The evolutionary ecology of seed germination of Arabidopsis thaliana: variable natural selection on germination timing

Germination timing of Arabidopsis thaliana displays strong plasticity to geographic location and seasonal conditions experienced by seeds. We identified which plastic responses were adaptive using recombinant inbred lines in a field manipulation of geographic location (Kentucky, KY; Rhode Island, RI), maternal photoperiod (14-h and 10-h days), and season of dispersal (June and November). Transgressive segregation created novel genotypes that had either higher fitness or lower fitness in certain environments than either parent. Natural selection on germination timing and its variation explained 72% of the variance in fitness among genotypes in KY, 30% in June-dispersed seeds in RI, but only 4% in November-dispersed seeds in RI. Therefore, natural selection on germination timing is an extremely efficient sieve that can determine which genotypes can persist in some locations, and its efficiency is geographically variable and depends on other aspects of life history. We found no evidence for adaptive responses to maternal photoperiod during seed maturation. We did find adaptive plasticity to season of seed dispersal in RI. Seeds dispersed in June postponed germination, which was adaptive, while seeds dispersed in November accelerated germination, which was also adaptive. We also found maladaptive plasticity to geographic location for seeds dispersed in June, such that seeds dispersed in KY germinated much sooner than the optimum time. Consequently, bet hedging in germination timing was favorable in KY; genotypes with more variation in germination timing had higher fitness because greater variation was associated with postponed germination. Selection on germination timing varied across geographic location, indicating that germination timing can be a critical stage in the establishment of genotypes in new locations. The rate of evolution of germination timing may therefore strongly influence the rate at which species can expand their range.     

The genetics of adaptation to novel environments: selection on germination timing in Arabidopsis thaliana

When studying selection during adaptation to novel environments, researchers have often paid little attention to an organism’s earliest developmental stages. Despite this lack of attention, early life history traits may be under strong selection during colonization, as the expression of adaptive phenotypes at later points is contingent upon early survival. Moreover, the timing of early developmental transitions can constrain the timing of later transitions, with potentially large effects on fitness. In this issue, Huang et al. (2010) underscore the importance of early life history traits in the adaptation of Arabidopsis thaliana to old‐field sites in North America. Using a new population of mapped recombinant inbred lines, the authors examined germination timing and total lifetime fitness of A. thaliana while varying site latitude, dispersal season, and maternal photoperiod. Huang et al. (2010) discovered several Quantitative Trait Loci (QTL) with large effects on fitness that colocalized with QTL for field germination timing and seed dormancy—demonstrating that fitness is genetically associated with these early life history traits, and that these loci are likely under strong selection during adaptation to novel environments. In the epistatic interactions of some loci, recombinant genotypes outperformed parental genotypes, supporting the potentially adaptive role of recombination. This study provides elegant evidence that traits expressed early in an organism’s development can play an important role during adaptive evolution.     

The earliest stages of adaptation in an experimental plant population: strong selection on QTLS for seed dormancy

Colonizing species may often encounter strong selection during the initial stages of adaptation to novel environments. Such selection is particularly likely to act on traits expressed early in development since early survival is necessary for the expression of adaptive phenotypes later in life. Genetic studies of fitness under field conditions, however, seldom include the earliest developmental stages. Using a new set of recombinant inbred lines, we present a study of the genetic basis of fitness variation in Arabidopsis thaliana in which genotypes, environments, and geographic location were manipulated to study total lifetime fitness, beginning with the seed stage. Large‐effect quantitative trait loci (QTLs) for fitness changed allele frequency and closely approached 90% in some treatments within a single generation. These QTLs colocated with QTLs for germination phenology when seeds were dispersed following a schedule of a typical winter annual, and they were detected in two geographic locations at different latitudes. Epistatically interacting loci affected both fitness and germination in many cases. QTLs for field germination phenology colocated with known QTLs for primary dormancy induction as assessed in laboratory tests, including the candidate genes DOG1 and DOG6. Therefore fitness, germination phenology, and primary dormancy are genetically associated at the level of specific chromosomal regions and candidate loci. Genes associated with the ability to arrest development at early life stages and assess environmental conditions are thereby likely targets of intense natural selection early in the colonization process.     

Effects of primary seed dormancy on lifetime fitness of Arabidopsis thaliana in the field

Background and AimsSeed dormancy determines the environmental niche of plants in seasonal environments, and has consequences for plant performance that potentially go far beyond the seed and seedling stages. In this study, we examined the cascading effects of seed dormancy on the expression of subsequent life-history traits and fitness in the annual herb Arabidopsis thaliana.MethodsWe planted seeds of >200 recombinant inbred lines (RILs) derived from a cross between two locally adapted populations (Italy and Sweden), and both parental genotypes at the native site of the Swedish population in three consecutive years. We quantified the relationship between primary seed dormancy and the expression of subsequent life-history traits and fitness in the RIL population with path analysis. To examine the effects of differences in dormancy on the relative fitness of the two parental genotypes, we planted dormant seeds during the seed dispersal period and non-dormant seeds during the germination period of the local population.Key ResultsIn the RIL population, strong primary dormancy was associated with high seedling survival, but with low adult survival and fecundity, and path analysis indicated that this could be explained by effects on germination timing, rosette size and flowering start. The relationship between primary seed dormancy and germination proportion varied among years, and this was associated with differences in seasonal changes in soil moisture. The planting of dormant and non-dormant seeds indicated that the lower primary dormancy of the local Swedish genotype contributed to its higher germination proportion in two years and to its higher fecundity in one year.ConclusionsOur results show that seed dormancy affects trait expression and fitness components across the life cycle, and suggest that among-year variation in the incidence of drought during the germination period should be considered when predicting the consequences of climatic change for population growth and evolution.     

Conflicting selection on the timing of germination in a natural population of Arabidopsis thaliana

The timing of germination is a key life‐history trait that may strongly influence plant fitness and that sets the stage for selection on traits expressed later in the life cycle. In seasonal environments, the period favourable for germination and the total length of the growing season are limited. The optimal timing of germination may therefore be governed by conflicting selection through survival and fecundity. We conducted a field experiment to examine the effects of timing of germination on survival, fecundity and overall fitness in a natural population of the annual herb Arabidopsis thaliana in north‐central Sweden. Seedlings were transplanted at three different times in late summer and in autumn covering the period of seed germination in the study population. Early germination was associated with low seedling survival, but also with high survival and fecundity among established plants. The advantages of germinating early more than balanced the disadvantage and selection favoured early germination. The results suggest that low survival among early germinating seeds is the main force opposing the evolution of earlier germination and that the optimal timing of germination should vary in space and time as a function of the direction and strength of selection acting during different life‐history stages.     

PLEIOTROPY IN THE WILD: THE DORMANCY GENE DOG1 EXERTS CASCADING CONTROL ON LIFE CYCLES

In the wild, organismal life cycles occur within seasonal cycles, so shifts in the timing of developmental transitions can alter the seasonal environment experienced subsequently. Effects of genes that control the timing of prior developmental events can therefore be magnified in the wild because they determine seasonal conditions experienced by subsequent life stages, which can influence subsequent phenotypic expression. We examined such environmentally induced pleiotropy of developmental‐timing genes in a field experiment with Arabidopsis thaliana. When studied in the field under natural seasonal variation, an A. thaliana seed‐dormancy gene, Delay Of Germination 1 (DOG1), was found to influence not only germination, but also flowering time, overall life history, and fitness. Flowering time of the previous generation, in turn, imposed maternal effects that altered germination, the effects of DOG1 alleles, and the direction of natural selection on these alleles. Thus under natural conditions, germination genes act as flowering genes and potentially vice versa. These results illustrate how seasonal environmental variation can alter pleiotropic effects of developmental‐timing genes, such that effects of genes that regulate prior life stages ramify to influence subsequent life stages. In this case, one gene acting at the seed stage impacted the entire life cycle.     

Rainfall seasonality predicts the germination behavior of a tropical dry‐forest vine

Seed dormancy is considered to be an adaptive strategy in seasonal and/or unpredictable environments because it prevents germination during climatically favorable periods that are too short for seedling establishment. Tropical dry forests are seasonal environments where seed dormancy may play an important role in plant resilience and resistance to changing precipitation patterns. We studied the germination behavior of seeds from six populations of the Neotropical vine Dalechampia scandens (Euphorbiaceae) originating from environments of contrasting rainfall seasonality. Seeds produced by second greenhouse‐generation plants were measured and exposed to a favorable wet environment at different time intervals after capsule dehiscence and seed dispersal. We recorded the success and the timing of germination. All populations produced at least some dormant seeds, but seeds of populations originating from more seasonal environments required longer periods of after‐ripening before germinating. Within populations, larger seeds tended to require longer after‐ripening periods than did smaller seeds. These results indicate among‐population genetic differences in germination behavior and suggest that these populations are adapted to local environmental conditions. They also suggest that seed size may influence germination timing within populations. Ongoing changes in seasonality patterns in tropical dry forests may impose strong selection on these traits.     

Effects of changes in the timing and duration of the wet season on the germination of the soil seed bank of a seeder-dominated Mediterranean shrubland

In seasonal climates, rainfall patterns are highly variable across years, and can control seed bank dependent regeneration. Here we asked how changing the timing and duration of the wet season would affect the germination of the soil seed bank of a 14-year-old seeder-dominated shrubland. Soil samples, subjected or not to a heat shock, simulating fire, were set to germinate in a chamber whose conditions (temperature and photoperiod) were successively changed to simulate autumn, winter, and spring. Irrigation was implemented to produce three wet season treatments, varying its timing and duration: long (14?weeks of irrigation, during autumn, winter, and spring), medium (8?weeks, late autumn to early spring) and short (4?weeks in winter). Wet season treatments significantly affected germination of shrubs and herbs, as well as species richness and diversity, whereby the later and shorter the season, the lower these variables. Dicots were more sensitive to the treatments than monocots. The timing of the wet season was also important, as similar significant differences were found when only the first 4 weeks of each simulated wet season treatment were considered; the later the season, the lower the germination and richness. Heating the soil generally increased germination but few significant effects were found. We document that a change in the timing and/or duration of the wet season can significantly affect soil seed bank germination. We discuss these results in a context of shifting rainfall patterns under climate change.     

Precocious floral initiation and identification of exact timing of embryo physiological maturity facilitate germination of immature seeds to truncate the lifecycle of pea

We propose herein a novel single seed descent protocol that has application across a broad phenotypic range of pea genotypes. Manipulation of key in vivo growing conditions, including light, photoperiod and temperature, combined with precocious in vitro germination of the embryo at full physiological maturity substantially shortened the pea lifecycle. We define full embryo physiological maturity as the earliest point in seed development when precocious in vitro germination and robust seedling growth can be reliably achieved without supply of exogenous hormones. Under our optimised conditions for accelerated plant growth, embryo physiological maturity was attained at c. 18 days after pollination, when seed moisture content was below 60?% and sucrose level under 100 mg g^?1 DW. No delay penalty in terms of time to flowering and plant development was caused by the culture of immature seeds 18 days after pollination compared to the used of mature ones. Determining the role embryo maturity plays in the fitness of the germinated plant has facilitated the truncation of the lifecycle across pea genotypes. The accelerated single seed descent system proposed within this research will benefit complex genetic studies via the rapid development of recombinant inbred lines (RIL) and multi-parental advanced generation intercrosses (MAGIC) populations.     

Bigger is not always better: conflicting selective pressures on seed size in Quercus ilex

Most theoretical treatments of the evolutionary ecology of offspring size assume a simple and direct effect of investment per offspring on offspring fitness. In this paper I experimentally determine the relationship between seed mass and several main fitness components of the oak Quercus ilex, to estimate phenotypic selection acting on seed mass during the early life cycle and to discover any potential selective conflicts occurring between different stages from dispersal to establishment. I found a positive effect of acorn size on most fitness components related to seedling establishment. Large size increased germination rate and seedling survival, accelerated germination timing, and enhanced seedling growth. Nevertheless, there was also a direct negative effect of acorn size on survival to predation, because large acorns were highly preferred by the main postdispersal seed predators at the study site, wild boars and wood mice. Because of the low probability of escape from predation, the fitness of large acorns estimated on this component was significantly lower than the fitness of smaller acorns. Therefore, seed size affected fitness in two different ways, yielding opposing and conflicting selective forces. These findings suggest that the general assumption that offspring fitness is a fixed positive function of seed size needs to be reconsidered for some systems. The existence of conflicting selection might explain the occurrence of an optimal seed size in some plant species without invoking a seed number-size trade-off.     

Niche construction through germination cueing: life-history responses to timing of germination in Arabidopsis thaliana

Germination responses to seasonal conditions determine the environment experienced by postgermination life stages, and this ability has potential consequences for the evolution of plant life histories. Using recombinant inbred lines of Arabidopsis thaliana, we tested whether life-history characters exhibited plasticity to germination timing, whether germination timing influenced the strength and mode of natural selection on life-history traits, and whether germination timing influenced the expression of genetic variation for life-history traits. Adult life-history traits exhibited strong plasticity to season of germination, and season of germination significantly altered the strength, mode, and even direction of selection on life-history traits under some conditions. None of the average plastic responses to season of germination or season of dispersal were adaptive, although some genotypes within our sample did exhibit adaptive responses. Thus, recombination between inbred lineages created some novel adaptive genotypes with improved responses to the seasonal timing of germination under some, but not all, conditions. Genetically based variation in germination time tended to augment genetic variances of adult life-history traits, but it did not increase the heritabilities because it also increased environmentally induced variance. Under some conditions, plasticity of life-history traits in response to genetically variable germination timing actually obscured genetic variation for those traits. Therefore, the evolution of germination responses can influence the evolution of life histories in a general manner by altering natural selection on life-history traits and the genetic variation of these traits.     

The effect of maternal and paternal environments on seed characters in the herbaceous plant Campanula Americana (Campanulaceae)

Maternal environments typically influence the phenotype of their offspring. However, the effect of the paternal environment or the potential for joint effects of both parental environments on offspring characters is poorly understood. Two populations of Campanula americana, a woodland herb with a variable life history, were used to determine the influence of maternal and paternal light and nutrient environments on offspring seed characters. Families were grown in the greenhouse in three levels of light or three levels of nutrients. Crosses were conducted within each environmental gradient to produce seeds with all combinations of maternal and paternal environments. On average, increasing maternal nutrient and light levels increased seed mass and decreased percentage germination. The paternal environment affected seed mass, germination time, and percentage germination. However, the influence of the paternal environment varied across maternal environments, suggesting that paternal environmental effects should be evaluated in the context of maternal environments. Significant interactions between family and the parental environments for offspring characters suggest that parental environmental effects are genetically variable. In C. americana, the timing of germination determines life history. Therefore parental environmental effects on germination timing, and genetic variation in those parental effects, suggest that parental environments may influence life history evolution in this system.     

Postdispersal Seed Fate of Tropical Montane Trees in an Agricultural Landscape, Southern Costa Rica

Variation in postdispersal seed fate is an important factor driving patterns of forest regeneration. Because most previous studies have not tracked final seed fate and have commonly equated seed removal with predation without considering the possibility of secondary dispersal, little is known about individual seed mortality factors in successional and degraded habitats. This study tracked the postdispersal fate of large-seeded tropical montane trees in abandoned pasture, young secondary forests, and small forest fragments during two consecutive years in an agricultural landscape in southern Costa Rica. The incidence of secondary dispersal by animals, scatterhoarding in particular, and the effects of seed burial on germination were measured. Overall, seeds survived through germination more often in secondary forests with high levels of mortality occurring in abandoned pastures and forest fragments. The majority of seed mortality was caused by rodent predation in forest fragments, insects and fungal pathogens in secondary forests, and a combination of desiccation, insects, and fungal pathogens in pastures. Seeds were frequently secondarily dispersed in larger forest fragments, whereas they were only rarely moved in pastures and secondary forests. Burial tended to improve germination in pastures and was important for an often scatterhoarded species, Otoba novogranatensis, in all habitats. The results of this study suggest that: (1) seed mortality factors differ in response to the type of habitat degradation; (2) large-seeded species have high potential for survival when dispersed to young secondary forests; and (3) seed removal is not a reliable proxy for seed predation, particularly in less degraded forest fragments.     

ENVIRONMENTAL CONTROL OF PITHOPHORA OEDOGONIA (CHLOROPHYCEAE) AKINETE GERMINATION1

Abundance of Pithophora oedogonia akinetes displayed seasonal changes, being greatest in winter and lowest in summer. Akinete abundance showed significant (P < 0.001) negative correlations with photoperiod(r = -0.53) and water temperature (r= -0.75) during the period February 1978 through June 1979. Experiments in which akinete germination was studied in response to manipulations of nutrients (NO₃-N and PO₄-P), photoperiod and temperature indicated that temperature was the primary factor regulating the timing of the vernal flush of akinete germination observed in Surrey Lake.     

Manipulation of photoperiod perception advances gonadal growth but not laying date in the great tit

In seasonal environments, organisms use biotic and abiotic cues to time various biological processes that are crucial for growth, survival and reproductive success. Photoperiod is the best‐known cue used to regulate gonadal development, migration and moult of many animal species. In birds, the relationship between photoperiod and gonadal development is clearly established, but we have little understanding on whether photoperiod also regulates actual timing of egg laying under natural conditions. Elucidating the link between photoperiod and timing of breeding is however key to understand whether an evolutionary change in sensitivity to photoperiod is a possible mechanism through which organisms could adjust their seasonal timing in response to climate warming. Here, we investigated the causal relationship between photoperiod, gonadal growth and laying date in wild female great tits. We experimentally increased the photoperiod perceived by the birds in spring by clipping head feathers, and we subsequently monitored gonadal development in the lab and egg laying dates in the wild. We show that our manipulation increased the photoperiod perceived by the birds to a level that approximately corresponds to an advancement of ten calendar days. This increase in perceived photoperiod led to an acceleration of gonadal development, but not to an advancement of egg laying dates. Our results indicate that photoperiod sensitivity is not constraining the advancement of laying date under current environmental conditions and suggest that evolution of sensitivity to other supplementary cues is necessary to advance reproduction under global warming.     

Fluctuating natural selection accounts for the evolution of diversification bet hedging

Natural environments are characterized by unpredictability over all time scales. This stochasticity is expected on theoretical grounds to result in the evolution of ‘bet-hedging’ traits that maximize the long term, or geometric mean fitness even though such traits do not maximize fitness over shorter time scales. The geometric mean principle is thus central to our interpretation of optimality and adaptation; however, quantitative empirical support for bet hedging is lacking. Here, I report a quantitative test using the timing of seed germination—a model diversification bet-hedging trait—in Lobelia inflata under field conditions. In a phenotypic manipulation study, I find the magnitude of fluctuating selection acting on seed germination timing—across 70 intervals throughout five seasons—to be extreme: fitness functions for survival are complex and multimodal within seasons and significantly dissimilar among seasons. I confirm that the observed magnitude of fluctuating selection is sufficient to account for the degree of diversification behaviour characteristic of individuals of this species. The geometric mean principle has been known to economic theory for over two centuries; this study now provides a quantitative test of optimality of a bet-hedging trait in nature.     

GERMINATION ECOPHYSIOLOGY OF ARENARIA GLABRA,A WINTER ANNUAL OF SANDSTONE AND GRANITE OUTCROPS OF SOUTHEASTERN UNITED STATES

The germination ecophysiology of Arenaria glabra Michx., a characteristic winter annual plant species of granite and sandstone outcrops of southeastern United States, was investigated. Seeds germinate in early autumn, plants overwinter in the rosette stage and then flower, set seeds, and die in late spring; seeds are dispersed soon after maturity. Eighty-five to 90% of freshly-matured seeds were innately dormant, and the other 10–15% germinated only at temperatures lower than those that occur in the habitat at the time of seed dispersal in June. During the summer after-ripening period, seeds stored dry under ambient laboratory conditions exhibited progressive increases in rates and total percentages of germination, a widening of the temperature range for germination, and a loss of the light requirement. At a 14-hr daily photoperiod, seeds kept on continuously moist soil germinated to 83% at simulated July and August temperatures during July and August, and the remainder germinated at September temperatures in September. On the other hand, seeds subjected to alternate wetting and drying during July and August germinated to only 9% during those 2 months, and the remainder germinated after the soil was kept continuously moist, beginning on 1 September, at simulated habitat temperatures during September and October. Thus, the timing of germination of A. glabra in the field is controlled by an interplay of the seeds' physiological state with the dynamics of temperature and soil moisture conditions.     

Variation in seed traits of Lobelia inflata (Campanulaceae): sources and fitness consequences

Seed germination constitutes an important event in the life cycle of plants. Two related seed traits affect fitness: seed size and the timing of seed germination. In three sets of experiments, we (1) partition the sources of seed-size variance in Lobelia inflata into components attributable to fruit size, relative fruit position, and parental identity; (2) examine the influence of pregermination conditions and seed size on time to germination; and (3) assess the fitness consequences of seed size and germination timing under seminatural, harsh conditions. Seed-size variance is attributable to both parental identity and fruit position within an individual. Distal fruits produce larger but fewer seeds. No significant correlation exists between fruit size and seed size, but a trade-off is found between the number and size of seeds contained in a fruit after correcting for fruit size. The timing of germination is influenced by seed size, light conditions before winter, and winter duration. Germination timing influences survival, and despite small seed size in this species (2 × 10 g/seed), seed size has a persistent and significant association with both final plant size and the probability of survival to autumn.     

Implications of Behavior and Gut Passage for Seed Dispersal Quality: The Case of Black and Gold Howler Monkeys

The most generalized mechanism of seed dispersal in tropical and subtropical forests is dispersal by vertebrates. The pattern of seed dispersal and germination capacity of dispersed seeds can determine the seed disperser quality because both affect the postdispersal fate of dispersed seeds. Animals' behavior is an important determinant of seed deposition pattern. One of most abundant frugivores of Neotropical forests are howler monkeys Alouatta spp., so my aim was to evaluate the potential quality of Alouatta caraya as a seed disperser, obtaining details of: (1) seed deposition pattern, (2) behavioral context of defecations, and (3) germination capacity of dispersed seeds. During 2 years the seed shadow created by howlers and the behavioral context of depositions was examined in the Paraná flooded forest, and germination tests were conducted in the laboratory. Black and gold howler monkeys consumed fruits of five canopy trees, three understory trees, one shrub, and two vines. Howlers dispersed seeds in a complex pattern: 35 percent (337 scats) of scats were deposited individually, and 65 percent were deposited in big latrines associated with the main sleeping trees and in small latrines associated with secondary sleeping trees and confrontations at territory edges. From 261 dispersal distances recorded, 20 and 40 m were the mode, but 57 percent of distances were >50 m and 31 percent were >100 m. Germination speed increased significantly in ingested seeds of Banara arguta, Ocotea diospyrifolia and Eugenia punicifolia . The seed deposition pattern generated by A. caraya may produce a diversification of environmental conditions for dispersed seeds that should be considered in future evaluations.     

A new role for phytochromes in temperature-dependent germination

Germination timing is a fundamental life-history trait, as seedling establishment predicates realized fitness in the wild. Light and temperature are two important cues by which seeds sense the proper season of germination. Using Arabidopsis thaliana, we provide evidence that phytochrome-mediated germination pathways simultaneously respond to light and temperature cues in ways that affect germination. Phytochrome mutant seeds were sown on agar plates and allowed to germinate in lit, growth chambers across a range of temperatures (7 degrees C to 28 degrees C). phyA had an important role in promoting germination at warmer temperatures, phyE was important to germination at colder temperatures and phyB was important to germination across a range of temperatures. Different phytochromes were required for germination at different temperatures, indicating a restriction or even a potential specialization of individual phytochrome activity as a function of temperature. This temperature-dependent activity of particular phytochromes reveals a potentially novel role for phytochrome pathways in regulating the seasonal timing of germination.