Genetic variation in circadian regulation of nocturnal stomatal conductance enhances carbon assimilation and growth

Circadian resonance, whereby a plant's endogenous rhythms are tuned to match environmental cues, has been repeatedly shown to be adaptive, although the underlying mechanisms remain elusive. Concomitantly, the adaptive value of nocturnal transpiration in C₃ plants remains unknown because it occurs without carbon assimilation. These seemingly unrelated processes are interconnected because circadian regulation drives temporal patterns in nocturnal stomatal conductance, with maximum values occurring immediately before dawn for many species. We grew individuals of six Eucalyptus camaldulensis genotypes in naturally lit glasshouses and measured sunset, predawn and midday leaf gas exchange and whole‐plant biomass production. We tested whether sunrise anticipation by the circadian clock and subsequent increases in genotype predawn stomatal conductance led to rapid stomatal opening upon illumination, ultimately affecting genotype differences in carbon assimilation and growth. We observed faster stomatal responses to light inputs at sunrise in genotypes with higher predawn stomatal conductance. Moreover, early morning and midday stomatal conductance and carbon assimilation, leaf area and total plant biomass were all positively correlated with predawn stomatal conductance across genotypes. Our results lead to the novel hypothesis that genotypic variation in the circadian‐regulated capacity to anticipate sunrise could be an important factor underlying intraspecific variation in tree growth.     

Quantitative variation in water-use efficiency across water regimes and its relationship with circadian, vegetative, reproductive, and leaf gas-exchange traits

Drought limits light harvesting, resulting in lower plant growth and reproduction. One trait important for plant drought response is water-use efficiency (WUE). We investigated (1) how the joint genetic architecture of WUE, reproductive characters, and vegetative traits changed across drought and well-watered conditions, (2) whether traits with distinct developmental bases (e.g. leaf gas exchange versus reproduction) differed in the environmental sensitivity of their genetic architecture, and (3) whether quantitative variation in circadian period was related to drought response in Brassica rapa. Overall, WUE increased in drought, primarily because stomatal conductance, and thus water loss, declined more than carbon fixation. Genotypes with the highest WUE in drought expressed the lowest WUE in well-watered conditions, and had the largest vegetative and floral organs in both treatments. Thus, large changes in WUE enabled some genotypes to approach vegetative and reproductive trait optima across environments. The genetic architecture differed for gas-exchange and vegetative traits across drought and well-watered conditions, but not for floral traits. Correlations between circadian and leaf gas-exchange traits were significant but did not vary across treatments, indicating that circadian period affects physiological function regardless of water availability. These results suggest that WUE is important for drought tolerance in Brassica rapa and that artificial selection for increased WUE in drought will not result in maladaptive expression of other traits that are correlated with WUE.     

Independent variation in photosynthetic capacity and stomatal conductance leads to differences in intrinsic water use efficiency in 11 soybean genotypes before and during mild drought

Intrinsic water use efficiency (WUE(intr)), the ratio of photosynthesis to stomatal conductance to water, is often used as an index for crop water use in breeding projects. However, WUE(intr) conflates variation in these two processes, and thus may be less useful as a selection trait than knowledge of both components. The goal of the present study was to determine whether the contribution of photosynthetic capacity and stomatal conductance to WUE(intr) varied independently between soybean genotypes and whether this pattern was interactive with mild drought. Photosynthetic capacity was defined as the variation in WUE(intr) that would occur if genotypes of interest had the same stomatal conductance as a reference genotype and only differed in photosynthesis; similarly, the contribution of stomatal conductance to WUE(intr) was calculated assuming a constant photosynthetic capacity across genotypes. Genotypic differences in stomatal conductance had the greatest effect on WUE(intr) (26% variation when well watered), and was uncorrelated with the effect of photosynthetic capacity on WUE(intr). Thus, photosynthetic advantages of 8.3% were maintained under drought. The maximal rate of Rubisco carboxylation, generally the limiting photosynthetic process for soybeans, was correlated with photosynthetic capacity. As this trait was not interactive with leaf temperature, and photosynthetic capacity differences were maintained under mild drought, the observed patterns of photosynthetic advantage for particular genotypes are likely to be consistent across a range of environmental conditions. This suggests that it is possible to employ a selection strategy of breeding water-saving soybeans with high photosynthetic capacities to compensate for otherwise reduced photosynthesis in genotypes with lower stomatal conductance.     

Genetic variation in Arabidopsis thaliana for night-time leaf conductance

Night-time leaf conductance ( g _night) and transpiration may have several adaptive benefits related to plant water, nutrient and carbon relations. Little is known, however, about genetic variation in g _night and whether this variation correlates with other gas exchange traits related to water use and/or native habitat climate. We investigated g _night in 12 natural accessions and three near isogenic lines (NILs) of Arabidopsis thaliana . Genetic variation in g _night was found for the natural accessions, and g _night was negatively correlated with native habitat atmospheric vapour pressure deficit (VPD_air), suggesting lower g _night may be favoured by natural selection in drier habitats. However, there were also significant genetic correlations of g _night with daytime gas exchange traits expected to affect plant fitness [i.e. daytime leaf conductance, photosynthesis and intrinsic water-use efficiency (WUE_i)], indicating that selection on daytime gas exchange traits may result in indirect selection on g _night. The comparison of three NILs to their parental genotypes identified one quantitative trait locus (QTL) contributing to variation in g _night. Further characterization of genetic variation in g _night within and among populations and species, and of associations with other traits and native habitats will be needed to understand g _night as a putatively adaptive trait.     

Genetic variation in stomatal and biochemical limitations to photosynthesis in the annual plant, Polygonum arenastrum

 Terrestrial plant photosynthesis may be limited both by stomatal behavior and leaf biochemical capacity. While inferences have been made about the importance of stomatal and biochemical limitations to photosynthesis in a variety of species in a range of environments, genetic variation in these limitations has never been documented in wild plant populations. Genetic variation provides the raw material for adaptive evolution in rates of carbon assimilation. We examined genetic variation in gas exchange physiology and in stomatal and biochemical traits in 16 genetic lines of the annual plant, Polygonum arenastrum. The photosynthesis against leaf internal CO₂ (A−ci) response curve was measured on three greenhouse-grown individuals per line. We measured the photosynthetic rate (A) and stomatal conductance (g), and calculated the internal CO₂ concentration (ci) at ambient CO₂ levels. In addition, the following stomatal and biochemical characteristics were obtained from the A−ci curve on each individual: the degree of stomatal limitation to photosynthesis (L_s), the maximum ribulose 1,5-biphosphate carboxylase-oxygenase (Rubisco) activity (Vc_max) and electron transport capacity (J_max). All physiological traits were genetically variable, with broad sense heritabilities ranging from 0.66 for L_s to 0.94 for J_max. Strong positive genetic correlations were found between Vc_max and J_max, and between g and biochemical capacity. Path analyses revealed strong causal influences of stomatal conductance and leaf biochemistry on A and ci. Path analysis also indicated that L_s confounds both stomatal and biochemical effects, and is an appropriate measure of stomatal influences on photosynthesis, only when biochemical variation is accounted for. In total, our results indicate that differences among lines in photosynthesis and ci result from simultaneous changes in biochemical and stomatal characteristics and are consistent with theoretical predictions that there should be co-limitation of photosynthesis by ribulose-1,5-biphosphate (RuBP) utilization and regeneration, and by stomatal conductance and leaf biochemistry. Gas exchange characteristics of genetic lines in the present study were generally consistent with measurements of the same lines in a previous field study. Our new results indicate that the mechanisms underlying variation in gas exchange include variation in both stomatal conductance and biochemical capacity. In addition, A, g, and ci in the present study tended also to be positively correlated with carbon isotope discrimination (Δ), and negatively correlated with time to flowering, life span, and leaf size based on earlier work. The pattern of correlation between physiology and life span among genetic lines of P. arenastrum parallels interspecific patterns of character correlations. We suggest that the range of trait constellations among lines in P. arenastrum represents a continuum between stress avoidance (rapid development, high gas exchange metabolism) and stress tolerance (slow development, low gas exchange metabolism), and that genetic variation in these character combinations may be maintained by environmental variation in stress levels in the species’ ruderal habitat. Received: 28 March 1996 / Accepted: 13 August 1996     

Limiting factors for panicle photosynthesis at the anthesis and grain filling stages in rice (Oryza sativa L.)

Panicle photosynthesis is crucial for grain yield in cereal crops; however, the limiting factors for panicle photosynthesis are poorly understood, greatly impeding improvement in this trait. In the present study, pot experiments were conducted to investigate the limiting factors for panicle photosynthesis at the anthesis stage in seven rice genotypes and to examine the temporal variations in photosynthesis during the grain filling stage in the Liangyou 287 genotype. At the anthesis stage, leaf and panicle photosynthesis was positively correlated with stomatal conductance and maximum carboxylation rate, which were in turn associated with hydraulic conductance and nitrogen content, respectively. Panicle hydraulic conductance was positively correlated with the area of bundle sheaths in the panicle neck. During grain filling, leaf and panicle photosynthesis remained constant at the early stage but dramatically decreased from 8 to 9 days after anthesis. The trends of variations in panicle photosynthesis were consistent with those in stomatal conductance but not with those in maximum carboxylation rate. At first, the maximum carboxylation rate and respiration rate in the panicle increased, through elevated panicle nitrogen content, but then drastically decreased, as a result of dehydration. The present study systematically investigated the limiting factors for panicle photosynthesis, which are vital for improving photosynthesis and crop yield.     

Genetic variation of ecophysiological traits in two gynodioecious species of Schiedea (Caryophyllaceae)

Evolution of dimorphic breeding systems may involve changes in ecophysiological traits as well as floral morphology because of greater resource demands on females. Differences between related species suggest that ecophysiological traits should be heritable, and species with higher female frequencies should show greater sexual differentiation. We used modified partial diallel crossing designs to estimate narrow-sense heritabilities and genetic correlations of sex-specific ecophysiological and morphological traits in closely related gynodioecious Schiedea salicaria (13% females) and Schiedea adamantis (39% females). In S. salicaria, hermaphrodites and females differed in photosynthetic rate and specific leaf area (SLA). Narrow-sense heritabilities were significant for stomatal conductance, SLA and inflorescence number in hermaphrodites, and for SLA and inflorescence number in females. Schiedea adamantis had no sexual dimorphism in measured traits; stomatal conductance, stem number and inflorescence number were heritable in females, and stem number was heritable in hermaphrodites. In both species, significant genetic correlations of traits between sexes were rare, indicating that traits can evolve independently in response to sex-differential selection. Significant genetic correlations were detected between certain traits within sexes of both species. Low heritability of some ecophysiological traits may reflect low additive genetic variability or high phenotypic plasticity in these traits.     

Association genetics,geography and ecophysiology link stomatal patterning in Populus trichocarpa with carbon gain and disease resistance trade‐offs

Stomata are essential for diffusive entry of gases to support photosynthesis, but may also expose internal leaf tissues to pathogens. To uncover trade‐offs in range‐wide adaptation relating to stomata, we investigated the underlying genetics of stomatal traits and linked variability in these traits with geoclimate, ecophysiology, condensed foliar tannins and pathogen susceptibility in black cottonwood (Populus trichocarpa). Upper (adaxial) and lower (abaxial) leaf stomatal traits were measured from 454 accessions collected throughout much of the species range. We calculated broad‐sense heritability (H²) of stomatal traits and, using SNP data from a 34K Populus SNP array, performed a genome‐wide association studies (GWAS) to uncover genes underlying stomatal trait variation. H² values for stomatal traits were moderate (average H² = 0.33). GWAS identified genes associated primarily with adaxial stomata, including polarity genes (PHABULOSA), stomatal development genes (BRASSINOSTEROID‐INSENSITIVE 2) and disease/wound‐response genes (GLUTAMATE‐CYSTEINE LIGASE). Stomatal traits correlated with latitude, gas exchange, condensed tannins and leaf rust (Melampsora) infection. Latitudinal trends of greater adaxial stomata numbers and guard cell pore size corresponded with higher stomatal conductance (g_s) and photosynthesis (A_max), faster shoot elongation, lower foliar tannins and greater Melampsora susceptibility. This suggests an evolutionary trade‐off related to differing selection pressures across the species range. In northern environments, more adaxial stomata and larger pore sizes reflect selection for rapid carbon gain and growth. By contrast, southern genotypes have fewer adaxial stomata, smaller pore sizes and higher levels of condensed tannins, possibly linked to greater pressure from natural leaf pathogens, which are less significant in northern ecosystems.     

Natural variation,differentiation, and genetic trade‐offs of ecophysiological traits in response to water limitation in Brachypodium distachyon and its descendent allotetraploid B. hybridum (Poaceae)

Differences in tolerance to water stress may underlie ecological divergence of closely related ploidy lineages. However, the mechanistic basis of physiological variation governing ecogeographical cytotype segregation is not well understood. Here, using Brachypodium distachyon and its derived allotetraploid B. hybridum as model, we test the hypothesis that, for heteroploid annuals, ecological divergence of polyploids in drier environments is based on trait differentiation enabling drought escape. We demonstrate that under water limitation allotetraploids maintain higher photosynthesis and stomatal conductance and show earlier flowering than diploids, concordant with a drought‐escape strategy to cope with water stress. Increased heterozygosity and greater genetic variability and plasticity of polyploids could confer a superior adaptive capability. Consistent with these predictions, we document (1) greater standing within‐population genetic variation in water‐use efficiency (WUE) and flowering time in allotetraploids, and (2) the existence of (nonlinear) environmental clines in physiology across allotetraploid populations. Increased gas exchange and diminished WUE occurred at the driest end of the gradient, consistent with a drought‐escape strategy. Finally, we found that allotetraploids showed weaker genetic correlations than diploids congruous with the expectation of relaxed pleiotropic constraints in polyploids. Our results suggest evolutionary divergence of ecophysiological traits in each ploidy lineage.     

叶片结构的水力学特性对植物生理功能影响的研究进展

叶片是植物进行光合、呼吸、蒸腾作用的主要器官, 早期的研究主要集中于水分在叶片中的运输路径, 而对叶脉结构及其生态学意义研究甚少。近年来关于叶片叶脉结构、气孔结构的功能及叶片水力学特性的意义研究已经成为植物生理生态的研究热点。该文综述了叶脉的结构性状的指标(叶脉密度、直径、间距等), 叶片水力学结构特性对植物生长、水分运输、气体交换、光合作用等生理功能的影响, 及其与植物对干旱适应性之间的关系。叶脉结构是决定叶片生理功能的基础, 因此在未来的工作中应分析比较不同种类植物叶脉结构形态与导水、光合、呼吸、同化作用之间的关系, 建立植物茎干-枝-叶系统水力传导的机理性模型, 用以探索不同植物功能结构和高效用水生理生态学机制, 据此评估不同种类植物在未来气候情景下的地位。     

Circadian rhythms have insignificant effects on plant gas exchange under field conditions

Circadian rhythms in photosynthesis and stomatal conductance have been widely observed, but their possible adaptive significance is unknown. To determine whether such rhythms have a significant effect on the daily courses of carbon gain and/or water loss under field conditions, we obtained laboratory data on circadian rhythms in gas exchange of Saururus cernuus L., a wetland perennial. Using these data we modified a widely used mathematical model of photosynthesis and stomatal conductance by introducing the observed circadian‐rhythmic variation into the maximum rates of electron transport and carboxylation. We measured photosynthesis and stomatal conductance hourly on the same species growing naturally in the field and compared measured daily courses of photosynthesis and stomatal conductance with daily courses calculated using the model as originally formulated and also as modified to include circadian rhythms. The model fit the field data only slightly better when rhythms were included: the rhythms accounted for only about 1% of the observed daily carbon gain. Thus, these rhythms probably do not affect photosynthesis and stomatal conductance in the field.     

Leaf economics and hydraulic traits are decoupled in five species‐rich tropical‐subtropical forests

Leaf economics and hydraulic traits are critical to leaf photosynthesis, yet it is debated whether these two sets of traits vary in a fully coordinated manner or there is room for independent variation. Here, we tested the relationship between leaf economics traits, including leaf nitrogen concentration and leaf dry mass per area, and leaf hydraulic traits including stomatal density and vein density in five tropical‐subtropical forests. Surprisingly, these two suites of traits were statistically decoupled. This decoupling suggests that independent trait dimensions exist within a leaf, with leaf economics dimension corresponding to light capture and tissue longevity, and the hydraulic dimension to water‐use and leaf temperature maintenance. Clearly, leaf economics and hydraulic traits can vary independently, thus allowing for more possible plant trait combinations. Compared with a single trait dimension, multiple trait dimensions may better enable species adaptations to multifarious niche dimensions, promote diverse plant strategies and facilitate species coexistence.     

红树林植物叶片形态和解剖特征对叶肉导度、叶片导水率的影响

叶肉导度和叶片导水率是影响光合作用的两个重要过程,叶肉导度通过影响从气孔下腔到Rubisco酶位点的二氧化碳浓度梯度直接影响光合作用,而叶片导水率则通过影响水分供应或气孔行为来影响光合作用,然而对这两个生理过程之间的协同性研究较少。本研究选择9种红树林植物为研究对象,探讨盐生环境下植物叶肉导度和叶片导水率的协同性及其与叶片解剖结构特征之间的相关性。结果表明,9种红树林植物叶片导水率(0.78~5.83 mmol·m~(-2)·s~(-1)·MPa-1)、叶肉导度(0.06~0.36 mol·m~(-2)·s~(-1))、最大光合速率(7.23~23.71μmol·m~(-2)·s~(-1))等特征的差别较大;叶肉导度与最大光合速率呈显著正相关,而与比叶重无显著相关性,其原因是由于比叶重与叶片厚度、叶片密度不存在相关性;叶脉密度与气孔密度呈较强的相关性,说明红树林植物叶片水分运输与散失相关的叶片结构之间存在协同关系;叶片导水率不受叶脉密度影响,并且与叶肉导度、最大光合速率也不存在相关性,这很可能与红树林植物叶片的肉质化、有发达的储水组织有关,体现了红树林植物叶片结构和功能的特殊性。     

Model-assisted ideotyping reveals trait syndromes to adapt viticulture to a drier climate

Climate change is challenging the resilience of grapevine (Vitis), one of the most important crops worldwide. Adapting viticulture to a hotter and drier future will require a multifaceted approach including the breeding of more drought-tolerant genotypes. In this study, we focused on plant hydraulics as a multi-trait system that allows the plant to maintain hydraulic integrity and gas exchange rates longer under drought. We quantified a broad range of drought-related traits within and across Vitis species, created in silico libraries of trait combinations, and then identified drought tolerant trait syndromes. By modeling the maintenance of hydraulic integrity of current cultivars and the drought tolerant trait syndromes, we identified elite ideotypes that increased the amount of time they could experience drought without leaf hydraulic failure. Generally, elites exhibited a trait syndrome with lower stomatal conductance, earlier stomatal closure, and a larger hydraulic safety margin. We demonstrated that, when compared with current cultivars, elite ideotypes have the potential to decrease the risk of hydraulic failure across wine regions under future climate scenarios. This study reveals the syndrome of traits that can be leveraged to protect grapevine from experiencing hydraulic failure under drought and increase drought tolerance.

This article quantifies the diversity of hydraulic traits across grapevine genotypes and reveals the trait combinations that can help increase grapevine drought tolerance.     

On the persistence of memory: Soft clocks and terrestrial biosphere-atmosphere interactions

The circadian clock is considered a central “orchestrator” of gene expression and metabolism. Concomitantly, the circadian clock is considered of negligible influence in the field and beyond leaf levels, where direct physiological responses to environmental cues are considered the main drivers of diel fluctuations. I propose to bridge the gap across scales by examining current evidence on whether circadian rhythmicity in gas exchange is relevant for field settings and at the ecosystem scale. Nocturnal stomatal conductance and water fluxes appear to be influenced by a “hard” clock that may override the direct physiological responses to the environment. Tests on potential clock controls over photosynthetic carbon assimilation and daytime transpiration are scant yet, if present, could have a large impact on our current understanding and modeling of the exchanges of carbon dioxide and water between terrestrial ecosystems and the atmosphere.     

Stomatal kinetics and photosynthetic gas exchange along a continuum of isohydric to anisohydric regulation of plant water status

Species' differences in the stringency of stomatal control of plant water potential represent a continuum of isohydric to anisohydric behaviours. However, little is known about how quasi‐steady‐state stomatal regulation of water potential may relate to dynamic behaviour of stomata and photosynthetic gas exchange in species operating at different positions along this continuum. Here, we evaluated kinetics of light‐induced stomatal opening, activation of photosynthesis and features of quasi‐steady‐state photosynthetic gas exchange in 10 woody species selected to represent different degrees of anisohydry. Based on a previously developed proxy for the degree of anisohydry, species' leaf water potentials at turgor loss, we found consistent trends in photosynthetic gas exchange traits across a spectrum of isohydry to anisohydry. More anisohydric species had faster kinetics of stomatal opening and activation of photosynthesis, and these kinetics were closely coordinated within species. Quasi‐steady‐state stomatal conductance and measures of photosynthetic capacity and performance were also greater in more anisohydric species. Intrinsic water‐use efficiency estimated from leaf gas exchange and stable carbon isotope ratios was lowest in the most anisohydric species. In comparisons between gas exchange traits, species rankings were highly consistent, leading to species‐independent scaling relationships over the range of isohydry to anisohydry observed.     

Functional and evolutionary correlations of steep leaf angles in the mexical shrubland

In the evergreen shrubland vegetation of Mexico (mexical), most of the species are sclerophyllous woody plants with steep leaf angles. This architectural pattern has been interpreted as a strategy to cope with water shortages and high radiation. However, the current association between evergreenness and steep leaf angles across mexical plant species could be the result of an adaptive association achieved through correlated evolutionary change between both traits or, alternatively, may be the result of common evolutionary ancestry. In this study, we quantified leaf angle in 28 dominant species under a phylogenetic framework and evaluated the functional implications of the observed range of leaf angles in terms of leaf temperature, water potentials and transpiration by combining manipulative experiments restraining leaves horizontally with microclimatic and stomatal conductance measurements in selected species and energy balance calculations. Horizontally restrained leaves exhibited reduced water potentials and stomatal conductances, and significantly increased temperatures and transpiration rates. Steeply inclined leaves operated near air temperatures and could sustain relatively high stomatal conductances during the dry season since they were associated with low transpiration rates. Phylogenetic analyses showed that steep leaf angles evolved in a correlated fashion in evergreen species. The functional consequences of leaf angle together with the phylogenetic analysis indicate the adaptive nature of this trait which allows the evergreen species to cope with arid conditions and therefore to persist within the mexical community.     

Changes in stomatal conductance along grass blades reflect changes in leaf structure

Identifying the consequences of grass blade morphology (long, narrow leaves) on the heterogeneity of gas exchange is fundamental to an understanding of the physiology of this growth form. We examined acropetal changes in anatomy, hydraulic conductivity and rates of gas exchange in five grass species (including C(3) and C(4) functional types). Both stomatal conductance and photosynthesis increased along all grass blades despite constant light availability. Hydraulic efficiency within the xylem remained constant along the leaf, but structural changes outside the xylem changed in concert with stomatal conductance. Stomatal density and stomatal pore index remained constant along grass blades but interveinal distance decreased acropetally resulting in a decreased path length for water movement from vascular bundle to stomate. The increase in stomatal conductance was correlated with the decreased path length through the leaf mesophyll. A strong correlation between the distance from vascular bundles to stomatal pores and stomatal conductance has been identified across species; our results suggest this relationship also exists within individual leaves.     

Leaf traits and performance vary with plant age and water availability in Artemisia californica

Background and AimsLeaf functional traits are strongly tied to growth strategies and ecological processes across species, but few efforts have linked intraspecific trait variation to performance across ontogenetic and environmental gradients. Plants are believed to shift towards more resource-conservative traits in stressful environments and as they age. However, uncertainty as to how intraspecific trait variation aligns with plant age and performance in the context of environmental variation may limit our ability to use traits to infer ecological processes at larger scales.MethodsWe measured leaf physiological and morphological traits, canopy volume and flowering effort for Artemisia californica (California sagebrush), a dominant shrub species in the coastal sage scrub community, under conditions of 50, 100 and 150 % ambient precipitation for 3 years.Key ResultsPlant age was a stronger driver of variation in traits and performance than water availability. Older plants demonstrated trait values consistent with a more conservative resource-use strategy, and trait values were less sensitive to drought. Several trait correlations were consistent across years and treatments; for example, plants with high photosynthetic rates tended to have high stomatal conductance, leaf nitrogen concentration and light-use efficiency. However, the trade-off between leaf construction and leaf nitrogen evident in older plants was absent for first-year plants. While few traits correlated with plant growth and flowering effort, we observed a positive correlation between leaf mass per area and performance in some groups of older plants.ConclusionsOverall, our results suggest that trait sensitivity to the environment is most visible during earlier stages of development, after which intraspecific trait variation and relationships may stabilize. While plant age plays a major role in intraspecific trait variation and sensitivity (and thus trait-based inferences), the direct influence of environment on growth and fecundity is just as critical to predicting plant performance in a changing environment.