Bridging the explanatory gaps: What can we learn from a biological agency perspective?

We begin this article by delineating the explanatory gaps left by prevailing gene-focused approaches in our understanding of phenotype determination, inheritance, and the origin of novel traits. We aim not to diminish the value of these approaches but to highlight where their implementation, despite best efforts, has encountered persistent limitations. We then discuss how each of these explanatory gaps can be addressed by expanding research foci to take into account biological agency-the capacity of living systems at various levels to participate in their own development, maintenance, and function by regulating their structures and activities in response to conditions they encounter. Here we aim to define formally what agency and agents are and-just as importantly-what they are not, emphasizing that agency is an empirical property connoting neither intention nor consciousness. Lastly, we discuss how incorporating agency helps to bridge explanatory gaps left by conventional approaches, highlight scientific fields in which implicit agency approaches are already proving valuable, and assess the opportunities and challenges of more systematically incorporating biological agency into research programs.

Niche construction and the environmental term of the price equation: How natural selection changes when organisms alter their environments.

Organisms construct their own environments and phenotypes through the adaptive processes of habitat choice, habitat construction, and phenotypic plasticity. We examine how these processes affect the dynamics of mean fitness change through the environmental change term of the Price Equation. This tends to be ignored in evolutionary theory, owing to the emphasis on the first term describing the effect of natural selection on mean fitness (the additive genetic variance for fitness of Fisher's Fundamental Theorem). Using population genetic models and the Price Equation, we show how adaptive niche constructing traits favorably alter the distribution of environments that organisms encounter and thereby increase population mean fitness. Because niche-constructing traits increase the frequency of higher-fitness environments, selection favors their evolution. Furthermore, their alteration of the actual or experienced environmental distribution creates selective feedback between niche constructing traits and other traits, especially those with genotype-by-environment interaction for fitness. By altering the distribution of experienced environments, niche constructing traits can increase the additive genetic variance for such traits. This effect accelerates the process of overall adaption to the niche-constructed environmental distribution and can contribute to the rapid refinement of alternative phenotypic adaptations to different environments. Our findings suggest that evolutionary biologists revisit and reevaluate the environmental term of the Price Equation: owing to adaptive niche construction, it contributes directly to positive change in mean fitness; its magnitude can be comparable to that of natural selection; and, when there is fitness G × E, it increases the additive genetic variance for fitness, the much-celebrated first term.

The relative impact of parental and current environment on plant transcriptomes depends on type of stress and genotype.

Through developmental plasticity, an individual organism integrates influences from its immediate environment with those due to the environment of its parents. While both effects on phenotypes are well documented, their relative impact has been little studied in natural systems, especially at the level of gene expression. We examined this issue in four genotypes of the annual plant Persicaria maculosa by varying two key resources-light and soil moisture-in both generations. Transcriptomic analyses showed that the relative effects of parent and offspring environment on gene expression (i.e. the number of differentially expressed transcripts, DETs) varied both for the two types of resource stress and among genotypes. For light, immediate environment induced more DETs than parental environment for all genotypes, although the precise proportion of parental versus immediate DETs varied among genotypes. By contrast, the relative effect of soil moisture varied dramatically among genotypes, from 8-fold more DETs due to parental than immediate conditions to 10-fold fewer. These findings provide evidence at the transcriptomic level that the relative impacts of parental and immediate environment on the developing organism may depend on the environmental factor and vary strongly among genotypes, providing potential for the interplay of these developmental influences to evolve.

DNA methylation mediates genetic variation for adaptive transgenerational plasticity.

Environmental stresses experienced by individual parents can influence offspring phenotypes in ways that enhance survival under similar conditions. Although such adaptive transgenerational plasticity is well documented, its transmission mechanisms are generally unknown. One possible mechanism is environmentally induced DNA methylation changes. We tested this hypothesis in the annual plant Polygonum persicaria, a species known to express adaptive transgenerational plasticity in response to parental drought stress. Replicate plants of 12 genetic lines (sampled from natural populations) were grown in dry versus moist soil. Their offspring were exposed to the demethylating agent zebularine or to control conditions during germination and then grown in dry soil. Under control germination conditions, the offspring of drought-stressed parents grew longer root systems and attained greater biomass compared with offspring of well-watered parents of the same genetic lines. Demethylation removed these adaptive developmental effects of parental drought, but did not significantly alter phenotypic expression in offspring of well-watered parents. The effect of demethylation on the expression of the parental drought effect varied among genetic lines. Differential seed provisioning did not contribute to the effect of parental drought on offspring phenotypes. These results demonstrate that DNA methylation can mediate adaptive, genotype-specific effects of parental stress on offspring phenotypes.

Evolutionary potential for increased invasiveness: High-Performance Polygonum cespitosum genotypes are competitively superior in full sun.

PREMISE OF STUDY: The presence of genetic variation for traits that contribute to ecological range expansion can provide the potential for introduced taxa to evolve greater invasiveness. Genotypes that contribute to the spread of introduced range populations must have the ability to maintain fitness under changing environmental stress and competitive intensity. Previously, we identified a subset of genotypes in populations of the invasive annual Polygonum cespitosum that express consistently high reproductive fitness in diverse (shaded, dry, and resource-rich) conditions. Here, we investigated whether these broadly adaptive (High-Performance) genotypes also show a competitive advantage over conspecifics in full sun and/or shade. METHODS: We grew a population-balanced sample of 13 High-Performance and 13 'Control' genotypes in intraspecific competitive arrays, comprising all four possible combinations of High-Performance vs. Control target plants and competitive backgrounds, in both full sun and shaded glasshouse environments. KEY RESULTS: In full sun, High-Performance genotypes (1) better maintained growth and reproductive output despite competition and (2) more strongly suppressed growth and reproduction of target plants. However, genotypes did not differ significantly in shade. CONCLUSIONS: Competitive superiority in open conditions may contribute to increasing predominance of these broadly adapted genotypes in introduced-range Polygonum cespitosum populations, and hence to the evolution of greater invasiveness. This study provides insight into the role of genotypic variation for ecological traits in the range expansion of a contemporary plant invader. It also highlights how such variation can be differently expressed in alternative environments (gene by environment interaction).

Limits to Future Adaptation in the Invasive Plant Polygonum cespitosum: Expression of Functional and Fitness Traits at Elevated CO2.

For organisms to adapt to future environments, they must both evolve appropriate functional responses and phenotypically express those responses under future climatic and CO2 conditions. We examined these 2 components of future adaptation in an invasive annual plant (Polygonum cespitosum) by performing a "resurrection" experiment under field conditions simulating a future environment. Resurrection experiments reveal recent evolution by comparing genotypes from natural populations sampled across a multigeneration interval. We collected genotypes from the same 3 North American populations in 1994 and 2005 and raised inbred lines from these collections under free air CO2 enrichment to examine functional and fitness traits expressed in hot, dry conditions at both ambient and elevated CO2 (N = 295 plants). The species has rapidly evolved in its introduced range to increase photosynthetic rate (collection year effect P ≤ 0.011) and delay senescence (P = 0.017) under full-sun, dry field conditions, but these adaptive changes were not expressed when the field environment included elevated CO2 (within-treatment year effect P ≥ 0.20 for both traits). Populations showed different levels of reproductive output and its genetic variance in these novel, stressful conditions. These findings illustrate constraints on evolutionary adaptation to predicted future conditions at both the species and population levels.

High-performance genotypes in an introduced plant: insights to future invasiveness.

Maintaining high reproductive output in diverse conditions has consistently been found to promote invasiveness in introduced taxa. Following on this key observation, studies have compared the performance across environments of invasive vs. native congeners, and of introduced vs. native populations within invasive species. Performance differences among genotypes within introduced species have received far less attention, although such genetic variation could be critical to invasive potential. If an introduced species contains genotypes that can maintain high fitness across contrasting environments, such broadly adaptive, high-performance genotypes could promote and shape the species' immediate spread across multiple habitats. Furthermore, their presence could lead to the evolution of greater aggressiveness in the species, as these high performers increase in frequency. We investigated the existence and distribution of high-performance genotypes in Polygonum cespitosum, a newly invasive Asian annual. We raised 416 genotypes, collected from 14 North American populations, under resource-rich conditions to identify potential high-performance genotypes (the top 5% in total reproductive output). We then compared their fitness, life history, and functional traits to a random group of the remaining genotypes in three contrasting environments to ask the following: (1) Do consistently high-performance genotypes (i.e., genotypes with high relative fitness in diverse conditions) exist within introduced-range populations? (2) If so, do these high-performance genotypes possess distinctive life history and/or functional traits? (3) Do these genotypes occur in all populations or in only a subset of populations? Genotypes initially identified as high-performance in favorable conditions also had higher reproductive output in resource-limited environments. Their fitness advantage compared with control genotypes varied in magnitude from one environment to another but was significant within all three test environments. High-performance genotypes shared a developmental syndrome characterized by rapid and high germination, fast seedling growth, early reproductive onset, and high reproductive allocation, but they did not differ in other functional traits. P. cespitosum includes a subset of genotypes with accelerated development and significantly greater fitness in both favorable and stressful conditions. The nonrandom distribution of these high-performance genotypes among populations in the species' introduced range highlights the importance of genotypic and population-level variation for invasion dynamics.

Post-introduction evolution of a rapid life-history strategy in a newly invasive plant.

A central question in invasion biology is whether adaptive trait evolution following species introduction promotes invasiveness. A growing number of common-garden experiments document phenotypic differences between native- and introduced-range plants, suggesting that adaptive evolution in the new range may indeed contribute to the success of invasive plants. However, these studies are often subject to methodological pitfalls, resulting in weak evidence for post-introduction adaptive trait evolution and leaving its role in the invasion process uncertain. In a common-garden glasshouse study, we compared the growth, life-history, and reproductive traits of 35 native- and introduced-range Polygonum cespitosum populations. We used complementary approaches including climate-matching, standardizing parental conditions, selection analysis, and testing for trait-environment relationships to determine whether traits that increase invasiveness adaptively evolved in the species' new range. We found that the majority of introduced-range populations exhibited a novel trait syndrome consisting of a fast-paced life history and concomitant sparse, reduced growth form. Selection analysis confirmed that this trait syndrome led to markedly higher fitness (propagule production) over a limited growing season that was characteristic of regions within the introduced range. Additionally, several growth and reproductive traits showed temperature-based clines consistent with adaptive evolution in the new range. Combined, these results indicate that, subsequent to its introduction to North America over 100 generations ago, P. cespitosum has evolved key traits that maximize propagule production. These changes may in part explain the species' recent transition to invasiveness, illustrating how post-introduction evolution may contribute to the invasion process.

Development and characterization of microsatellite markers for Polygonum cespitosum (Polygonaceae).

PREMISE OF THE STUDY: We isolated and characterized microsatellite markers in Polygonum cespitosum Blume, an herbaceous annual plant species introduced into North America from Asia that has recently become invasive. METHODS AND RESULTS: A total of 12 polymorphic and 3 monomorphic loci were screened in 1-2 individuals from each of 20 populations from the introduced and native range, for a total of 24 samples. The number of alleles per locus in the polymorphic loci ranged from 3 to 9, and expected heterozygosity ranged from 0.156 to 0.838. CONCLUSIONS: These new loci will provide tools for examining genetic relatedness among introduced and native populations of this and other related species.

Allopolyploid speciation in Persicaria (Polygonaceae): insights from a low-copy nuclear region.

Using a low-copy nuclear gene region (LEAFY second intron) we show multiple instances of allopolyploid speciation in Persicaria (Polygonaceae), which includes many important weeds. Fifteen species seem to be allopolyploids, which is higher than the number found in previous comparisons of chloroplast DNA and nuclear ribosomal internal transcribed spacer (nrITS) phylogenies. This underestimation of the extent of allopolyploidy is due in at least three cases to homogenization of nrITS toward the maternal lineage. One of the diploid species, P. lapathifolia, has been involved in at least six cases of allopolyploid speciation. Of the diploids, this species is the most widespread geographically and ecologically and also bears more numerous and conspicuous flowers, illustrating ecologic factors that may influence hybridization frequency. With a few exceptions, especially the narrowly endemic hexaploid, P. puritanorum, the allopolyploid species also are widespread, plastic, ecological generalists. Hybridization events fostered by human introductions may be fueling the production of new species that have the potential to become aggressive weeds.

Transgenerational effects of parent plant competition on offspring development in contrasting conditions.

Conditions during a parent's lifetime can induce phenotypic changes in offspring, providing a potentially important source of variation in natural populations. Yet, to date, biotic factors have seldom been tested as sources of transgenerational effects in plants. In a greenhouse experiment with the generalist annual Polygonum persicaria, we tested for effects of parental competition on offspring by growing isogenic parent plants either individually or in competitive arrays and comparing their seedling progeny in contrasting growth environments. Offspring of competing vs. non-competing parents showed significantly altered development, resulting in greater biomass and total leaf area, but only when growing in neighbor or simulated canopy shade, rather than sunny dry conditions. A follow-up experiment in which parent plants instead competed in dry soil found that offspring in dry soil had slightly reduced growth, both with and without competitors. In neither experiment were effects of parental competition explained by changes in seed provisioning, suggesting a more complex mode of regulatory inheritance. We hypothesize that parental competition in moist soil (i.e., primarily for light) confers specific developmental effects that are beneficial for light-limited offspring, while parental competition in dry soil (i.e., primarily for belowground resources) produces offspring of slightly lower overall quality. Together, these results indicate that competitive conditions during the parental generation can contribute significantly to offspring variation, but these transgenerational effects will depend on the abiotic resources available to both parents and progeny.

Developmental plasticity and human health.

Many plants and animals are capable of developing in a variety of ways, forming characteristics that are well adapted to the environments in which they are likely to live. In adverse circumstances, for example, small size and slow metabolism can facilitate survival, whereas larger size and more rapid metabolism have advantages for reproductive success when resources are more abundant. Often these characteristics are induced in early life or are even set by cues to which their parents or grandparents were exposed. Individuals developmentally adapted to one environment may, however, be at risk when exposed to another when they are older. The biological evidence may be relevant to the understanding of human development and susceptibility to disease. As the nutritional state of many human mothers has improved around the world, the characteristics of their offspring--such as body size and metabolism--have also changed. Responsiveness to their mothers' condition before birth may generally prepare individuals so that they are best suited to the environment forecast by cues available in early life. Paradoxically, however, rapid improvements in nutrition and other environmental conditions may have damaging effects on the health of those people whose parents and grandparents lived in impoverished conditions. A fuller understanding of patterns of human plasticity in response to early nutrition and other environmental factors will have implications for the administration of public health.

Understanding 'Non-genetic' Inheritance: Insights from Molecular-Evolutionary Crosstalk.

Understanding the evolutionary and ecological roles of 'non-genetic' inheritance (NGI) is daunting due to the complexity and diversity of epigenetic mechanisms. We draw on insights from molecular and evolutionary biology perspectives to identify three general features of 'non-genetic' inheritance systems: (i) they are functionally interdependent with, rather than separate from, DNA sequence; (ii) precise mechanisms vary phylogenetically and operationally; and (iii) epigenetic elements are probabilistic, interactive regulatory factors and not deterministic 'epialleles' with defined genomic locations and effects. We discuss each of these features and offer recommendations for future empirical and theoretical research that implements a unifying inherited gene regulation (IGR) approach to studies of 'non-genetic' inheritance.

Contrasting patterns of transgenerational plasticity in ecologically distinct congeners.

Stressful parental environments can influence offspring size and development either adaptively or maladaptively, yet little is known about species' differences in this complex aspect of phenotypic plasticity. We performed a reciprocal split-brood experiment to compare transgenerational plasticity in response to drought stress in two closely related annual plant species. We raised inbred replicate parent plants of eight genotypes per species in dry vs. moist soil to generate offspring of each genetic line that differed only in parental environment, then monitored seedling development in both dry and moist conditions. Individuals of the two species expressed contrasting patterns of transgenerational plasticity for traits important to seedling drought tolerance. In Polygonum persicaria, a weedy generalist found in moist, dry, and variably dry sites, drought-stressed plants produced offspring with longer and more rapidly extending root systems and greater biomass when growing in dry soil. In contrast, in P. hydropiper, a non-weedy congener restricted to moist habitats, the offspring of drought-stressed parents had reduced root system development and seedling biomass in dry soil. In P. persicaria, transgenerational and immediate adaptive plasticity combined to produce drought-adapted seedling phenotypes. These results make clear that characteristic patterns of transgenerational plasticity can contribute to ecological diversity among species.

Transgenerational effects of parental light environment on progeny competitive performance and lifetime fitness.

Plant and animal parents may respond to environmental conditions such as resource stress by altering traits of their offspring via heritable non-genetic effects. While such transgenerational plasticity can result in progeny phenotypes that are functionally pre-adapted to the inducing environment, it is unclear whether such parental effects measurably enhance the adult competitive success and lifetime reproductive output of progeny, and whether they may also adversely affect fitness if offspring encounter contrasting conditions. In glasshouse experiments with inbred genotypes of the annual plant Polygonum persicaria, we tested the effects of parental shade versus sun on (a) competitive performance of progeny in shade, and (b) lifetime reproductive fitness of progeny in three contrasting treatments. Shaded parents produced offspring with increased fitness in shade despite competition, as well as greater competitive impact on plant neighbours. Inherited effects of parental light conditions also significantly altered lifetime fitness: parental shade increased reproductive output for progeny in neighbour and understorey shade, but decreased fitness for progeny in sunny, dry conditions. Along with these substantial adaptive and maladaptive transgenerational effects, results show complex interactions between genotypes, parent environment and progeny conditions that underscore the role of environmental variability and change in shaping future adaptive potential. This article is part of the theme issue 'The role of plasticity in phenotypic adaptation to rapid environmental change'.

Context-Dependent Developmental Effects of Parental Shade Versus Sun Are Mediated by DNA Methylation.

Parental environment influences progeny development in numerous plant and animal systems. Such inherited environmental effects may alter offspring phenotypes in a consistent way, for instance when resource-deprived parents produce low quality offspring due to reduced maternal provisioning. However, because development of individual organisms is guided by both inherited and immediate environmental cues, parental conditions may have different effects depending on progeny environment. Such context-dependent transgenerational plasticity suggests a mechanism of environmental inheritance that can precisely interact with immediate response pathways, such as epigenetic modification. We show that parental light environment (shade versus sun) resulted in context-dependent effects on seedling development in a common annual plant, and that these effects were mediated by DNA methylation. We grew replicate parents of five highly inbred Polygonum persicaria genotypes in glasshouse shade versus sun and, in a fully factorial design, measured ecologically important traits of their isogenic seedling offspring in both environments. Compared to the offspring of sun-grown parents, the offspring of shade-grown parents produced leaves with greater mean and specific leaf area, and had higher total leaf area and biomass. These shade-adaptive effects of parental shade were pronounced and highly significant for seedlings growing in shade, but slight and generally non-significant for seedlings growing in sun. Based on both regression and covariate analysis, inherited effects of parental shade were not mediated by changes to seed provisioning. To test for a role of DNA methylation, we exposed replicate offspring of isogenic shaded and fully insolated parents to either the demethylating agent zebularine or to control conditions during germination, then raised them in simulated growth chamber shade. Partial demethylation of progeny DNA had no phenotypic effect on offspring of shaded parents, but caused offspring of sun-grown parents to develop as if their parents had been shaded, with larger leaves and greater total canopy area and biomass. These results contribute to the increasing body of evidence that DNA methylation can mediate transgenerational environmental effects, and show that such effects may contribute to nuanced developmental interactions between parental and immediate environments.

Developmental plasticity: re-conceiving the genotype.

In recent decades, the phenotype of an organism (i.e. its traits and behaviour) has been studied as the outcome of a developmental 'programme' coded in its genotype. This deterministic view is implicit in the Modern Synthesis approach to adaptive evolution as a sorting process among genetic variants. Studies of developmental pathways have revealed that genotypes are in fact differently expressed depending on environmental conditions. Accordingly, the genotype can be understood as a repertoire of potential developmental outcomes or norm of reaction. Reconceiving the genotype as an environmental response repertoire rather than a fixed developmental programme leads to three critical evolutionary insights. First, plastic responses to specific conditions often comprise functionally appropriate trait adjustments, resulting in an individual-level, developmental mode of adaptive variation. Second, because genotypes are differently expressed depending on the environment, the genetic diversity available to natural selection is itself environmentally contingent. Finally, environmental influences on development can extend across multiple generations via cytoplasmic and epigenetic factors transmitted to progeny individuals, altering their responses to their own, immediate environmental conditions and, in some cases, leading to inherited but non-genetic adaptations. Together, these insights suggest a more nuanced understanding of the genotype and its evolutionary role, as well as a shift in research focus to investigating the complex developmental interactions among genotypes, environments and previous environments.

Invasions and extinctions through the looking glass of evolutionary ecology.

Invasive and endangered species reflect opposite ends of a spectrum of ecological success, yet they experience many similar eco-evolutionary challenges including demographic bottlenecks, hybridization and novel environments. Despite these similarities, important differences exist. Demographic bottlenecks are more transient in invasive species, which (i) maintains ecologically relevant genetic variation, (ii) reduces mutation load, and (iii) increases the efficiency of natural selection relative to genetic drift. Endangered species are less likely to benefit from admixture, which offsets mutation load but also reduces fitness when populations are locally adapted. Invading species generally experience more benign environments with fewer natural enemies, which increases fitness directly and also indirectly by masking inbreeding depression. Adaptive phenotypic plasticity can maintain fitness in novel environments but is more likely to evolve in invasive species encountering variable habitats and to be compromised by demographic factors in endangered species. Placed in an eco-evolutionary context, these differences affect the breadth of the ecological niche, which arises as an emergent property of antagonistic selection and genetic constraints. Comparative studies of invasions and extinctions that apply an eco-evolutionary perspective could provide new insights into the environmental and genetic basis of ecological success in novel environments and improve efforts to preserve global biodiversity.This article is part of the themed issue 'Human influences on evolution, and the ecological and societal consequences'.