CAX1 suppresses Cd‐induced generation of reactive oxygen species in Arabidopsis halleri

The molecular analysis of metal hyperaccumulation in species such as Arabidopsis halleri offers the chance to gain insights into metal homeostasis and into the evolution of adaptation to extreme habitats. A prerequisite of metal hyperaccumulation is metal hypertolerance. Genetic analysis of a backcross population derived from Arabidopsis lyrata × A. halleri crosses revealed three quantitative trait loci for Cd hypertolerance. A candidate gene for Cdtol2 is AhCAX1, encoding a vacuolar Ca²⁺/H⁺ antiporter. We developed a method for the transformation of vegetatively propagated A. halleri plants and generated AhCAX1‐silenced lines. Upon Cd²⁺ exposure, several‐fold higher accumulation of reactive oxygen species (ROS) was detectable in roots of AhCAX1‐silenced plants. In accordance with the dependence of Cdtol2 on external Ca²⁺ concentration, this phenotype was exclusively observed in low Ca²⁺ conditions. The effects of external Ca²⁺ on Cd accumulation cannot explain the phenotype as they were not influenced by the genotype. Our data strongly support the hypothesis that higher expression of CAX1 in A. halleri relative to other Arabidopsis species represents a Cd hypertolerance factor. We propose a function of AhCAX1 in preventing a positive feedback loop of Cd‐elicited ROS production triggering further Ca²⁺‐dependent ROS accumulation.     

Metal hyperaccumulation and hypertolerance: a model for plant evolutionary genomics

In the course of evolution, plants adapted to widely differing metal availabilities in soils and therefore represent an important source of natural variation of metal homeostasis networks. Research on plant metal homeostasis can thus provide insights into the functioning, regulation and adaptation of biological networks. Here, we describe major recent breakthroughs in the understanding of the genetic and molecular basis of metal hyperaccumulation and associated hypertolerance, a naturally selected complex trait which represents an extreme adaptation of the metal homeostasis network. Investigations in this field reveal further the molecular alterations underlying the evolution of natural phenotypic diversity and provide a highly relevant framework for comparative genomics.     

Effects of assay conditions in life history experiments with Drosophila melanogaster

Selection experiments with Drosophila have revealed constraints on the simultaneous evolution of life history traits. However, the responses to selection reported by different research groups have not been consistent. Two possible reasons for these inconsistencies are (i) that different groups used different environments for their experiments and (ii) that the selection environments were not identical to the assay environments in which the life history traits were measured. We tested for the effect of the assay environment in life history experiments by measuring a set of Drosophila selection lines in laboratories working on life history evolution with Drosophila in Basel, Groningen, Irvine and London. The lines measured came from selection experiments from each of these laboratories. In each assay environment, we measured fecundity, longevity, development time and body size. The results show that fecundity measurements were particularly sensitive to the assay environment. Differences between assay and selection environment in the same laboratory or differences between assay environments between laboratories could have contributed to the differences in the published results. The other traits measured were less sensitive to the assay environment. However, for all traits there were cases where the measurements in one laboratory suggested that selection had an effect on the trait, whereas in other laboratories no such conclusion would have been drawn. Moreover, we provide good evidence for local adaptation in early fecundity for lines from two laboratories.     

Protein deprivation facilitates the independent evolution of behavior and morphology

Ecological conditions such as nutrition can change genetic covariances between traits and accelerate or slow down trait evolution. As adaptive trait correlations can become maladaptive following rapid environmental change, poor or stressful environments are expected to weaken genetic covariances, thereby increasing the opportunity for independent evolution of traits. Here, we demonstrate the differences in genetic covariance among multiple behavioral and morphological traits (exploration, aggression, and body weight) between southern field crickets (Gryllus bimaculatus) raised in favorable (free‐choice) versus stressful (protein‐deprived) nutritional environments. We also quantify the extent to which differences in genetic covariance structures contribute to the potential for the independent evolution of these traits. We demonstrate that protein‐deprived environments tend to increase the potential for traits to evolve independently, which is caused by genetic covariances that are significantly weaker for crickets raised on protein‐deprived versus free‐choice diets. The weakening effects of stressful environments on genetic covariances tended to be stronger in males than in females. The weakening of the genetic covariance between traits under stressful nutritional environments was expected to facilitate the opportunity for adaptive evolution across generations. Therefore, the multivariate gene‐by‐environment interactions revealed here may facilitate behavioral and morphological adaptations to rapid environmental change.     

Demographic history of the trace metal hyperaccumulator Noccaea caerulescens (J. Presl and C. Presl) F. K. Mey. in Western Europe

Noccaea caerulescens (Brassicaceae) is a major pseudometallophyte model for the investigation of the genetics and evolution of metal hyperaccumulation in plants. We studied the population genetics and demographic history of this species to advance the understanding of among‐population differences in metal hyperaccumulation and tolerance abilities. Sampling of seven to 30 plants was carried out in 62 sites in Western Europe. Genotyping was carried out using a combination of new chloroplast and nuclear neutral markers. A strong genetic structure was detected, allowing the definition of three genetic subunits. Subunits showed a good geographic coherence. Accordingly, distant metallicolous populations generally belonged to distinct subunits. Approximate Bayesian computation analysis of demographic scenarios among subunits further supported a primary isolation of populations from the southern Massif Central prior to last glacial maximum, whereas northern populations may have derived during postglacial recolonization events. Estimated divergence times among subunits were rather recent in comparison with the species history, but certainly before the establishment of anthropogenic metalliferous sites. Our results suggest that the large‐scale genetic structure of N. caerulescens populations pre‐existed to the local adaptation to metalliferous sites. The population structure of quantitative variation for metal‐related adaptive traits must have established independently in isolated gene pools. However, features of the most divergent genetic unit (e.g. extreme levels of Cd accumulation observed in previous studies) question the putative relationships between adaptive evolution of metal‐related traits and subunits isolation. Finally, admixture signals among distant metallicolous populations suggest a putative role of human activities in facilitating long‐distance genetic exchanges.     

Multiple Novel Traits without Immediate Benefits Originate in Bacteria Evolving on Single Antibiotics

How new traits originate in evolution is a fundamental question of evolutionary biology. When such traits arise, they can either be immediately beneficial in their environment of origin, or they may become beneficial only in a future environment. Compared to immediately beneficial novel traits, novel traits without immediate benefits remain poorly studied. Here we use experimental evolution to study novel traits that are not immediately beneficial but that allow bacteria to survive in new environments. Specifically, we evolved multiple E. coli populations in five antibiotics with different mechanisms of action, and then determined their ability to grow in more than 200 environments that are different from the environment in which they evolved. Our populations evolved viability in multiple environments that contain not just clinically relevant antibiotics, but a broad range of antimicrobial molecules, such as surfactants, organic and inorganic salts, nucleotide analogues and pyridine derivatives. Genome sequencing of multiple evolved clones shows that pleiotropic mutations are important for the origin of these novel traits. Our experiments, which lasted fewer than 250 generations, demonstrate that evolution can readily create an enormous reservoir of latent traits in microbial populations. These traits can facilitate adaptive evolution in a changing world.     

Evolutionary potential in the wild: more than meets the eye

The genus Aquilegia consists of 60–70 perennial plant species widely distributed throughout the northern hemisphere. Its flowers have a delicate and ornamental appearance that makes them a favourite of gardeners. In this genus, adaptive radiations for both floral and vegetative traits have occurred. These adaptive radiations, and the key phylogenetic placement of Aquilegia between Arabidopsis and rice, make this genus a ‘model system’ for plant evolution ( Kramer 2009 ). In this issue, Castellanos et al. (2011) use a marker‐based method to infer heritability for floral and vegetative traits in two Aquilegia species. Layered on top of this are estimates of the strength of natural selection. This novel joint estimation of heritability and selection in the wild showed that vegetative traits, compared to floral traits, have the highest evolutionarily potential. Evolutionary potential is the most important quantity to measure in wild populations. It combines inheritance and strength of selection and predicts the potential for populations to adapt to changing environments. The combination of molecular techniques with species in natural environments makes this work a model for molecular ecological investigations.     

Nickel hyperaccumulation as an anti-herbivore trait: considering the role of tolerance to damage

Metal hyperaccumulation is a striking trait exhibited by many plant species, but the evolutionary ecology of metal hyperaccumulation is poorly understood. It has been widely hypothesized that metal hyperaccumulation evolved to protect plants from herbivory. However, there is currently little evidence that metal hyperaccumulation enhances the fitness of plants in the presence of herbivory. In this study, we conducted a multi-factor greenhouse experiment to examine the effects of two soil nickel concentrations (unamended (0 μg/g) and nickel amended (600 μg/g)), and three levels of artificial damage (0, 10 and 50%) on the growth of plants from two populations of Thlaspi montanum var. montanum. We observed a significant interaction between soil nickel and artificial damage. An a posteriori analysis of this interaction revealed that the presence of nickel significantly improved the ability of T. montanum to tolerate the negative effects of intense damage. Our results indicate that metal hyperaccumulation could benefit T. montanum by increasing its tolerance to damage. This study suggests that there is a potential for the evolution of metal hyperaccumulation in response to intense herbivory on T. montanum.     

Testing drivers of acoustic divergence in cicadas (Cicadidae: Tettigettalna)

Divergence in acoustic signals may have a crucial role in the speciation process of animals that rely on sound for intra-specific recognition and mate attraction. The acoustic adaptation hypothesis (AAH) postulates that signals should diverge according to the physical properties of the signalling environment. To be efficient, signals should maximize transmission and decrease degradation. To test which drivers of divergence exert the most influence in a speciose group of insects, we used a phylogenetic approach to the evolution of acoustic signals in the cicada genus Tettigettalna, investigating the relationship between acoustic traits (and their mode of evolution) and body size, climate and micro-/macro-habitat usage. Different traits showed different evolutionary paths. While acoustic divergence was generally independent of phylogenetic history, some temporal variables’ divergence was associated with genetic drift. We found support for ecological adaptation at the temporal but not the spectral level. Temporal patterns are correlated with micro- and macro-habitat usage and temperature stochasticity in ways that run against the AAH predictions, degrading signals more easily. These traits are likely to have evolved as an anti-predator strategy in conspicuous environments and low-density populations. Our results support a role of ecological selection, not excluding a likely role of sexual selection in the evolution of Tettigettalna calling songs, which should be further investigated in an integrative approach.     

Environmental heterogeneity,multivariate sexual selection and genetic constraints on cuticular hydrocarbons in Drosophila simulans

Sexual selection is responsible for the evolution of many elaborate traits, but sexual trait evolution could be influenced by opposing natural selection as well as genetic constraints. As such, the evolution of sexual traits could depend heavily on the environment if trait expression and attractiveness vary between environments. Here, male Drosophila simulans were reared across a range of diets and temperatures, and we examined differences between these environments in terms of (i) the expression of male cuticular hydrocarbons (CHCs) and (ii) which male CHC profiles were most attractive to females. Temperature had a strong effect on male CHC expression, whereas the effect of diet was weaker. Male CHCs were subject to complex patterns of directional, quadratic and correlational sexual selection, and we found differences between environments in the combination of male CHCs that were most attractive to females, with clearer differences between diets than between temperatures. We also show that genetic covariance between environments is likely to cause a constraint on independent CHC evolution between environments. Our results demonstrate that even across the narrow range of environmental variation studied here, predicting the outcome of sexual selection can be extremely complicated, suggesting that studies ignoring multiple traits or environments may provide an over‐simplified view of the evolution of sexual traits.     

Metal hyperaccumulation: a model system for coevolutionary studies

Recent years have seen a flurry of research activity concerning the hyperaccumulation of heavy metals by plants. Much of the interest in hyperaccumulation has been fueled by the commercial potential of phytoremediation, the use of plants to clean up contaminated soils (Baker et al ., 1994; Salt et al ., 1995; Chaney et al ., 1997, 2000). These applications have in turn spurred many studies of the genetics and physiology of metal uptake (e.g. Krämer et al ., 1996; Lasat et al ., 1996; Salt & Krämer, 1999; Baker et al ., 2000; see also Krämer, 2000; Lombi et al ., 2000). Although phytoremediation provides an intriguing and potentially profitable backdrop, the ecology and evolution of hyperaccumulation in natural populations are interesting subjects in their own right. Two papers in this issue (Ghaderian et al ., pp. 219–224; Davis & Boyd, pp. 211–217) are exciting contributions to the growing consensus that hyperaccumulation may act as a defense against herbivores and pathogens, and suggest that hyperaccumulation may become a model system for research in this area.     

ADAPTATION TO DESICCATION FAILS TO GENERATE PRE‐ AND POSTMATING ISOLATION IN REPLICATE DROSOPHILA MELANOGASTER LABORATORY POPULATIONS

Many laboratory speciation experiments have raised allopatric populations in different environments to determine whether reproductive isolation evolves as a byproduct of adaptation (a form of ecological speciation). Few, however, have addressed the evolution of both pre‐ and postmating isolation or investigated the conditions affecting the process. We present results of an evolution experiment in which 12 lines of Drosophila melanogaster were derived from a common population and then independently evolved for more than 57 generations under alternative selection treatments. Six “desiccation” populations had access to food and water removed during a period of their adult lives generating strong desiccation selection, and six “starvation” populations had access to food but not water removed for the same period, generating a mild starvation stress. Substantial divergence of cuticular hydrocarbons occurred between the desiccation and starvation populations, key traits that have been implicated in sexual isolation in Drosophila. Despite this divergence, there was no detectable premating isolation between desiccation and starvation populations and postmating isolation was asymmetrical: the fitness of F1 hybrids was reduced in the desiccation but not the starvation environment. This asymmetry was likely caused by the absence of divergent selection: adaptation to desiccation appears to have come at no cost to performance in the starvation environment. Novel environments are thought to promote the evolution of reproductive isolation. Understanding the conditions that favor or hamper this remains a key challenge for speciation research.     

Evolutionary dynamics of quantitative variation in an adaptive trait at the regional scale: The case of zinc hyperaccumulation in Arabidopsis halleri

Metal hyperaccumulation in plants is an ecological trait whose biological significance remains debated, in particular because the selective pressures that govern its evolutionary dynamics are complex. One of the possible causes of quantitative variation in hyperaccumulation may be local adaptation to metalliferous soils. Here, we explored the population genetic structure of Arabidopsis halleri at fourteen metalliferous and nonmetalliferous sampling sites in southern Poland. The results were integrated with a quantitative assessment of variation in zinc hyperaccumulation to trace local adaptation. We identified a clear hierarchical structure with two distinct genetic groups at the upper level of clustering. Interestingly, these groups corresponded to different geographic subregions, rather than to ecological types (i.e., metallicolous vs. nonmetallicolous). Also, approximate Bayesian computation analyses suggested that the current distribution of A. halleri in southern Poland could be relictual as a result of habitat fragmentation caused by climatic shifts during the Holocene, rather than due to recent colonization of industrially polluted sites. In addition, we find evidence that some nonmetallicolous lowland populations may have actually derived from metallicolous populations. Meanwhile, the distribution of quantitative variation in zinc hyperaccumulation did separate metallicolous and nonmetallicolous accessions, indicating more recent adaptive evolution and diversifying selection between metalliferous and nonmetalliferous habitats. This suggests that zinc hyperaccumulation evolves both ways—towards higher levels at nonmetalliferous sites and lower levels at metalliferous sites. Our results open a new perspective on possible evolutionary relationships between A. halleri edaphic types that may inspire future genetic studies of quantitative variation in metal hyperaccumulation.     

The genetic architecture of ecological specialization: correlated gene effects on host use and habitat choice in pea aphids

Genetic correlations among phenotypic characters result when two traits are influenced by the same genes or sets of genes. By reducing the degree to which traits in two environments can evolve independently (e.g., Lande 1979; Via and Lande 1985), such correlations are likely to play a central role in both the evolution of ecological specialization and in its link to speciation. For example, negative genetic correlations between fitness traits in different environments (i.e., genetic trade-offs) are thought to influence the evolution of specialization, while positive genetic correlations between performance and characters influencing assortative mating can accelerate the evolution of reproductive isolation between ecologically specialized populations. We first discuss how the genetic architecture of a suite of traits may affect the evolutionary role of genetic correlations among them and review how the mechanisms of correlations can be analyzed using quantitative trait locus (QTL) mapping. We then consider the implications of such data for understanding the evolution of specialization and its link to speciation. We illustrate this approach with a QTL analysis of key characters in two races of pea aphids that are highly specialized on different host plants and partially reproductively isolated. Our results suggest that antagonism among QTL effects on performance in the two environments leads to a genetic trade-off in this system. We also found evidence for parallel QTL effects on host-plant acceptance and fecundity on the accepted host, which could produce assortative mating. These results suggest that the genetic architecture of traits associated with host use may have played a central role in the evolution of specialization and reproductive isolation in pea aphids.     

中国西南四种常见松树火适应性状比较

野火对松属植物的进化和分布有重要的影响。在与野火长期抗争中,松属植物形成了一系列性状提高在易火生境中的适合度,维持种群生存与繁衍。西南地区是中国野火高发区,作为这一地区特有松树,云南松、思茅松和高山松具有一些典型的火适应性状,如厚树皮。以这3种松树和东部常见的马尾松为研究对象,比较4种松树的火适应对策。结果表明：4种松树的火适应性状存在一定的差异。与火适应相关的13项性状主成分分析显示,4个种在空间上总体是分离的,也表现出一些重叠。其中云南松火适应生活史对策是火耐受型和火依赖型的中间类型,适应会发生林冠火的生境。高山松、思茅松和马尾松都是火耐受型,通过快速高生长、厚树皮等性状适应生境不同频度的地表火。四种松树火适应对策与分布区火险基本相符。表明野火在这一区域广泛存在,并对植物进化和森林格局有重要影响。野火在西南地区松属分布和种群维持中的生态作用应被给予足够重视。     

Phytoextraction of metals and metalloids from contaminated soils

The removal of inorganic contaminants by plants is termed phytoextraction. Recent studies have looked at the feasibility of phytoextraction, and demonstrate that both good biomass yields and metal hyperaccumulation are required to make the process efficient. Adding chelating agents to soil to increase the bioavailability of contaminants can sometimes induce hyperaccumulation in normal plants, but may produce undesirable environmental risks. Thus, it is necessary to investigate the mechanisms responsible for hyperaccumulation, using natural hyperaccumulators as model plant species. Recent advances have been made in understanding the mechanisms responsible for hyperaccumulation of Zn, Cd, Ni and As by plants. Attempts to engineer metal tolerance and accumulation have so far been limited to Hg, As and Cd, and although promising results have been obtained they may be some way from practical application. More fundamental understanding of the traits and mechanisms involved in hyperaccumulation are needed so that phytoextraction can be optimised.     

Genetic correlation between temperature-induced plasticity of life-history traits in a soil arthropod

Temperature is considered one of the most important mediators of phenotypic plasticity in ectotherms. However, the costs and benefits shaping the evolution of different thermal responses are poorly elucidated. One of the possible constraints to phenotypic plasticity is its intrinsic genetic cost, such as genetic linkage or pleiotropy. Genetic coupling of the thermal response curves for different life history traits may significantly affect the evolution of thermal sensitivity in thermally fluctuating environments. We used the collembolan Orchesella cincta to study if there is genetic variation in temperature-induced phenotypic plasticity in life history traits, and if the degree of temperature-induced plasticity is correlated across traits. Egg development rate, juvenile growth rate and egg size of 19 inbred isofemale lines were measured at two temperatures. Our results show that temperature was a highly significant factor for all three traits. Egg development rate and juvenile growth rate increased with increasing temperature, while egg size decreased. Line by temperature interaction was significant for all traits tested; indicating that genetic variation for temperature-induced plasticity existed. The degree of plasticity was significantly positively correlated between egg development rate and growth rate, but plasticity in egg size was not correlated to the other two plasticity traits. The findings suggest that the thermal plasticities of egg development rate and growth rate are partly under the control of the same genes or genetic regions. Hence, evolution of the thermal plasticity of traits cannot be understood in isolation of the response of other traits. If traits have similar and additive effects on fitness, genetic coupling between these traits may well facilitate the evolution of optimal phenotypes. However, for this we need to know the selective forces under field conditions.