Environmental RNA degrades more rapidly than environmental DNA across a broad range of pH conditions.

Although the use and development of molecular biomonitoring tools based on environmental nucleic acids (eDNA and eRNA; collectively known as eNAs) have gained broad interest for the quantification of biodiversity in natural ecosystems, studies investigating the impact of site-specific physicochemical parameters on eNA-based detection methods (particularly eRNA) remain scarce. Here, we used a controlled laboratory microcosm experiment to comparatively assess the environmental degradation of eDNA and eRNA across an acid-base gradient following complete removal of the progenitor organism (Daphnia pulex). Using water samples collected over a 30-day period, eDNA and eRNA copy numbers were quantified using a droplet digital PCR (ddPCR) assay targeting the mitochondrial cytochrome c oxidase subunit I (COI) gene of D. pulex. We found that eRNA decayed more rapidly than eDNA at all pH conditions tested, with detectability-predicted by an exponential decay model-for up to 57 h (eRNA; neutral pH) and 143 days (eDNA; acidic pH) post organismal removal. Decay rates for eDNA were significantly higher in neutral and alkaline conditions than in acidic conditions, while decay rates for eRNA did not differ significantly among pH levels. Collectively, our findings provide the basis for a predictive framework assessing the persistence and degradation dynamics of eRNA and eDNA across a range of ecologically relevant pH conditions, establish the potential for eRNA to be used in spatially and temporally sensitive biomonitoring studies (as it is detectable across a range of pH levels), and may be used to inform future sampling strategies in aquatic habitats.

A Glyphosate-Based Herbicide Cross-Selects for Antibiotic Resistance Genes in Bacterioplankton Communities.

Agrochemicals often contaminate freshwater bodies, affecting microbial communities that underlie aquatic food webs. For example, the herbicide glyphosate has the potential to indirectly select for antibiotic-resistant bacteria. Such cross-selection could occur if the same genes (encoding efflux pumps, for example) confer resistance to both glyphosate and antibiotics. To test for cross-resistance in natural aquatic bacterial communities, we added a glyphosate-based herbicide (GBH) to 1,000-liter mesocosms filled with water from a pristine lake. Over 57 days, we tracked changes in bacterial communities with shotgun metagenomic sequencing and annotated metagenome-assembled genomes (MAGs) for the presence of known antibiotic resistance genes (ARGs), plasmids, and resistance mutations in the enzyme targeted by glyphosate (enolpyruvyl-shikimate-3-phosphate synthase; EPSPS). We found that high doses of GBH significantly increased ARG frequency and selected for multidrug efflux pumps in particular. The relative abundance of MAGs after a high dose of GBH was predictable based on the number of ARGs in their genomes (17% of variation explained) and, to a lesser extent, by resistance mutations in EPSPS. Together, these results indicate that GBHs can cross-select for antibiotic resistance in natural freshwater bacteria. IMPORTANCE Glyphosate-based herbicides (GBHs) such as Roundup formulations may have the unintended consequence of selecting for antibiotic resistance genes (ARGs), as demonstrated in previous experiments. However, the effects of GBHs on ARGs remain unknown in natural aquatic communities, which are often contaminated with pesticides from agricultural runoff. Moreover, the resistance provided by ARGs compared to canonical mutations in the glyphosate target enzyme, EPSPS, remains unclear. Here, we performed a freshwater mesocosm experiment showing that a GBH strongly selects for ARGs, particularly multidrug efflux pumps. These selective effects were evident after just a few days, and the ability of bacteria to survive and thrive after GBH stress was predictable by the number of ARGs in their genomes and, to a lesser extent, by mutations in EPSPS. Intensive GBH application may therefore have the unintended consequence of selecting for ARGs in natural freshwater communities.

Warming winters in lakes: Later ice onset promotes consumer overwintering and shapes springtime planktonic food webs.

Global climate warming is causing the loss of freshwater ice around the Northern Hemisphere. Although the timing and duration of ice covers are known to regulate ecological processes in seasonally ice-covered ecosystems, the consequences of shortening winters for freshwater biota are poorly understood owing to the scarcity of under-ice research. Here, we present one of the first in-lake experiments to postpone ice-cover onset (by ≤21 d), thereby extending light availability (by ≤40 d) in early winter, and explicitly demonstrate cascading effects on pelagic food web processes and phenologies. Delaying ice-on elicited a sequence of events from winter to spring: 1) relatively greater densities of algal resources and primary consumers in early winter; 2) an enhanced prevalence of winter-active (overwintering) consumers throughout the ice-covered period, associated with augmented storage of high-quality fats likely due to a longer access to algal resources in early winter; and 3) an altered trophic structure after ice-off, with greater initial springtime densities of overwintering consumers driving stronger, earlier top-down regulation, effectively reducing the spring algal bloom. Increasingly later ice onset may thus promote consumer overwintering, which can confer a competitive advantage on taxa capable of surviving winters upon ice-off; a process that may diminish spring food availability for other consumers, potentially disrupting trophic linkages and energy flow pathways over the subsequent open-water season. In considering a future with warmer winters, these results provide empirical evidence that may help anticipate phenological responses to freshwater ice loss and, more broadly, constitute a case of climate-induced cross-seasonal cascade on realized food web processes.

Widespread agrochemicals differentially affect zooplankton biomass and community structure.

Anthropogenic environmental change is causing habitat deterioration at unprecedented rates in freshwater ecosystems. Despite increasing more rapidly than many other agents of global change, synthetic chemical pollution-including agrochemicals such as pesticides-has received relatively little attention in freshwater community and ecosystem ecology. Determining the combined effects of multiple agrochemicals on complex biological systems remains a major challenge, requiring a cross-field integration of ecology and ecotoxicology. Using a large-scale array of experimental ponds, we investigated the response of zooplankton community properties (biomass, composition, and diversity metrics) to the individual and joint presence of three globally widespread agrochemicals: the herbicide glyphosate, the neonicotinoid insecticide imidacloprid, and nutrient fertilizers. We tracked temporal variation in zooplankton biomass and community structure along single and combined pesticide gradients (each spanning eight levels), under low (mesotrophic) and high (eutrophic) nutrient-enriched conditions, and quantified (1) response threshold concentrations, (2) agrochemical interactions, and (3) community resistance and recovery. We found that the biomass of major zooplankton groups differed in their sensitivity to pesticides: ≥0.3 mg/L glyphosate elicited long-lasting declines in rotifer communities, both pesticides impaired copepods (≥3 µg/L imidacloprid and ≥5.5 mg/L glyphosate), whereas some cladocerans were highly tolerant to pesticide contamination. Strong interactive effects of pesticides were only recorded in ponds treated with the combination of the highest doses. Overall, glyphosate was the most influential driver of aggregate community properties of zooplankton, with biomass and community structure responding rapidly but recovering unequally over time. Total community biomass showed little resistance when first exposed to glyphosate, but rapidly recovered and even increased with glyphosate concentration over time; in contrast, taxon richness decreased in more contaminated ponds but failed to recover. Our results indicate that the biomass of tolerant taxa compensated for the loss of sensitive species after the first exposure, conferring greater community resistance upon a subsequent contamination event; a case of pollution-induced community tolerance in freshwater animals. These findings suggest that zooplankton biomass may be more resilient to agrochemical pollution than community structure; yet all community properties measured in this study were affected at glyphosate concentrations below common water quality guidelines in North America.

Resistance, resilience, and functional redundancy of freshwater bacterioplankton communities facing a gradient of agricultural stressors in a mesocosm experiment.

Agricultural pollution with fertilizers and pesticides is a common disturbance to freshwater biodiversity. Bacterioplankton communities are at the base of aquatic food webs, but their responses to these potentially interacting stressors are rarely explored. To test the extent of resistance and resilience in bacterioplankton communities faced with agricultural stressors, we exposed freshwater mesocosms to single and combined gradients of two commonly used pesticides: the herbicide glyphosate (0-15 mg/L) and the neonicotinoid insecticide imidacloprid (0-60 µg/L), in high or low nutrient backgrounds. Over the 43-day experiment, we tracked variation in bacterial density with flow cytometry, carbon substrate use with Biolog EcoPlates, and taxonomic diversity and composition with environmental 16S rRNA gene amplicon sequencing. We show that only glyphosate (at the highest dose, 15 mg/L), but not imidacloprid, nutrients, or their interactions measurably changed community structure, favouring members of the Proteobacteria including the genus Agrobacterium. However, no change in carbon substrate use was detected throughout, suggesting functional redundancy despite taxonomic changes. We further show that communities are resilient at broad, but not fine taxonomic levels: 24 days after glyphosate application the precise amplicon sequence variants do not return, and tend to be replaced by phylogenetically close taxa. We conclude that high doses of glyphosate - but still within commonly acceptable regulatory guidelines - alter freshwater bacterioplankton by favouring a subset of higher taxonomic units (i.e., genus to phylum) that transiently thrive in the presence of glyphosate. Longer-term impacts of glyphosate at finer taxonomic resolution merit further investigation.

Adaptation and competition in deteriorating environments.

Evolution might rescue populations from extinction in changing environments. Using experimental evolution with microalgae, we investigated if competition influences adaptation to an abiotic stressor, and vice versa, if adaptation to abiotic change influences competition. In a first set of experiments, we propagated monocultures of five species with and without increasing salt stress for approximately 180 generations. When assayed in monoculture, two of the five species showed signatures of adaptation, that is, lines with a history of salt stress had higher population growth rates at high salt than lines without prior exposure to salt. When assayed in mixtures of species, however, only one of these two species had increased population size at high salt, indicating that competition can alter how adaptation to abiotic change influences population dynamics. In a second experiment, we cultivated two species in monocultures and in pairs, with and without increasing salt. While we found no effect of competition on adaptation to salt, our experiment revealed that evolutionary responses to salt can influence competition. Specifically, one of the two species had reduced competitive ability in the no-salt environment after long-term exposure to salt stress. Collectively, our results highlight the complex interplay of adaptation to abiotic change and competitive interactions.

Landscape modification and nutrient-driven instability at a distance.

Almost 50 years ago, Michael Rosenzweig pointed out that nutrient addition can destabilise food webs, leading to loss of species and reduced ecosystem function through the paradox of enrichment. Around the same time, David Tilman demonstrated that increased nutrient loading would also be expected to cause competitive exclusion leading to deleterious changes in food web diversity. While both concepts have greatly illuminated general diversity-stability theory, we currently lack a coherent framework to predict how nutrients influence food web stability across a landscape. This is a vitally important gap in our understanding, given mounting evidence of serious ecological disruption arising from anthropogenic displacement of resources and organisms. Here, we combine contemporary theory on food webs and meta-ecosystems to show that nutrient additions are indeed expected to drive loss in stability and function in human-impacted regions. Our models suggest that destabilisation is more likely to be caused by the complete loss of an equilibrium due to edible plant species being competitively excluded. In highly modified landscapes, spatial nutrient transport theory suggests that such instabilities can be amplified over vast distances from the sites of nutrient addition. Consistent with this theoretical synthesis, the empirical frequency of these distant propagating ecosystem imbalances appears to be growing. This synthesis of theory and empirical data suggests that human modification of the Earth is strongly connecting distantly separated ecosystems, causing rapid, expansive and costly nutrient-driven instabilities over vast areas of the planet. Similar to existing food web theory, the corollary to this spatial nutrient theory is that slowing down spatial nutrient pathways can be a potent means of stabilising degraded ecosystems.

Community rescue in experimental phytoplankton communities facing severe herbicide pollution.

Community rescue occurs when ecological or evolutionary processes restore positive growth in a highly stressful environment that was lethal to the community in its ancestral form, thus averting biomass collapse in a deteriorating environment. Laboratory evidence suggests that community rescue is most likely in high-biomass communities that have previously experienced moderate doses of sublethal stress. We assessed this result under more natural conditions, in a mesocosm experiment with phytoplankton communities exposed to the ubiquitous herbicide glyphosate. We tested whether community biomass and prior herbicide exposure would facilitate community rescue after severe contamination. We found that prior exposure to glyphosate was a very strong predictor of the rescue outcome, while high community biomass was not. Furthermore, although glyphosate had negative effects on diversity, it did not influence community composition significantly, suggesting a modest role for genus sorting in this rescue process. Our results expand the scope of community rescue theory to complex ecosystems and confirm that prior stress exposure is a key predictor of rescue.

Long-term cyclic persistence in an experimental predator-prey system.

Predator-prey cycles rank among the most fundamental concepts in ecology, are predicted by the simplest ecological models and enable, theoretically, the indefinite persistence of predator and prey1-4. However, it remains an open question for how long cyclic dynamics can be self-sustained in real communities. Field observations have been restricted to a few cycle periods5-8 and experimental studies indicate that oscillations may be short-lived without external stabilizing factors9-19. Here we performed microcosm experiments with a planktonic predator-prey system and repeatedly observed oscillatory time series of unprecedented length that persisted for up to around 50 cycles or approximately 300 predator generations. The dominant type of dynamics was characterized by regular, coherent oscillations with a nearly constant predator-prey phase difference. Despite constant experimental conditions, we also observed shorter episodes of irregular, non-coherent oscillations without any significant phase relationship. However, the predator-prey system showed a strong tendency to return to the dominant dynamical regime with a defined phase relationship. A mathematical model suggests that stochasticity is probably responsible for the reversible shift from coherent to non-coherent oscillations, a notion that was supported by experiments with external forcing by pulsed nutrient supply. Our findings empirically demonstrate the potential for infinite persistence of predator and prey populations in a cyclic dynamic regime that shows resilience in the presence of stochastic events.

Understanding Maladaptation by Uniting Ecological and Evolutionary Perspectives.

Evolutionary biologists have long trained their sights on adaptation, focusing on the power of natural selection to produce relative fitness advantages while often ignoring changes in absolute fitness. Ecologists generally have taken a different tack, focusing on changes in abundance and ranges that reflect absolute fitness while often ignoring relative fitness. Uniting these perspectives, we articulate various causes of relative and absolute maladaptation and review numerous examples of their occurrence. This review indicates that maladaptation is reasonably common from both perspectives, yet often in contrasting ways. That is, maladaptation can appear strong from a relative fitness perspective, yet populations can be growing in abundance. Conversely, resident individuals can appear locally adapted (relative to nonresident individuals) yet be declining in abundance. Understanding and interpreting these disconnects between relative and absolute maladaptation, as well as the cases of agreement, is increasingly critical in the face of accelerating human-mediated environmental change. We therefore present a framework for studying maladaptation, focusing in particular on the relationship between absolute and relative fitness, thereby drawing together evolutionary and ecological perspectives. The unification of these ecological and evolutionary perspectives has the potential to bring together previously disjunct research areas while addressing key conceptual issues and specific practical problems.

Causes of maladaptation.

Evolutionary biologists tend to approach the study of the natural world within a framework of adaptation, inspired perhaps by the power of natural selection to produce fitness advantages that drive population persistence and biological diversity. In contrast, evolution has rarely been studied through the lens of adaptation's complement, maladaptation. This contrast is surprising because maladaptation is a prevalent feature of evolution: population trait values are rarely distributed optimally; local populations often have lower fitness than imported ones; populations decline; and local and global extinctions are common. Yet we lack a general framework for understanding maladaptation; for instance in terms of distribution, severity, and dynamics. Similar uncertainties apply to the causes of maladaptation. We suggest that incorporating maladaptation-based perspectives into evolutionary biology would facilitate better understanding of the natural world. Approaches within a maladaptation framework might be especially profitable in applied evolution contexts - where reductions in fitness are common. Toward advancing a more balanced study of evolution, here we present a conceptual framework describing causes of maladaptation. As the introductory article for a Special Feature on maladaptation, we also summarize the studies in this Issue, highlighting the causes of maladaptation in each study. We hope that our framework and the papers in this Special Issue will help catalyze the study of maladaptation in applied evolution, supporting greater understanding of evolutionary dynamics in our rapidly changing world.

Trophic structure modulates community rescue following acidification.

Community rescue occurs when a community that experiences lethal stress persists only through the spread of rare types, either genotypes or species, resistant to the stress. Rescue interacts with trophic structure because physical stress experienced by a focal assemblage within the community may also be experienced by its predators and prey. In general, trophic structure will facilitate rescue only when a stress has a less severe effect on a focal assemblage than on its predators. In other circumstances, when stress affects prey or has only a weak effect on predators, trophic structure is likely to hamper rescue. We exposed a community of phytoplankton and zooplankton derived from a natural lake to acidification in outdoor mesocosms large enough to support trophically complex communities. Rescue of the phytoplankton from severe acidification was facilitated by prior exposure to sublethal stress, confirming previous results from microcosm experiments. Even communities that have previously been less highly stressed were eventually rescued, however, because their zooplankton predators were more sensitive to acidification and became extinct. Our experiment shows how community rescue following severe stress is modulated by the differential effect of the stress relative to trophic level.

Microparasite dispersal in metapopulations: a boon or bane to the host population?

Although connectivity can promote host species persistence in a metapopulation, dispersal may also enable disease transmission, an effect further complicated by the impact that parasite distribution may have on host-parasite population dynamics. We investigated the effects of connectivity and initial parasite distribution (clustered or dispersed) on microparasite-host dynamics in experimental metapopulations, using guppies and Gyrodactylus turnbulli We created metapopulations of guppies divided into four subpopulations and introduced either a low level of parasites to all subpopulations (dispersed) or a high level of parasites to one subpopulation (clustered). Controlled migration among subpopulations occurred every 10 days. In additional trials, we introduced low or high levels of parasites to isolated populations. Parasites persisted longer in metapopulations than in isolated populations. Mortality was lowest in isolated populations with low-level introductions. The interaction of connectivity and initial parasite distribution influenced parasite abundance. With low-level introductions, connectivity helped the parasite persist longer but had little effect on the hosts. With high levels, connectivity also benefited the hosts, lowering parasite burdens. These findings have implications for disease management and species conservation.

Stabilizing mechanisms in a food web with an introduced omnivore.

Intraguild predation (IGP) is an omnivorous food web configuration in which the top predator consumes both a competitor (consumer) and a second prey that it shares with the competitor. This omnivorous configuration occurs frequently in food webs, but theory suggests that it is unstable unless stabilizing mechanisms exist that can decrease the strength of the omnivore and consumer interaction. Although these mechanisms have been documented in native food webs, little is known about whether they operate in the context of an introduced species. Here, we study a marine mussel aquaculture system where the introduction of omnivorous mussels should generate an unstable food web that favors the extinction of the consumer, yet it persists. Using field and laboratory approaches, we searched for stabilizing mechanisms that could reduce interaction strengths in the food web. While field zooplankton counts suggested that mussels influence the composition and abundance of copepods, stable isotope results indicated that life-history omnivory and cannibalism facilitated the availability of prey refugia, and reduced competition and the interaction strength between the mussel omnivore and zooplankton consumers. In laboratory experiments, however, we found no evidence of adaptive feeding which could weaken predator-consumer interactions. Our food web study suggests that the impact of an introduced omnivore may not only depend on its interaction with native species but also on the availability of stabilizing mechanisms that alter the strength of those interactions.

Communities that thrive in extreme conditions captured from a freshwater lake.

Organisms that can grow in extreme conditions would be expected to be confined to extreme environments. However, we were able to capture highly productive communities of algae and bacteria capable of growing in acidic (pH 2), basic (pH 12) and saline (40 ppt) conditions from an ordinary freshwater lake. Microbial communities may thus include taxa that are highly productive in conditions that are far outside the range of conditions experienced in their host ecosystem. The organisms we captured were not obligate extremophiles, but were capable of growing in both extreme and benign conditions. The ability to grow in extreme conditions may thus be a common functional attribute in microbial communities.

Potential for Local Fertilization: A Benthocosm Test of Long-Term and Short-Term Effects of Mussel Excretion on the Plankton.

Mussel aquaculture has expanded worldwide and it is important to assess its impact on the water column and the planktonic food web to determine the sustainability of farming practices. Mussel farming may affect the planktonic food web indirectly by excreting bioavailable nutrients in the water column (a short-term effect) or by increasing nutrient effluxes from biodeposit-enriched sediments (a long-term effect). We tested both of these indirect effects in a lagoon by using plankton-enclosing benthocosms that were placed on the bottom of a shallow lagoon either inside of a mussel farm or at reference sites with no history of aquaculture. At each site, half of the benthocosms were enriched with seawater that had held mussels (excretion treatment), the other half received non-enriched seawater as a control treatment. We monitored nutrients ([PO43-] and [NH4+]), dissolved oxygen and plankton components (bacteria, the phytoplankton and the zooplankton) over 5 days. We found a significant relationship between long-term accumulation of mussel biodeposits in sediments, water-column nutrient concentrations and plankton growth. Effects of mussel excretion were not detected, too weak to be significant given the spatial and temporal variability observed in the lagoon. Effects of mussels on the water column are thus likely to be coupled to benthic processes in such semi-enclosed water bodies.

Predicting the outcome of competition when fitness inequality is variable.

Traditional niche theory predicts that when species compete for one limiting resource in simple ecological settings the more fit competitor should exclude the less fit competitor. Since the advent of neutral theory ecologists have increasingly become interested both in how the magnitude of fitness inequality between competitors and stochasticity may affect this prediction. We used numerical simulations to investigate the outcome of two-species resource competition along gradients of fitness inequality (inequality in R*) and initial population size in the presence of demographic stochasticity. We found that the deterministic prediction of more fit competitors excluding less fit competitors was often unobserved when fitness inequalities were low or stochasticity was strong, and unexpected outcomes such as dominance by the less fit competitor, long-term co-persistence of both competitors or the extinction of both competitors could be common. By examining the interaction between fitness inequality and stochasticity our results mark the range of parameter space in which the predictions of niche theory break down most severely, and suggest that questions about whether competitive dynamics are driven by neutral or niche processes may be locally contingent.

Demasculinization of male guppies increases resistance to a common and harmful ectoparasite.

Parasites are detrimental to host fitness and therefore should strongly select for host defence mechanisms. Yet, hosts vary considerably in their observed parasite loads. One notable source of inter-individual variation in parasitism is host sex. Such variation could be caused by the immunomodulatory effects of gonadal steroids. Here we assess the influence of gonadal steroids on the ability of guppies (Poecilia reticulata) to defend themselves against a common and deleterious parasite (Gyrodactylus turnbulli). Adult male guppies underwent 31 days of artificial demasculinization with the androgen receptor-antagonist flutamide, or feminization with a combination of flutamide and the synthetic oestrogen 17 ß-estradiol, and their parasite loads were compared over time to untreated males and females. Both demasculinized and feminized male guppies had lower G. turnbulli loads than the untreated males and females, but this effect appeared to be mainly the result of demasculinization, with feminization having no additional measurable effect. Furthermore, demasculinized males, feminized males and untreated females all suffered lower Gyrodactylus-induced mortality than untreated males. Together, these results suggest that androgens reduce the ability of guppies to control parasite loads, and modulate resistance to and survival from infection. We discuss the relevance of these findings for understanding constraints on the evolution of resistance in guppies and other vertebrates.

Testing for local host-parasite adaptation: an experiment with Gyrodactylus ectoparasites and guppy hosts.

Hosts and parasites are in a perpetual co-evolutionary "arms race". Due to their short generation time and large reproductive output, parasites are commonly believed to be ahead in this race, although increasing evidence exists that parasites are not always ahead in the arms race - in part owing to evolutionary lineage and recent ecological history. We assess local adaptation of hosts and parasites, and determine whether adaptation was influenced by ecological or evolutionary history, using full reciprocal cross-infections of four Gyrodactylus ectoparasite populations and their four guppy (Poecilia reticulata) host populations in Trinidad. To consider effects of evolutionary lineage and recent ecology, these four populations were collected from two different river drainages (Marianne and Aripo) and two different predation environments (high and low). The highest infection levels were obtained when parasites from the Aripo lineage infected guppies from the Marianne lineage, indicating a higher infectivity, virulence and/or reproductive success of the Aripo parasites. Aripo lineage guppies were also better able to limit Gyrodactylus population growth than guppies from the Marianne River, indicating their strong "resistance" to Gyrodactylus regardless of the source of the parasite. Predation environment had no detectable influence on host-parasite population dynamics of sympatric or allopatric combinations. The much stronger effect of evolutionary lineage (i.e., river) than recent ecological history (i.e., predation) emphasises its importance in driving co-evolutionary dynamics, and should be explored further in future studies on local host-parasite adaptation.

CO2 alters community composition and response to nutrient enrichment of freshwater phytoplankton.

Nutrients can limit the productivity of ecosystems and control the composition of the communities of organisms that inhabit them. Humans are causing atmospheric CO2 concentrations to reach levels higher than those of the past millions of years while at the same time propagating eutrophication through the addition of nutrients to lakes and rivers. We studied the effect of elevated CO2 concentrations, nutrient addition and their interaction in a series of freshwater mesocosm experiments using a factorial design. Our results highlight the important role of CO2 in shaping phytoplankton communities and their response to nutrient addition. We found that CO2 greatly magnified the increase in phytoplankton growth caused by the increased availability of nutrients. Elevated CO2 also caused changes in phytoplankton community composition. As predicted from physiology and laboratory experiments, the taxonomic group that was most limited by current day CO2 concentrations, chlorophytes, increased in relative frequency at elevated CO2. This predictable change in community composition with changes in CO2 is not altered by changes in the availability of other nutrients.