phenotypic correlation between the difference in shoot growth among years and the annual mean temperature at origin. This suggests that populations originating from warm sites can benefit more from wet conditions than populations from cool sites. Finally, we investigated the relationships between functional traits and aboveground growth by several regression models. Our results showed that plants with lower SLA presented larger aboveground growth in a dry year and plants with larger leaf sizes displayed larger growth rates in both years. Overall, the study supports the adaptive value of SLA and LS for cork oak under a Mediterranean climate and their potentially important role for dealing with varying temperature and rainfall regimes through both local adaptation and phenotypic plasticity.

Factors affecting cork oak growth under dry conditions: local adaptation and contrasting additive genetic variance within populations

Increased drought severity is expected in the Mediterranean Basin over the twenty-first century, but our understanding of the potential of most forest tree species to cope with it remains uncertain. In this study, (1) we examined the potential effects of long-term selection and the capacity to respond to future changes in selective pressures in three populations of cork oak (Quercus suber L.). For this purpose, we evaluated the response to dry conditions of 45 open-pollinated trees originating from populations in Morocco, Portugal, and Spain. Growth, leaf size, specific leaf area (SLA), carbon isotope discrimination (Δ13C), leaf nitrogen content (Nmass), and total chlorophyll content (Chlmass) were measured in 9-year-old plants. (2) We also investigated the relationships between functional traits and aboveground growth by regression models. Plants presenting larger and more sclerophyllous leaves (low SLA and high leaf thickness) exhibited higher growths, with results suggesting that these traits are subjected to divergent selection in this species. Heritability estimates were moderately high for Δ13C (0.43 ± 0.25–0.83 ± 0.31) and stem diameter (0.40 ± 0.15–0.71 ± 0.28) for the tree populations. For the rest of the traits (except for annual growth), heritability values varied among populations, particularly for height, leaf size, leaf thickness, and Nmass. Our results suggest that natural selection has led to local adaptations and has also affected the genetic variance intrapopulation in these cork oak populations, although studies with a higher number of populations should be carried out across different years. Additionally, the absence of significant genetic correlations and the fact that correlated traits did not undergo opposing selection provided little evidence for constraints on evolution caused by genetic correlations.

Climatic origins predict variation in photoprotective leaf pigments in response to drought and low temperatures in live oaks (Quercus series Virentes)

Climate is a major selective force in nature. Exploring patterns of inter- and intraspecific genetic variation in functional traits may explain how species have evolved and may continue evolving under future climate change. Photoprotective pigments play an important role in short-term responses to climate stress in plants but knowledge of their long-term role in adaptive processes is lacking. In this study, our goal was to determine how photoprotective mechanisms, morphological traits and their plasticity have evolved in live oaks (Quercus series Virentes) in response to different climatic conditions. For this purpose, seedlings originating from 11 populations from four live oak species (Quercus virginiana, Q. geminata, Q. fusiformis and Q. oleoides) were grown under contrasting common environmental conditions of temperature (tropical vs temperate) and water availability (droughted vs well-watered). Xanthophyll cycle pigments, anthocyanin accumulation, chlorophyll fluorescence parameters and leaf anatomical traits were measured. Seedlings originating from more mesic source populations of Q. oleoides and Q. fusiformis increased the xanthophyll de-epoxidation state under water-limiting conditions and showed higher phenotypic plasticity for this trait, suggesting adaptation to local climate. Likewise, seedlings originating from warmer climates had higher anthocyanin concentration in leaves under cold winter conditions but not higher de-epoxidation state. Overall, our findings suggest that (i) climate has been a key factor in shaping species and population differences in stress tolerance for live oaks, (ii) anthocyanins are used under cold stress in species with limited freezing tolerance and (iii) xanthophyll cycle pigments are used when photoprotection under drought conditions is needed.

Evolutionary trade-offs between drought resistance mechanisms across a precipitation gradient in a seasonally dry tropical oak (Quercus oleoides)

In seasonally dry tropical forest regions, drought avoidance during the dry season coupled with high assimilation rates in the wet season is hypothesized to be an advantageous strategy for forest trees in regions with severe and long dry seasons. In contrast, where dry seasons are milder, drought tolerance coupled with a conservative resource-use strategy is expected to maximize carbon assimilation throughout the year. Tests of this hypothesis, particularly at the intraspecific level, have been seldom conducted. In this study, we tested the extent to which drought resistance mechanisms and rates of carbon assimilation have evolved under climates with varying dry season length and severity within Quercus oleoidesCham. and Schlect., a tropical dry forest species that is widely distributed in Central America. For this purpose, we conducted a greenhouse experiment where seedlings originating from five populations that vary in rainfall patterns were grown under different watering treatments. Our results revealed that populations from xeric climates with more severe dry seasons exhibited large mesophyllous leaves (with high specific leaf area, SLA), and leaf abscission in response to drought, consistent with a drought-avoidance strategy. In contrast, populations from more mesic climates with less severe dry seasons had small and thick sclerophyllous leaves with low SLA and reduced water potential at the turgor loss point (πtlp), consistent with a drought-tolerance strategy. Mesic populations also showed high plasticity in πtlp in response to water availability, indicating that osmotic adjustment to drought is an important component of this strategy. However, populations with mesophyllous leaves did not have higher maximum carbon assimilation rates under well-watered conditions. Furthermore, SLA was negatively associated with mass-based photosynthetic rates, contrary to expectations of the leaf economics spectrum, indicating that drought-resistance strategies are not necessarily tightly coupled with resource-use strategies. Overall, our study demonstrates the importance of considering intraspecific variation in analyses of the vulnerability of tropical trees to climate change.

Understanding the importance of intrapopulation functional variability and phenotypic plasticity in Quercus suber

Climate is one of the main abiotic factors influencing natural selection patterns. Year-to-year variation in climate is postulated to elicit temporal shifts in the direction and form of selection. Here, we examine the importance of trait means and plasticities for fitness under interannual variation in rainfall and assess the shifts in selection in cork oak. We performed selection analyses using the progeny of 45 mother trees established in a common garden experiment across two consecutive years that differed in rainfall. Growth and seven functional traits (specific leaf area, leaf size, leaf shape traits, 13-carbon isotope discrimination, and leaf nitrogen) related to drought tolerance were measured. Selection analyses showed fitness benefits of reduced specific leaf area (SLA) in a dry year and increased leaf size in a mesic year, indicating that they are key traits for this evergreen oak to cope with different water availabilities. SLA and leaf size were also particularly plastic traits, but the adaptive significance of plasticity could not be confirmed. The absence of correlation between growth across years using family means and the absence of correlations between SLA and leaf size suggested that fluctuating selection over time favored different maternal families under different annual weather conditions, which could promote functional diversity within populations in this long-lived species.

Population-Level Differentiation in Growth Rates and Leaf Traits in Seedlings of the Neotropical Live Oak Quercus oleoides Grown under Natural and Manipulated Precipitation Regimes

Widely distributed species are normally subjected to spatial heterogeneity in environmental conditions. In sessile organisms like plants, adaptive evolution and phenotypic plasticity of key functional traits are the main mechanisms through which species can respond to environmental heterogeneity and climate change. While extended research has been carried out in temperate species in this regard, there is still limited knowledge as to how species from seasonally-dry tropical climates respond to spatial and temporal variation in environmental conditions. In fact, studies of intraspecific genetically-based differences in functional traits are still largely unknown and studies in these ecosystems have largely focused on in situ comparisons where environmental and genetic effects cannot be differentiated. In this study, we tested for ecotypic differentiation and phenotypic plasticity in leaf economics spectrum (LES) traits, water use efficiency and growth rates under natural and manipulated precipitation regimes in a common garden experiment where seedlings of eight populations of the neotropical live oak Quercus oleoides were established. We also examined the extent to which intraspecific trait variation was associated with plant performance under different water availability. Similar to interspecific patterns among seasonally-dry tropical tree species, live oak populations with long and severe dry seasons had higher leaf nitrogen content and growth rates than mesic populations, which is consistent with a “fast” resource-acquisition strategy aimed to maximize carbon uptake during the wet season. Specific leaf area (SLA) was the best predictor of plant performance, but contrary to expectations, it was negatively associated with relative and absolute growth rates. This observation was partially explained by the negative association between SLA and area-based photosynthetic rates, which is contrary to LES expectations but similar to other recent intraspecific studies on evergreen oaks. Overall, our study shows strong intraspecific differences in functional traits in a tropical oak, Quercus oleoides, and suggests that precipitation regime has played an important role in driving adaptive divergence in this widespread species.

Adaptive consequences of human-mediated introgression for indigenous tree species: the case of a relict Pinus pinaster population

Human-induced gene movement via afforestation and restoration programs is a widespread phenomenon throughout the world. However, its effects on the genetic composition of native populations have received relatively little attention, particularly in forest trees. Here, we examine to what extent gene flow from allochthonous plantations of Pinus pinaster Aiton impacts offspring performance in a neighboring relict natural population and discuss the potential consequences for the long-term genetic composition of the latter. Specifically, we conducted a greenhouse experiment involving two contrasting watering treatments to test for differences in a set of functional traits and mortality rates between P. pinaster progenies from three different parental origins: (i) local native parents, (ii) exotic parents and (iii) intercrosses between local mothers and exotic fathers (intraspecific hybrids). Our results showed differences among crosses in cumulative mortality over time: seedlings of exotic parents exhibited the lowest mortality rates and seedlings of local origin the highest, while intraspecific hybrids exhibited an intermediate response. Linear regressions showed that seedlings with higher water-use efficiency (WUE, δ(13)C) were more likely to survive under drought stress, consistent with previous findings suggesting that WUE has an important role under dry conditions in this species. However, differences in mortality among crosses were only partially explained by WUE. Other non-measured traits and factors such as inbreeding depression in the relict population are more likely to explain the lower performance of native progenies. Overall, our results indicated that intraspecific hybrids and exotic individuals are more likely to survive under stressful conditions than local native individuals, at least during the first year of development. Since summer drought is the most important demographic and selective filter affecting tree establishment in Mediterranean ecosystems, a potential early selective advantage of exotic and hybrid genotypes would enhance initial steps of introgression of non-native genes into the study relict population of P. pinaster.

Natural selection and neutral evolutionary processes contribute to genetic divergence in leaf traits across a precipitation gradient in the tropical oak Quercus oleoides

The impacts of drought are expanding worldwide as a consequence of climate change. However, there is still little knowledge of how species respond to long‐term selection in seasonally dry ecosystems. In this study, we used QST‐FST comparisons to investigate (i) the role of natural selection on population genetic differentiation for a set of functional traits related to drought resistance in the seasonally dry tropical oak Quercus oleoides and (ii) the influence of water availability at the site of population origin and in experimental treatments on patterns of trait divergence. We conducted a thorough phenotypic characterization of 1912 seedlings from ten populations growing in field and greenhouse common gardens under replicated watering treatments. We also genotyped 218 individuals from the same set of populations using eleven nuclear microsatellites. QST distributions for leaf lamina area, specific leaf area, leaf thickness and stomatal pore index were higher than FST distribution. Results were consistent across growth environments. Genetic differentiation among populations for these functional traits was associated with the index of moisture at the origin of the populations. Together, our results suggest that drought is an important selective agent for Q. oleoides and that differences in length and severity of the dry season have driven the evolution of genetic differences in functional traits.

Evolutionary potential varies across populations and traits in the neotropical oak Quercus oleoides

Heritable variation in polygenic (quantitative) traits is critical for adaptive evolution and is especially important in this era of rapid climate change. In this study, we examined the levels of quantitative genetic variation of populations of the tropical tree Quercus oleoides Cham. and Schlect. for a suite of traits related to resource use and drought resistance. We tested whether quantitative genetic variation differed across traits, populations and watering treatments. We also tested potential evolutionary factors that might have shaped such a pattern: selection by climate and genetic drift. We measured 15 functional traits on 1322 1-year-old seedlings of 84 maternal half-sib families originating from five populations growing under two watering treatments in a greenhouse. We estimated the additive genetic variance, coefficient of additive genetic variation and narrow-sense heritability for each combination of traits, populations and treatments. In addition, we genotyped a total of 119 individuals (with at least 20 individuals per population) using nuclear microsatellites to estimate genetic diversity and population genetic structure. Our results showed that gas exchange traits and growth exhibited strikingly high quantitative genetic variation compared with traits related to leaf morphology, anatomy and photochemistry. Quantitative genetic variation differed between populations even at geographical scales as small as a few kilometers. Climate was associated with quantitative genetic variation, but only weakly. Genetic structure and diversity in neutral markers did not relate to coefficient of additive genetic variation. Our study demonstrates that quantitative genetic variation is not homogeneous across traits and populations of Q. oleoides. More importantly, our findings suggest that predictions about potential responses of species to climate change need to consider population-specific evolutionary characteristics.

Physiological Evidence from Common Garden Experiments for Local Adaptation and Adaptive Plasticity to Climate in American Live Oaks ( Quercus Section Virentes ): Implications for Conservation Under Global Change

Climate is known to be a critical factor controlling the broad-scale distribution of plants but often the physiological basis for species distribution limits is not well understood, nor is the extent to which populations within species are locally adapted to climate. Reciprocal transplant experiments designed to test for local adaptation are difficult to conduct and interpret in long-lived species, like oaks. Linking the physiological tolerances of species to their climatic distributions is an alternative approach to understanding adaptation to climate, and is important in predicting future distributions of species under changing climatic conditions. Here we synthesize a series of studies in a single lineage of American oaks that span the temperate tropical divide and encompass a range of precipitation and edaphic regimes, to determine (1) the physiological basis for adaptation to seasonal winter and seasonal drought and (2) the variation among populations that associated with climate variation and can be interpreted as local adaptation. We focus primarily on a series of common gardens that allow us to determine the genetically based differences in functional and physiological traits as well as the genetically based responses to contrasting temperature or precipitation regimes. We show that variation in freezing tolerance among closely related species is greater than variation among populations within species. Nevertheless, freezing tolerance varies predictably with climate of origin and is negatively associated with growth rate. In contrast, drought tolerance mechanisms vary more among populations within a single species, at least for the most widely distributed species, Quercus oleoides, than between species. Within this species, climate of origin predicts a suite of leaf physiological traits, and there is evidence for evolutionary trade-off between desiccation avoidance and desiccation resistance. Combined, these results show evidence for local adaptation to both freezing and drought stress within species, as well as adaptive differentiation between closely related species, despite phylogenetic conservatism in functional traits and highly similar physiognomy across the American live oak clade. The results inform conservation efforts aimed at preventing extinction of tree species in the face of global change.