Experimental life-history evolution: selection on the allocation to sexual reproduction and its plasticity in a clonal plant

Abstract.— Allocation to sexual reproduction is an important life-history trait in clonal plants. Different selection pressures between competitive and competition-free environments are likely to result in the evolution of specialized genotypes and to maintain genetic variation in reproductive allocation. Moreover, selection may also result in the evolution of plastic allocation strategies. The necessary prerequisite for evolution, heritable genetic variation, can best be studied with selection experiments. Starting from a base population of 102 replicated genotypes of the clonal herb Ranunculus reptans , we imposed selection on the proportion of flowering rosettes in the absence of competition (base population: mean = 0.391, broad-sense heritability = 0.307). We also selected on the plasticity in this trait in response to competition with a naturally coexisting grass in a parallel experiment (base population: 14% lower mean in the presence of competition, broad-sense heritability = 0.072). After two generations of bidirectional selection, the proportion of flowering rosettes was 26% higher in the high line than in the low line (realized heritability ± SE = 0.205 ± 0.017). Moreover, genotypes of the high line had 11% fewer carpels per flower, a 22% lower proportion of rooted rosettes, and a 39% smaller average distance between rosettes within a clone. In the second experiment, we found no significant responses to selection for high and low plasticity in the proportion of flowering rosettes (realized heritability ± SE =–0.002 ± 0.013). Our study indicates a high heritability and potential for further evolution of the proportion of flowering rosettes in R. reptans , but not for its plasticity, which may have been fixed by past evolution at its current level. Moreover, our results demonstrate strong genetic correlations between allocation to sexual reproduction and other clonal life-history characteristics.     

Experimental life-history evolution: selection on growth form and its plasticity in a clonal plant

The growth form along the continuum from compact phalanx plants to more loosely packed guerilla plants is an important life-history trait in clonal plants. Prerequisite for its evolution is heritable genetic variation. Starting with 102 genotypes of the stoloniferous herb Ranunculus reptans, we performed one selection experiment on spatial spread per rosette as measure of guerillaness (broad-sense heritability 0.198) and another on plasticity in this trait in response to competition (broad-sense heritability 0.067). After two generations, spatial spread was 36.9% higher in the high line than in the low line (realized heritability +/- SE 0.149 +/- 0.039). Moreover, compared with the low line genotypes of the high line had fewer rosettes, a lower proportion of flowering rosettes, a higher proportion of rooted rosettes, more branches per rosette, longer internodes and longer leaves. In the second experiment, we found no significant direct response to selection for high and low plasticity in spatial spread (realized heritability +/- SE -0.029 +/- 0.063), despite a significant correlated response in plasticity in the length of the first three stolon internodes. Our study indicates a high potential for further evolution of the clonal growth form in R. reptans, but not for its plasticity, and it demonstrates that the clonal growth form does not evolve independently of other clonal life-history characteristics.     

RAPD variation among and within small and large populations of the rare clonal plant Ranunculus reptans (Ranunculaceae)

In the pre-alpine region of Europe numbers and sizes of populations of the clonal lake shore plant Ranunculus reptans have declined because of the regulation of lake water levels. We investigated genetic variation among and within 17 populations of different size (cover 1–10 000 m²) in R. reptans with RAPD (random amplified polymorphic DNA) profiles. We sampled 127 rosettes in 14 populations at Lake Constance and three populations at or near Lake Como. There was significant genetic variation between plants from the two lake regions (5.9%, analysis of molecular variance [AMOVA], P < 0.001), among populations within lake regions (20.4%, P < 0.001), and within populations (73.7%, P < 0.001). Under the assumptions of Wright's island model the variation among populations corresponds to a gene flow of N_em = 0.70. Within the 14 Lake Constance populations we detected significant genetic variation among subpopulations separated by only a few metres (4.0% of the within-population variation; P < 0.05). Molecular variance was 24% smaller in small populations covering <100 m² area than in larger ones (P < 0.03), indicating that samples from large populations were genetically more variable than samples representing comparable areas of smaller populations. We conclude that gene flow among populations is very limited and that genetic drift has caused reduced genetic variability of smaller populations. Conservation of genetic variability in R. reptans requires persistence of large and also of small populations (because of population differentiation), and it could be enhanced by increasing the size of small populations (to counter genetic drift).     

Selection on phenotypic plasticity of morphological traits in response to flooding and competition in the clonal shore plant Ranunculus reptans

Adaptive evolution of phenotypic plasticity requires that plastic genotypes have the highest global fitness. We studied selection by spatial heterogeneity of interspecific competition and flooding, and by temporal heterogeneity of flooding on morphological plasticity of 52 genotypes of the clonal shore plant Ranunculus reptans. Competition reduced clone size, rosette size, leaf length and stolon internode thickness. Flooding had similar effects and reduced competition. Differences in selection between environments imply potential for either local adaptation or for indirect evolution of phenotypic plasticity. We also detected direct selection for plastic reductions in internode length in response to flooding and for a plastic increase in internode length in response to competition. Plastic responses of some morphological traits to flooding were in line with selection thereon, suggesting that they indeed are adaptive and might have evolved in response to direct selection on plasticity.     

Effects of four generations of density-dependent selection on life history traits and their plasticity in a clonally propagated plant

Life history evolution of many clonal plants takes place with long periods of exclusively clonal reproduction and under largely varying ramet densities resulting from clonal reproduction. We asked whether life history traits of the clonal herb Ranunculus reptans respond to density-dependent selection, and whether plasticity in these traits is adaptive. After four generations of exclusively clonal propagation of 16 low and 16 high ramet-density lines, we studied life history traits and their plasticities at two test ramet-densities. Plastic responses to higher test-density consisted of a shift from sexual to vegetative reproduction, and reduced flower production, plant size, branching frequency, and lengths of leaves and internodes. Plants of high-density lines tended to have longer leaves, and under high test-density branched less frequently than those of low-density lines. Directions of these selection responses indicate that the observed plastic branching response is adaptive, whereas the plastic leaf length response is not. The reverse branching frequency pattern at low test-density, where plants of high-density lines branched more frequently than those of low-density lines, indicates evolution of plasticity in branching. Moreover, when grown under less stressful low test-density, plants of high-density lines tended to grow larger than the ones of low-density lines. We conclude that ramet density affects clonal life-history evolution and that under exclusively clonal propagation clonal life-history traits and their plasticities evolve differently at different ramet densities.     

Constraints on the evolution of phenotypic plasticity in the clonal plant Hydrocotyle vulgaris

The evolution of phenotypic plasticity of plant traits may be constrained by costs and limits. However, the precise constraints are still unclear for many traits under different ecological contexts. In a glasshouse experiment, we grew ramets of 12 genotypes of a clonal plant Hydrocotyle vulgaris under the control (full light and no flood), shade and flood conditions and tested the potential costs and limits of plasticity in 13 morphological and physiological traits in response to light availability and flood variation. In particular, we used multiple regression and correlation analyses to evaluate potential plasticity costs, developmental instability costs and developmental range limits of each trait. We detected significant costs of plasticity in specific petiole length and specific leaf area in response to shade under the full light condition and developmental range limits in specific internode length and intercellular CO₂ concentration in response to light availability variation. However, we did not observe significant costs or limits of plasticity in any of the 13 traits in response to flood variation. Our results suggest that the evolution of phenotypic plasticity in plant traits can be constrained by costs and limits, but such constraints may be infrequent and differ under different environmental contexts.     

植物养分捕获的菌根塑性——外生菌根的塑性

植物根资源捕获塑性是地下生态学研究的重点之一, 在过去二三十年间有长足的进步。菌根塑性是根资源捕获塑性的重要方面, 但由于研究手段的限制, 目前仅有概念上的探讨。缺乏菌根塑性的根塑性研究至少是不全面的。菌根生物学的迅速发展, 尤其是分子生物学手段的介入, 使对菌根塑性进行深入研究成为可能。该文对外生菌根塑性进行讨论, 在简要介绍了外生菌根的生物学基本知识后, 着重讨论了外生菌根形态塑性和生理塑性的定义与内涵。通过文献综述, 分析讨论了外生菌根塑性的研究现状: 很少有研究聚焦在菌根塑性本身, 现有的材料多为其他研究的隐示或研究结果的引申, 并多在形态塑性方面。外生菌根的生理塑性未见有直接的实验数据。该文还对外生菌根研究中发展的、可用于菌根塑性研究的方法进行了综述。由于外生菌根塑性的复杂性, 对菌根塑性的研究会较植物根本身塑性的研究复杂得多, 问题也会相对复杂, 比如植物和外生菌根菌之间的营养需求关系、植物外生菌根塑性的生态意义、实验方法的缺陷等等。对今后外生菌根塑性研究的方向进行了探讨。     

Field evidence of plastic growth responses to habitat heterogeneity in the clonal herbRanunculus repens

Stolon internode lengths were measured on plants of the clonal herbRanunculus repens growing in a hay meadow which was subject to disturbance by mole (Talpa europaea) activity. Within the site three habitat types were recognized: closed grassland, the open ground of fresh molehills and the grass-molehill boundary. The lengths of stolon internodes ofR. repens differed significantly in each of the three habitats. The shortest internodes occurred on stolons on the open molchills. The longest occurred in the closed grassland habitat. The type of habitat in which parent ramets were rooted did not significantly influence the length of internodes on their daughter stolons. The length of a stolon internode was determined by its immediate surrounding habitat type. Consecutive internode lengths on a given stolon showed considerable plasticity, shortening significantly as stolons spread onto molehills from surrounding habitats, and increasing significantly as stolons advanced from a molehill into the surrounding closed grassland habitat. These results are consistent with the proposition that under favorable conditions (on the molehills, where resources are expected to be more abundant, and competition absent) internode lengths shorten and the plant forages intensively, whereas under conditions of low resource availability (in the closed grassland, where competition occurs) internode lengths increase, allowingR. repens to forage extensively. Such morphological plasticity may promote more efficient exploitation of resource-rich sites and more rapid vacation of resource-poor sites.     

Spatial models of foraging in clonal plant species

Ramets of some clonal plant species alter their internode lengths or their frequency of lateral branching in response to their immediate microenvironment. Such “plant foraging” responses are thought to allow clones to concentrate in favorable portions of their environment. Despite widespread interest among ecologists in plant foraging, few realistic models have been developed to examine conditions under which plant foraging responses are likely to provide clones with ecological benefit. In this paper, we develop spatially explicit, stochastic simulation models to examine consequences of both empirical and hypothetical plant foraging responses. We construct a hierarchical series of models in which we incorporate effects of resource heterogeneity on spacer lengths, angles of growth, and lateral branch production. We also vary the number, size, and arrangement of patches, and the presence or absence of ramet mortality. Simulations based on hypothetical data demonstrated the potential importance of shortening spacer lengths in favorable habitat. In these simulations, ramet crowding increased significantly, implying a potential cost to plant foraging responses whose magnitude is large enough to cause ramets to concentrate in favorable patches. Models calibrated with empirical data suggest that when clonal plants were able to concentrate in favorable habitat, this was usually caused by increased daughter ramet production in the favorable habitat. Variation in clonal growth angles had little impact on the ability of ramets or clones to locate favorable patches, but did increase the ability of clones to remain in favorable patches once found. Alterations in the number and size of patches strongly influenced the effectiveness of the foraging response. The spatial arrangement of patches also was important: clumped distributions of patches decreased the success with which plants located favorable patches, especially at the genet level and when the number of patches was low. Finally, when ramet mortality varied with patch quality, there was an increase in the percentage of ramets located in favorable patches; differential ramet mortality also lessened the impact of other effects, such as the decreased success of clones when patches are clumped. Overall, our models indicate that the effectiveness of plant foraging responses is variable and is likely to depend on a suite of environmental conditions.     

Variation in the degree and costs of adaptive phenotypic plasticity among Rana temporaria populations

Adaptive phenotypic plasticity in the form of capacity to accelerate development as a response to pond drying risk is known from many amphibian species. However, very little is known about factors that might constrain the evolution of this type of plasticity, and few studies have explored to what degree plasticity might be constrained by trade-offs dictated by adaptation to different environmental conditions. We compared the ability of southern and northern Scandinavian common frog (Rana temporaria) larvae originating from 10 different populations to accelerate their development in response to simulated pond drying risk and the resulting costs in metamorphic size in a factorial laboratory experiment. We found that (i) northern larvae developed faster than the southern larvae in all treatments, (ii) a capacity to accelerate the response was present in all five southern and all five northern populations tested, but that the magnitude of the response was much larger (and less variable) in the southern than in the northern populations, and that (iii) significant plasticity costs in metamorphic size were present in the southern populations, the plastic genotypes having smaller metamorphic size in the absence of desiccation risk, but no evidence for plasticity costs was found in the northern populations. We suggest that the weaker response to pond drying risk in the northern populations is due to stronger selection on large metamorphic size as compared with southern populations. In other words, seasonal time constraints that have selected the northern larvae to be fast growing and developing, may also constrain their innate ability for adaptive phenotypic plasticity.     

Spatial structure of stoloniferous herbs: an interplay between structural blue-print, ontogeny and phenotypic plasticity

Plant form and spatial structure reflect the basic architectural blue-print of a plant. In most plant species, the expression of the structural blue-print is systematically altered during ontogeny resulting in predictable changes in the allometry of plant structures and in the types of structures that are produced. The expression of the structural blue-print or the timing of ontogenetic changes is also frequently altered by environmental conditions. This latter source of variability, referred to as phenotypic plasticity, is manifested through changes in the timing and rates of meristem initiation and development, the likelihood that meristems will remain dormant or commit to different demographic fates (i.e., vegetative vs. reproductive structures), or the size and structure of the organs formed. Complex interactions among these components can result in considerable differences in form and spatial structure among individuals of the same species. This paper focuses on the importance of these different components in determining the architecture of clonal plants with long internode connections between ramets.A case study is presented that attempts to separate ontogenetic variation and phenotypic plasticity in two stoloniferous species with different structural blue-prints, in their responses to shading. In both species the rate of ontogenetic development responded to intermediate shading levels, but only at very low levels of light availability did plastic changes in branch formation occur. Under shaded conditions the two species achieved similar changes in their architecture in conspicuously different ways. We discuss how different mechanisms leading to a given architecture can be distinguished and what the ecological implications of this are.     

A heuristic model on the role of plasticity in adaptive evolution: plasticity increases adaptation,population viability and genetic variation

An ongoing new synthesis in evolutionary theory is expanding our view of the sources of heritable variation beyond point mutations of fixed phenotypic effects to include environmentally sensitive changes in gene regulation. This expansion of the paradigm is necessary given ample evidence for a heritable ability to alter gene expression in response to environmental cues. In consequence, single genotypes are often capable of adaptively expressing different phenotypes in different environments, i.e. are adaptively plastic. We present an individual-based heuristic model to compare the adaptive dynamics of populations composed of plastic or non-plastic genotypes under a wide range of scenarios where we modify environmental variation, mutation rate and costs of plasticity. The model shows that adaptive plasticity contributes to the maintenance of genetic variation within populations, reduces bottlenecks when facing rapid environmental changes and confers an overall faster rate of adaptation. In fluctuating environments, plasticity is favoured by selection and maintained in the population. However, if the environment stabilizes and costs of plasticity are high, plasticity is reduced by selection, leading to genetic assimilation, which could result in species diversification. More broadly, our model shows that adaptive plasticity is a common consequence of selection under environmental heterogeneity, and hence a potentially common phenomenon in nature. Thus, taking adaptive plasticity into account substantially extends our view of adaptive evolution.     

Habitat selection in spatially heterogeneous environments: a test of foraging behaviour in the clonal submerged macrophyte Vallisneria spiralis

1. To test whether clonal macrophytes can select favourable habitats in heterogeneous environments, clonal fragments of the stoloniferous submerged macrophyte Vallisneria spiralis were subjected to conditions in which light intensity and substratum nutrients were patchily distributed. The allocation of biomass accumulation and ramet production of clones to the different patches was examined. 2. The proportion of both biomass and ramet number of clones allocated to rich patches was significantly higher than in poor patches. The greatest values of both clone and leaf biomass were produced in the heterogeneous light treatment, in which clones originally grew from light‐rich to light‐poor patches, while clones produced the most offspring ramets in the treatments with heterogeneous substratum nutrients. Similarly, root biomass had the highest values in nutrient‐rich patches when clones grew from nutrient‐rich to nutrient‐poor patches. 3. The quality of patches in which parent ramets established significantly influenced the foraging pattern. When previously established in rich patches, a higher proportion of biomass was allocated to rich patches, whereas a higher proportion of ramet number was allocated to rich patches when previously established in poor patches. 4. Results demonstrate that the clonal macrophyte V. spiralis can exhibit foraging in submerged heterogeneous environments: when established under resource‐rich conditions V. spiralis remained in favourable patches, whereas if established in adverse conditions it could escape by allocating more ramets to favourable patches.     

Genetic Allee effects on performance, plasticity and developmental stability in a clonal plant

Negative effects of small population size on fitness, so-called Allee effects, may threaten population persistence even in intact habitat remnants. We studied genotypes of 14 isolated populations of the clonal plant Ranunculus reptans, for which molecular genetic (RAPD-) variability is higher for large than for small populations. In a competition-free greenhouse environment vegetative offspring of genotypes from large populations produced more rosettes and flowers, indicating higher fitness. Within-genotype coefficients of variation in performance traits, indicating developmental instability, were lower for genotypes from populations with higher RAPD-variability. In competition with a taller grass, we found relative reduction in leaf length less pronounced for plants from large populations, suggesting higher adaptive plasticity. Our experimental study of a plant with predominantly vegetative reproduction suggests, that negative genetic effects of recent habitat fragmentation, which so far rather were expected in plants with frequent sexual reproduction, are more severe and more common than previously acknowledged.     

On the evolution of clonal plant life histories

Clonal plant life histories are special in at least four respects: (1) Clonal plants can also reproduce vegetatively, (2) vegetative reproduction can be realised with short or long spacers, (3) and it may allow to plastically place vegetative offspring in benign patches. (4) Moreover, ramets of clonal plants may remain physically and physiologically integrated. Because of the apparent utility of such traits and because ecological patterns of distribution of clonal and non-clonal plants differ, adaptation is a tempting explanation of observed clonal life-history variation. However, adaptive evolution requires (1) heritable genetic variation and (2) a trait effect on fitness, and (3) it may be constrained if other evolutionary forces are overriding selection or by constraints, costs and trade-offs. (1) The few studies undertaken so far reported broad-sense heritability for clonal traits. Variation in selectively neutral genetic markers appears as pronounced in populations of clonal as non-clonal plants. However, neutral markers may not reflect heritable variation of life-history traits. Moreover, clonal plants may have been sampled at larger spatial scales. Empirical information on the contribution of somatic mutations to heritable variation is lacking. (2) Clonal life-history traits were found to affect fitness. However, much of this evidence stems from artificial rather than natural environments. (3) The relative importance of gene flow, inbreeding, and genetic drift, compared with selection, in the evolution of clonal life histories is hardly explored. Benefits of clonal life-history traits were frequently studied and found. However, there is also evidence for constraints, trade-offs, and costs. In conclusion, though it is very likely, that clonal life-history traits are adaptive, it is neither clear to which degree this is the case, nor which clonal life-history traits constitute adaptations to which environmental factors. Moreover, evolutionary interactions among clonal life-history traits and between clonal and non-clonal ones, such as the mating system, are not well explored. There remains much interesting work to be done in this field – which will be particularly interesting if it is done in the field.     

The role of developmental plasticity in evolutionary innovation

Explaining the origins of novel traits is central to evolutionary biology. Longstanding theory suggests that developmental plasticity, the ability of an individual to modify its development in response to environmental conditions, might facilitate the evolution of novel traits. Yet whether and how such developmental flexibility promotes innovations that persist over evolutionary time remains unclear. Here, we examine three distinct ways by which developmental plasticity can promote evolutionary innovation. First, we show how the process of genetic accommodation provides a feasible and possibly common avenue by which environmentally induced phenotypes can become subject to heritable modification. Second, we posit that the developmental underpinnings of plasticity increase the degrees of freedom by which environmental and genetic factors influence ontogeny, thereby diversifying targets for evolutionary processes to act on and increasing opportunities for the construction of novel, functional and potentially adaptive phenotypes. Finally, we examine the developmental genetic architectures of environment-dependent trait expression, and highlight their specific implications for the evolutionary origin of novel traits. We critically review the empirical evidence supporting each of these processes, and propose future experiments and tests that would further illuminate the interplay between environmental factors, condition-dependent development, and the initiation and elaboration of novel phenotypes.     

Evolution of phenotypic plasticity: Genetic differentiation and additive genetic variation for induced plant defence in wild arugula Eruca sativa

Phenotypic plasticity is the primary mechanism of organismal resilience to abiotic and biotic stress, and genetic differentiation in plasticity can evolve if stresses differ among populations. Inducible defence is a common form of adaptive phenotypic plasticity, and long‐standing theory predicts that its evolution is shaped by costs of the defensive traits, costs of plasticity and a trade‐off in allocation to constitutive versus induced traits. We used a common garden to study the evolution of defence in two native populations of wild arugula Eruca sativa (Brassicaceae) from contrasting desert and Mediterranean habitats that differ in attack by caterpillars and aphids. We report genetic differentiation and additive genetic variance for phenology, growth and three defensive traits (toxic glucosinolates, anti‐nutritive protease inhibitors and physical trichome barriers) as well their inducibility in response to the plant hormone jasmonic acid. The two populations were strongly differentiated for plasticity in nearly all traits. There was little evidence for costs of defence or plasticity, but constitutive and induced traits showed a consistent additive genetic trade‐off within each population for the three defensive traits. We conclude that these populations have evolutionarily diverged in inducible defence and retain ample potential for the future evolution of phenotypic plasticity in defence.     

Effects of clonal integration on plant plasticity in Fragaria chiloensis

The ability of clonal plants to transport substances between ramets located in different microsites also allows them to modify the plastic responses of individual ramets to local environmental conditions. By equalising concentrations of substances between ramets, physiological integration might decrease responses to local conditions. However, integration has also been observed to increase plasticity and induce novel plastic responses in ramets. To ask how integration modifies plant plasticity in the clonal herb, Fragaria chiloensis, ramets were given either low light and high nitrogen or high light and low nitrogen, simulating a pattern of resource patchiness in their native habitat. Ramets in contrasting light/nitrogen treatments were either connected or single. Effects of light/nitrogen and connection were measured at three levels of morphological organisation, the organ, the ramet, and the clonal fragment. Connection between ramets reduced or had no effect on plastic responses in leaf size at the level of the plant organ. This suggested that integration dampened certain plastic responses. Connection induced a new plastic response at the level of the clonal fragment, an increase in allocation to vegetative reproduction in patches of low light and high nitrogen. It is concluded that clonal integration can have different effects on plant plasticity at different levels of plant organisation. It appears that, at least in this species, integration can increase plasticity at the level of the clonal fragment and concentrate vegetative reproduction in particular microsite types.     

Extreme growth plasticity in the early branching sauropodomorph Massospondylus carinatus

There is growing evidence of developmental plasticity in early branching dinosaurs and their outgroups. This is reflected in disparate patterns of morphological and histological change during ontogeny. In fossils, only the osteohistological assessment of annual lines of arrested growth (LAGs) can reveal the pace of skeletal growth. Some later branching non-bird dinosaur species appear to have followed an asymptotic growth pattern, with declining growth rates at increasing ontogenetic ages. By contrast, the early branching sauropodomorph Plateosaurus trossingensis appears to have had plastic growth, suggesting that this was the plesiomorphic condition for dinosaurs. The South African sauropodomorph Massospondylus carinatus is an ideal taxon in which to test this because it is known from a comprehensive ontogenetic series, it has recently been stratigraphically and taxonomically revised, and it lived at a time of ecosystem upheaval following the end-Triassic extinction. Here, we report on the results of a femoral osteohistological study of M. carinatus comprising 20 individuals ranging from embryo to skeletally mature. We find major variability in the spacing of the LAGs and infer disparate body masses for M. carinatus individuals at given ontogenetic ages, contradicting previous studies. These findings are consistent with a high degree of growth plasticity in M. carinatus.