The chicken or the egg? Adaptation to desiccation and salinity tolerance in a lineage of water beetles

Transitions from fresh to saline habitats are restricted to a handful of insect lineages, as the colonization of saline waters requires specialized mechanisms to deal with osmotic stress. Previous studies have suggested that tolerance to salinity and desiccation could be mechanistically and evolutionarily linked, but the temporal sequence of these adaptations is not well established for individual lineages. We combined molecular, physiological and ecological data to explore the evolution of desiccation resistance, hyporegulation ability (i.e., the ability to osmoregulate in hyperosmotic media) and habitat transitions in the water beetle genus Enochrus subgenus Lumetus (Hydrophilidae). We tested whether enhanced desiccation resistance evolved before increases in hyporegulation ability or vice versa, or whether the two mechanisms evolved in parallel. The most recent ancestor of Lumetus was inferred to have high desiccation resistance and moderate hyporegulation ability. There were repeated shifts between habitats with differing levels of salinity in the radiation of the group, those to the most saline habitats generally occurring more rapidly than those to less saline ones. Significant and accelerated changes in hyporegulation ability evolved in parallel with smaller and more progressive increases in desiccation resistance across the phylogeny, associated with the colonization of meso‐ and hypersaline waters during global aridification events. All species with high hyporegulation ability were also desiccation‐resistant, but not vice versa. Overall, results are consistent with the hypothesis that desiccation resistance mechanisms evolved first and provided the physiological basis for the development of hyporegulation ability, allowing these insects to colonize and diversify across meso‐ and hypersaline habitats.     

Reduced salinities compromise the thermal tolerance of hypersaline specialist diving beetles

Inland saline waters are globally threatened habitats that harbour unique assemblages of specialist invertebrates. In many Mediterranean regions, irrigation associated with intensive agriculture is lowering the salinity of these habitats, resulting in the loss of their specialist biota, although the mechanisms by which reductions in salinity lead to species loss are poorly understood. In the present study, the effects of reduced salinity on the temperature tolerance and thermal acclimatory abilities of two related species of hypersaline water beetles, Nebrioporus baeticus (Schaum) and Nebrioporus ceresyi (Aubé), are explored. Both upper (UTL) and lower thermal limits (LTL) are assessed, and both salinity and temperature are found to influence the thermal biology of Nebrioporus. Mean UTLs are greater in individuals of both species acclimated at high salinities, with salinity appearing to be more important than acclimation temperature in determining UTL. In both taxa, the lowest mean LTLs are recorded in individuals acclimated at the highest salinities and lowest temperatures; temperature‐dependent acclimation is only reported after exposure to relatively high salinities. The data show that salinity influences the thermal tolerance and acclimatory ability of these hypersaline beetles, and that lowered salinity compromises the ability of adult Nebrioporus to cope with both heat and cold. Such an effect may partly explain why specialist species are lost from hypersaline habitats subject to salinity reductions, and suggests that ongoing reduction in salinity may compromise the ability of such specialist taxa to cope with rapid climate change.     

Tempo and mode of the multiple origins of salinity tolerance in a water beetle lineage

Salinity is one of the most important drivers of the distribution, abundance and diversity of organisms. Previous studies on the evolution of saline tolerance have been mainly centred on marine and terrestrial organisms, while lineages inhabiting inland waters remain largely unexplored. This is despite the fact that these systems include a much broader range of salinities, going from freshwater to more than six times the salinity of the sea (i.e. >200 g/L). Here, we study the pattern and timing of the evolution of the tolerance to salinity in an inland aquatic lineage of water beetles (Enochrus species of the subgenus Lumetus, family Hydrophilidae), with the general aim of understanding the mechanisms by which it was achieved. Using a time‐calibrated phylogeny built from five mitochondrial and two nuclear genes and information about the salinity tolerance and geographical distribution of the species, we found that salinity tolerance appeared multiple times associated with periods of global aridification. We found evidence of some accelerated transitions from freshwater directly to high salinities, as reconstructed with extant lineages. This, together with the strong positive correlation found between salinity tolerance and aridity of the habitats in which species are found, suggests that tolerance to salinity may be based on a co‐opted mechanism developed originally for drought resistance.     

Characterization of halotolerant Bicosoecida and Placididea (Stramenopila) that are distinct from marine forms,and the phylogenetic pattern of salinity preference in heterotrophic stramenopiles

Recent culture‐based studies demonstrate the distinctiveness of the microbial eukaryote biota of very hypersaline environments. In contrast, microscopy‐based faunistic studies suggest that the biota of habitats of more moderate hypersalinity (60–150‰) overlaps substantially with that of marine environments, but this has barely been studied with modern techniques. To investigate the diversity and salinity tolerance range of these organisms, eight cultures of heterotrophic stramenopiles were established from (or from nearby) moderately hypersaline locations. These isolates represent five independent groups; Groups A, B and C are bicosoecids; Groups D and E belong to Placididea. One isolate (Group A) is a strain of the widespread marine species Cafeteria roenbergensis, and cannot grow above 100‰ salinity. The other isolates – Groups B–E – can all grow at 150–175‰ salinities and are probably moderate halophiles. Groups B–E all represent previously unsequenced species or even genera, although Group B is the sister group of the borderline extreme halophile Halocafeteria. The high level of novelty en countered suggests that moderately hypersaline environments may harbour a heterotrophic stramenopile biota distinct from that of marine environments. Interestingly, our new isolates are all most closely related to marine or halophilic forms, and our phylogenies show large clades defined by saline/non‐saline habitats within bicosoecids, placidomonads and related lineages. In particular, most freshwater/soil bicosoecids form one well‐supported clade. The sole major exception is Bicosoeca, which is intermixed with marine environmental sequences originally referred to as ‘MAST‐13’, which are from brackish water, not typical seawater. It seems that the freshwater/marine barrier has been crossed very few times in the evolutionary history of these heterotrophic stramenopile flagellates.     

Gradients of salinity stress, environmental stability and water chemistry as a templet for defining habitat types and physiological strategies in inland salt waters

The search for pattern in the geographic occurrence of salt lake flora and fauna often reveals strong associations of specific taxa with certain types of water chemistry. Solute composition, along with salinity and habitat stability, may provide a templet shaping the distribution of many organisms inhabiting saline lakes. A review of studies demonstrating habitat associations, specific solute tolerance, and ionic and osmotic adaptations provide evidence of fidelity to particular conditions of environmental chemistry across a wide taxonomic spectrum. Under low salinity conditions, some species show osmoregulatory adaptability to varied solute composition but the capacity for such flexibility is reduced with increased salinity and only certain taxa are found in hypersaline waters dominated by a particular solute. Anionic ratios of chloride, bicarbonate–carbonate, and sulfate appear to be especially important determinants of distribution. Specific solute tolerance presents an alternative explanation to disrupted hydrographic connections in describing how biogeographic distributions may be restricted to certain aquatic habitats in arid regions. Physiological adaptations to chemistry, exemplified in the brine fly genus Ephydra, may be an integral part of the evolution, ecology and diversification of saline water organisms.     

Salt‐tolerance diversity in diploid and polyploid cotton (Gossypium) species

The development of salt‐tolerant genotypes is pivotal for the effective utilization of salinized land and to increase global crop productivity. Several cotton species comprise the most important source of textile fibers globally, and these are increasingly grown on marginal or increasingly saline agroecosystems. The allopolyploid cotton species also provide a model system for polyploid research, of relevance here because polyploidy was suggested to be associated with increased adaptation to stress. To evaluate the genetic variation of salt tolerance among cotton species, 17 diverse accessions of allopolyploid (AD‐genome) and diploid (A‐ and D‐genome) Gossypium were evaluated for a total of 29 morphological and physiological traits associated with salt tolerance. For most morphological and physiological traits, cotton accessions showed highly variable responses to 2 weeks of exposure to moderate (50 mm NaCl) and high (100 mm NaCl) hydroponic salinity treatments. Our results showed that the most salt‐tolerant species were the allopolyploid Gossypium mustelinum from north‐east Brazil, the D‐genome diploid Gossypium klotzschianum from the Galapagos Islands, followed by the A‐genome diploids of Africa and Asia. Generally, A‐genome accessions outperformed D‐genome cottons under salinity conditions. Allopolyploid accessions from either diploid genomic group did not show significant differences in salt tolerance, but they were more similar to one of the two progenitor lineages. Our findings demonstrate that allopolyploidy in itself need not be associated with increased salinity stress tolerance and provide information for using the secondary Gossypium gene pool to breed for improved salt tolerance.     

Is salinity an obstacle for biological invasions?

Invasions of freshwater habitats by marine and brackish species have become more frequent in recent years with many of those species originating from the Ponto‐Caspian region. Populations of Ponto‐Caspian species have successfully established in the North and Baltic Seas and their adjoining rivers, as well as in the Great Lakes–St. Lawrence River region. To determine if Ponto‐Caspian taxa more readily acclimatize to and colonize diverse salinity habitats than taxa from other regions, we conducted laboratory experiments on 22 populations of eight gammarid species native to the Ponto‐Caspian, Northern European and Great Lakes–St. Lawrence River regions. In addition, we conducted a literature search to survey salinity ranges of these species worldwide. Finally, to explore evolutionary relationships among examined species and their populations, we sequenced the mitochondrial cytochrome c oxidase subunit I gene (COI) from individuals used for our experiments. Our study revealed that all tested populations tolerate wide ranges of salinity, however, different patterns arose among species from different regions. Ponto‐Caspian taxa showed lower mortality in fresh water, while Northern European taxa showed lower mortality in fully marine conditions. Genetic analyses showed evolutionary divergence among species from different regions. Due to the geological history of the two regions, as well as high tolerance of Ponto‐Caspian species to fresh water, whereas Northern European species are more tolerant of fully marine conditions, we suggest that species originating from the Ponto‐Caspian and Northern European regions may be adapted to freshwater and marine environments, respectively. Consequently, the perception that Ponto‐Caspian species are more successful colonizers might be biased by the fact that areas with highest introduction frequency of NIS (i.e., shipping ports) are environmentally variable habitats which often include freshwater conditions that cannot be tolerated by euryhaline taxa of marine origin.     

Opposing patterns of zooplankton diversity and functioning along a natural stress gradient: when the going gets tough,the tough get going

Salinity represents a major structuring factor in aquatic habitats which strongly affects species richness. We studied the relationships among species richness, density and phylogenetic diversity of zooplankton communities along a natural salinity gradient in astatic soda pans in the Carpathian Basin (Hungary, Austria and Serbia). Diversity and density showed opposing trends along the salinity gradient. The most saline habitats had communities of one or two species only, with maximum densities well above 1000 ind l^?1. Similarity of communities increased with salinity, with most of the highly saline communities being dominated by one highly tolerant calanoid copepod, Arctodiaptomus spinosus, which was at the same time the only soda‐water specialist. Salinity obviously constrained species composition and resulted in communities of low complexity, where few tolerant species ensure high biomass production in the absence of antagonistic interactions. The pattern suggests that environmental stress may result in highly constrained systems which exhibit high rates of functioning due to these key species, in spite of the very limited species pool.     

The Comparative Osmoregulatory Ability of Two Water Beetle Genera Whose Species Span the Fresh-Hypersaline Gradient in Inland Waters (Coleoptera: Dytiscidae,Hydrophilidae)

A better knowledge of the physiological basis of salinity tolerance is essential to understanding the ecology and evolutionary history of organisms that have colonized inland saline waters. Coleoptera are amongst the most diverse macroinvertebrates in inland waters, including saline habitats; however, the osmoregulatory strategies they employ to deal with osmotic stress remain unexplored. Survival and haemolymph osmotic concentration at different salinities were examined in adults of eight aquatic beetle species which inhabit different parts of the fresh—hypersaline gradient. Studied species belong to two unrelated genera which have invaded saline waters independently from freshwater ancestors; Nebrioporus (Dytiscidae) and Enochrus (Hydrophilidae). Their osmoregulatory strategy (osmoconformity or osmoregulation) was identified and osmotic capacity (the osmotic gradient between the animal’s haemolymph and the external medium) was compared between species pairs co-habiting similar salinities in nature. We show that osmoregulatory capacity, rather than osmoconformity, has evolved independently in these different lineages. All species hyperegulated their haemolymph osmotic concentration in diluted waters; those living in fresh or low-salinity waters were unable to hyporegulate and survive in hyperosmotic media (> 340 mosmol kg^-1). In contrast, the species which inhabit the hypo-hypersaline habitats were effective hyporegulators, maintaining their haemolymph osmolality within narrow limits (ca. 300 mosmol kg^-1) across a wide range of external concentrations. The hypersaline species N. ceresyi and E. jesusarribasi tolerated conductivities up to 140 and 180 mS cm^-1, respectively, and maintained osmotic gradients over 3500 mosmol kg^-1, comparable to those of the most effective insect osmoregulators known to date. Syntopic species of both genera showed similar osmotic capacities and in general, osmotic responses correlated well with upper salinity levels occupied by individual species in nature. Therefore, osmoregulatory capacity may mediate habitat segregation amongst congeners across the salinity gradient.     

Salt tolerance evolves more frequently in C4 grass lineages

Salt tolerance has evolved many times in the grass family, and yet few cereal crops are salt tolerant. Why has it been so difficult to develop crops tolerant of saline soils when salt tolerance has evolved so frequently in nature? One possible explanation is that some grass lineages have traits that predispose them to developing salt tolerance and that without these background traits, salt tolerance is harder to achieve. One candidate background trait is photosynthetic pathway, which has also been remarkably labile in grasses. At least 22 independent origins of the C₄ photosynthetic pathway have been suggested to occur within the grass family. It is possible that the evolution of C₄ photosynthesis aids exploitation of saline environments, because it reduces transpiration, increases water‐use efficiency and limits the uptake of toxic ions. But the observed link between the evolution of C₄ photosynthesis and salt tolerance could simply be due to biases in phylogenetic distribution of halophytes or C₄ species. Here, we use a phylogenetic analysis to investigate the association between photosynthetic pathway and salt tolerance in the grass family Poaceae. We find that salt tolerance is significantly more likely to occur in lineages with C₄ photosynthesis than in C₃ lineages. We discuss the possible links between C₄ photosynthesis and salt tolerance and consider the limitations of inferring the direction of causality of this relationship.     

Transcriptomic differences between euryhaline and stenohaline malaria vector sibling species in response to salinity stress

Evolution of osmoregulatory systems is a key factor in the transition of species between fresh‐ and saltwater habitats. Anopheles coluzzii and Anopheles merus are stenohaline and euryhaline malaria vector mosquitoes belonging to a larger group of sibling species, the Anopheles gambiae complex, which radiated in Africa within the last 2 million years. Comparative ecological genomics of these vector species can provide insight into the mechanisms that permitted the rapid radiation of this species complex into habitats of contrasting salinity. Here, we use RNA‐Seq to investigate gene expression differences between An. coluzzii and An. merus after briefly exposing both young and old larval instars of each species to either saltwater (SW) or freshwater (FW). Our study aims to identify candidate genes and pathways responsible for the greater SW tolerance of An. merus. Our results are congruent with the ability of gene induction to mediate salinity tolerance, with both species showing increasing amounts of differential gene expression between SW and FW as salt concentrations increase. Besides ion transporters such as AgAE2 that may serve as effectors for osmoregulation, we also find mitogen‐activated protein kinases that may serve in a phosphorylation signalling pathway responding to salinity, and report potential cross‐talk between the mosquito immune response and osmoregulation. This study provides a key step towards applying the growing molecular knowledge of these malaria vectors to improve understanding of their ecological tolerances and habitat occupancy.     

Phenotypic plasticity of four Chenopodiaceae species with contrasting saline–sodic tolerance in response to increased salinity–sodicity

It is unknown whether phenotypic plasticity in fitness‐related traits is associated with salinity–sodicity tolerance. This study compared growth and allocation phenotypic plasticity in two species with low salinity–sodicity tolerance (Chenopodium acuminatum and C. stenophyllum) and two species with high salinity–sodicity tolerance (Suaeda glauca and S. salsa) in a pot experiment in the Songnen grassland, China. While the species with low tolerance had higher growth and allocation plasticity than the highly tolerant species, the highly tolerant species only adjusted their growth traits and maintained higher fitness (e.g., plant height and total biomass) in response to increased soil salinity–sodicity, with low biomass allocation plasticity. Most plasticity is “apparent” plasticity (ontogenetic change), and only a few traits, for example, plant height:stem diameter ratio and root:shoot biomass ratio, represent “real” plasticity (real change in response to the environment). Our results show that phenotypic plasticity was negatively correlated with saline–sodic tolerance and could be used as an index of species sensitivity to soil salinity–sodicity.     

A molecular phylogeny of the north‐east Atlantic species of the genus Idotea (Isopoda) with focus on the Baltic Sea

Today, the marine isopods of the genus Idotea Fabricius, 1798 consist of 26 accepted species. Most species can be found in the Northern Hemisphere. While some species have a cosmopolitan distribution, others are endemic to a few coastlines. In the Baltic Sea, Idotea species have a key role as important meso‐grazers. Only three species can be found in this brackish environment, I. balthica, I. chelipes and I. granulosa, while nine species are described from the whole European coast. The goal of the present study was to reconstruct the phylogeny of the genus Idotea and to test whether the Baltic low‐salinity tolerant species represent a single lineage within the genus. The phylogeny of north‐east Atlantic Idotea species was investigated based on a fragment of the nuclear gene 28S and the mitochondrial gene COI for eight Idotea species. The phylogenetic reconstructions showed that the genus Idotea might not be monophyletic. Neither I. linearis nor I. urotoma did belong to the separated and well‐supported Idotea clade of I. balthica, I. chelipes, I. emarginata, I. granulosa, I. metallica and I. pelagica. The three Idotea species found in the Baltic belonged to different lineages, with estimated COI‐based divergence time older than 7 Myr. This suggests that the tolerance to low salinity has likely evolved in the genus Idotea more than once.     

High diversity and isolated distribution of aquatic heterotrophic protists in salars of the Atacama Desert at different salinities

The species richness of eukaryotes in the hypersaline environment is generally thought to be low. However, recent studies showed a high degree of phylogenetic novelty at these extreme conditions with variable chemical parameters. These findings call for a more thorough look into the species richness of hypersaline environments. In this study, various hypersaline lakes (salars, 1–348 PSU) as well as further aquatic ecosystems of northern Chile were investigated regarding diversity of heterotrophic protists by metabarcoding studies of surface water samples. Investigations of genotypes of 18S rRNA genes showed a unique community composition in nearly each salar and even among different microhabitats within one salar. The genotype distribution showed no clear connection to the composition of main ions at the sampling sites, but protist communities from similar salinity ranges (either hypersaline, hyposaline or mesosaline) clustered together regarding their OTU composition. Salars appeared to be fairly isolated systems with only little exchange of protist communities where evolutionary lineages could separately evolve.     

Naturally Saline Boreal Communities as Models for Reclamation of Saline Oil Sand Tailings

Reclaimed landscapes after oil sands mining have saline soils; yet, they are required to have similar biodiversity and productivity as the predisturbance nonsaline landscape. Given that many species in the boreal forest are not tolerant of salinity, we studied the effects of soil salinity on plant communities in natural saline landscapes to understand potential plant responses during the reclamation process. Vegetation–soil relationships were measured along transects from flooded wetlands to upland forest vegetation in strongly saline, slightly saline, nonsaline, and reclaimed boreal landscapes. In strongly saline landscapes, surface soil salinity was high (>10 dS/m) in flooded, wet‐meadow, and dry‐meadow vegetation zones as compared to slightly saline (<5 dS/m) and nonsaline (<2 dS/m) landscapes. Plant communities in these vegetation zones were quite different from nonsaline boreal landscapes and were dominated by halophytes common to saline habitats of the Great Plains. In the shrub and forest vegetation zones, surface soil salinity was similar between saline and nonsaline landscapes, resulting in similar plant communities. In strongly saline landscapes, soils remained saline at depth through the shrub and forest vegetation zones (>10 dS/m), suggesting that forest vegetation can establish over saline soils as long as the salts are below the rooting zone. The reclaimed landscape was intermediate between slightly saline and nonsaline landscapes in terms of soil salinity but more similar to nonsaline habitats with respect to species composition. Results from this study suggest it may be unrealistic to expect that plant communities similar to those found on the predisturbance landscape can be established on all reclaimed landscapes after oil sands mining.     

Morphological and structural plasticity of grassland species in response to a gradient in saline‐sodic soils

The abundance and distribution of species can be ascribed to both environmental heterogeneity and stress tolerance, with the latter measure sometimes associated with phenotypic plasticity. Although phenotypic plasticity varies predictably in response to common forms of stress, we lack a mechanistic understanding of the response of species to high saline‐sodic soils. We compared the phenotypic plasticity of three pairs of high and low saline‐sodic tolerant congeners from the families Poaceae (Leymus chinensis versus L. secalinus), Fabaceae (Lespedeza davurica versus L. bicolor) and Asteraceae (Artemisia mongolica versus A. sieversiana) in a controlled pot experiment in the Songnen grassland, China. The low tolerant species, L. secalinus and A. sieversiana exhibited higher plasticity in response to soil salinity and sodicity than their paired congeners. Highly tolerant species, L. chinensis and A. mongolica, had higher values for several important morphological traits, such as shoot length and total biomass under the high saline‐sodic soil treatment than their paired congeners. In contrast, congeners from the family Fabaceae, L. davurica and L. bicolor, did not exhibit significantly different plasticity in response to soil salinity and sodicity. All species held a constant reproductive effort in response to saline‐sodic soil stress. The different responses between low and high tolerant species offer an explanation for the distribution patterns of these species in the Songnen grassland. Highly tolerant species showed less morphological plasticity over a range of saline‐sodic conditions than their paired congeners, which may manifest as an inability to compete with co‐occurring species in locations where saline‐sodic soils are absent.     

Irreversible habitat specialization does not constrain diversification in hypersaline water beetles

Specialization to extreme environments is often considered an evolutionary dead‐end, leading to irreversible adaptations and reduced evolvability. There is, however, mixed evidence for this macroevolutionary pattern, and limited data from speciose lineages. Here, we tested the effect of habitat specialization to hypersaline waters in the diversification rates of aquatic beetles of the genus Ochthebius (Coleoptera, Hydraenidae), using a molecular phylogeny with more than 50% of the 546 recognized species, including representatives of all but one of the nine recognized subgenera and 17 species groups. Phylogenies were built combining mitochondrial and nuclear genes, with the addition of 42 mitochondrial genomes. Using Bayesian methods of character reconstruction, we show that hypersaline tolerance is an irreversible ecological specialization that arose multiple times. Two lineages of Ochthebius experienced a significant increase in diversification rates, one of them inhabiting hypersaline waters, but there was no overall correlation with habitat or any significant decrease in diversification rates despite the irreversibility of hypersaline tolerance. Our study tested for the first time the impact of hypersaline habitat specialization on diversification rates, finding no support for it to be an evolutionary dead‐end. On the contrary, multiple and ancient lineages fully adapted to these extreme osmotic conditions have persisted and diversified over a long evolutionary timescale.     

Salt- and alkaline-tolerance are linked in Acacia

Saline or alkaline soils present a strong stress on plants that together may be even more deleterious than alone. Australia''s soils are old and contain large, sometimes overlapping, areas of high salt and alkalinity. Acacia and other Australian plant lineages have evolved in this stressful soil environment and present an opportunity to understand the evolution of salt and alkalinity tolerance. We investigate this evolution by predicting the average soil salinity and pH for 503 Acacia species and mapping the response onto a maximum-likelihood phylogeny. We find that salinity and alkalinity tolerance have evolved repeatedly and often together over 25 Ma of the Acacia radiation in Australia. Geographically restricted species are often tolerant of extreme conditions. Distantly related species are sympatric in the most extreme soil environments, suggesting lack of niche saturation. There is strong evidence that many Acacia have distributions affected by salinity and alkalinity and that preference is lineage specific.