Short and long-term phytoremediation capacity of aquatic plants in Cu-polluted environments.

Freshwater ecosystems face numerous threats from human populations, including heavy metal contamination. Phytoremediation, the use of plants to remediate contaminated soils and sediments, is an effective and low-cost means of removing chemical contaminants, including heavy metals, from polluted environments. However, key questions remain unanswered in the application of this technology in aquatic environments, such as the long-term fate of pollutants following plant uptake. In this study, using two common wetland plant species (duckweed and tape grass), we first examined the capacity of plants to remove copper (Cu) from polluted water. Next, we evaluated the leaching potential of plant tissues following decomposition and how it is affected by a simulated freeze-thaw cycle. Using phytoremediated water and leachates from senesced plants we assessed phytoremediation success and Cu leaching potential by conducting standard toxicity assays using pond snails (Physa acuta), a species with known Cu sensitivity. We found that duckweed outperformed tape grass as a phytoremediator at low Cu concentrations. In addition, for plants grown in low concentrations of Cu, leaching from decaying plant material did not negatively impact snail survival, while at high concentrations of Cu, leaching did result in toxicity. Lastly, we found that a simulated freeze-thaw cycle increased the release of Cu from plant tissue in the presence of high Cu concentrations only, resulting in reduced snail survival. Our results indicate that in moderately Cu-polluted environments, some aquatic plants can remove contaminants without a long-term risk of leaching. In contrast, phytoremediation in highly polluted environments will likely require removal of plant tissue to prevent leaching of previously accumulated metals. Land managers must not only consider plant species and degree of contamination, but also geographic location, as freeze-thaw cycles may enhance plant decomposition and increase the likelihood of contaminant leaching following phytoremediation efforts in aquatic ecosystems.

Local adaptation of the MHC class IIß gene in populations of wood frogs (Lithobates sylvaticus) correlates with proximity to agriculture.

Major histocompatibility complex (MHC) genes code for membrane-embedded proteins that are involved in parasite/pathogen recognition. The link between the MHC and immunity makes these genes important genetic markers to evaluate in systems where infectious disease is associated with population declines. As human impacts on wildlife populations continue to increase, it is also essential to evaluate the role of MHC and immunity in the context of anthropogenic change. Amphibians are an ideal model to test the role of the MHC in infectious disease resistance, as parasites and anthropogenic disturbances currently threaten populations worldwide. We characterized the diversity of MHC class IIß peptide binding region alleles, 13 microsatellite loci, and population-level trematode resistance in 14 populations of wood frogs (Lithobates sylvaticus) in northwestern Pennsylvania with varying geographic distances to agriculture. To assess local adaptation in the MHC IIß, we compared genetic differentiation of MHC IIß and microsatellite markers (FST). We also tested for an effect of isolation by distance on genetic differentiation of MHC IIß and microsatellite markers. In addition, we evaluated whether population-level MHC IIß diversity and common allele frequencies correlate with distance to agriculture and trematode resistance. We found no evidence for genetic structure based on microsatellite analysis nor an effect of isolation by distance on neutral and immunogenetic markers. However, we did detect structure based on the MHC IIß locus, suggesting that it is under local selection. The MHC IIß allele Lisy-DAB*1 was more common in populations living near agricultural sites. Populations with higher MHC IIß diversity showed increased resistance to trematodes. Our results suggest that wood frog populations experience immunogenetic differences at a small scale. In addition, agriculture may disturb natural associations between hosts and parasites through its influence on immunocompetence, underscoring the importance of examining the effects of environmental context on host-parasite interactions.

The effects of different cold-temperature regimes on development, growth, and susceptibility to an abiotic and biotic stressor.

Global climate change is expected to both increase average temperatures as well as temperature variability.Increased average temperatures have led to earlier breeding in many spring-breeding organisms. However, individuals breeding earlier will also face increased temperature fluctuations, including exposure to potentially harmful cold-temperature regimes during early developmental stages.Using a model spring-breeding amphibian, we investigated how embryonic exposure to different cold-temperature regimes (control, cold-pulse, and cold-press) affected (a) compensatory larval development and growth, (b) larval susceptibility to a common contaminant, and (c) larval susceptibility to parasites.We found: (a) no evidence of compensatory development or growth, (b) larvae exposed to the cold-press treatment were more susceptible to NaCl at 4-days post-hatching but recovered by 17-days post-hatching, and (c) larvae exposed to both cold treatments were less susceptible to parasites.These results demonstrate that variation in cold-temperature regimes can lead to unique direct and indirect effects on larval growth, development, and response to stressors. This underscores the importance of considering cold-temperature variability and not just increased average temperatures when examining the impacts of climate disruption.

Inducible pesticide tolerance in Daphnia pulex influenced by resource availability.

Pesticides are a ubiquitous contaminant in aquatic ecosystems. Despite the relative sensitivity of aquatic species to pesticides, growing evidence suggests that populations can respond to pesticides by evolving higher baseline tolerance or inducing a higher tolerance via phenotypic plasticity. While both mechanisms can allow organisms to persist when faced with pesticides, resource allocation theory suggests that tolerance may be related to resource acquisition by the organism. Using Daphnia pulex, we investigated how algal resource availability influenced the baseline and inducible tolerance of D. pulex to a carbamate insecticide, carbaryl. Individuals reared in high resource environments had a higher baseline carbaryl tolerance compared to those reared in low resource environments. However, D. pulex from low resource treatments exposed to sublethal concentrations of carbaryl early in development induced increased tolerance to a lethal concentration of carbaryl later in life. Only individuals reared in the low resource environment induced carbaryl tolerance. Collectively, this highlights the importance of considering resource availability in our understanding of pesticide tolerance.

Patterns of among- and within-species variation in heterospecific pollen receipt: The importance of ecological generalization.

PREMISE OF THE STUDY: Coflowering plants are at risk for receiving pollen from heterospecifics as well as conspecifics, yet evidence shows wide variation in the degree that heterospecific pollen transfer occurs. Evaluation of patterns and correlates of among- and within-species variation in heterospecific pollen (HP) receipt is key to understanding its importance for floral evolution and species coexistence; however, the rarity of deeply sampled multispecies comparisons has precluded such an evaluation. METHODS: We evaluated patterns of among- and within-species variation in HP load size and diversity in 19 species across three distinct plant communities. We assessed the importance of phenotypic specialization (floral phenotype), ecological specialization (contemporary visitor assemblage), and conspecific flower density as determinants of among-species variation. We present hypotheses for different accrual patterns of HP within species based on the evenness and quality of floral visitors and evaluated these by characterizing the relationship between conspecific pollen (CP) and HP receipt. KEY RESULTS: We found that within-species variation in HP receipt was greater than among-species and among-communities variation. Among species, ecological generalization emerged as the strongest driver of variation in HP receipt irrespective of phenotypic specialization. Within-species variation in HP load size and diversity was predicted most often from two CP-HP relationships (linear or exponentially decreasing), suggesting that two distinct types of plant-pollinator interactions prevail. CONCLUSIONS: Our results give important insights into the potential drivers of among- and within-species variation in HP receipt. They also highlight the value of explorations of patterns at the intraspecific level, which can ultimately shed light on plant-pollinator-mediated selection in diverse plant communities.

A first test of elemental allelopathy via heterospecific pollen receipt.

PREMISE OF THE STUDY: Coflowering plants often share pollinators and may receive mixed species pollen loads. Although detrimental effects of heterospecific pollen receipt have been documented, trait-based modifiers of interactions on the stigma remain largely unknown. Chemicals that mediate interactions between sporophytes could also influence pollen-pollen or pollen-style interactions. We test for the first time whether nickel (Ni) accumulation in pollen can lead to "elemental allelopathy" and intensify the fitness consequences of heterospecific pollen receipt. METHODS: We grew Ni-hyperaccumulator Streptanthus polygaloides in soils augmented with three concentrations of Ni, measured pollen Ni concentration, and hand-pollinated non-Ni hyperaccumulator Mimulus guttatus. We assayed pollen germination, tube growth and seeds of M. guttatus after pure and mixed species pollinations. KEY RESULTS: Streptanthus polygaloides pollen accumulated Ni in proportion to soil availability and at levels significantly greater than M. guttatus pollen. Although receipt of S. polygaloides pollen increased M. guttatus pollen germination, it decreased the proportion of pollen tubes reaching the ovary and seed number. Increased Ni in pollen, however, did not significantly intensify the effect of S. polygaloides pollen receipt on M. guttatus seed production. CONCLUSIONS: Different levels of Ni in the pollen of S. polygaloides achieved in the greenhouse did not significantly reduce the fitness of M. guttatus. Stigma tolerance to Ni may also have contributed to the lack of response to increased Ni in heterospecific pollen. This study paves the way for additional tests in other metal hyperaccumulators and recipients, and to identify mechanisms of interactions on the stigma.

Herbivory and relative growth rates of Pieris rapae are correlated with host constitutive salicylic acid and flowering time.

Treatment of plants with exogenous salicylic acid (SA) improves resistance to many bacterial pathogens, but can suppress resistance to insect herbivores. While plants vary naturally in constitutive SA, whether such differences are predictive of resistance to insect herbivores has not been studied previously. We examined the possible role of this endogenous SA in structuring the interactions between the cabbage white butterfly, Pieris rapae, and ten hosts in the mustard family (Brassicaceae). Because P. rapae has multiple generations that utilize different hosts across the year, we included five spring-flowering mustards and five summer-flowering mustards that co-occur in ruderal habitats in upstate New York. Under common garden conditions, the spring flowering mustards (Capsella bursa-pastoris, Draba verna, Cardamine impatiens, Barbarea vulgaris, and Arabidopsis thaliana) were significantly more resistant to P. rapae, supporting 42 % less herbivory (P = 0.015) and 64 % lower relative growth rates (P = 0.007), relative to the summer flowering mustards (Sisymbrium altissimum, Brassica nigra, Sinapis arvense, Lepidium campestre, and Arabis canadensis). Leaf total constitutive SA explained significant variation in larval herbivory (R (2) = 75.3 %, P = 0.007) and relative growth rates (R (2) = 59.4 %, P = 0.043). The three species with the lowest levels of constitutive SA (Capsella bursa-pastoris, Draba verna, and Cardamine impatiens) were the most resistant to larvae. Barbarea vulgaris and Arabis canadensis were notable exceptions, exhibiting high SA concentrations and intermediate resistance to P. rapae. These results suggest a curvilinear relationship between leaf constitutive SA and the herbivory by P. rapae, and they provide some insight into the ecology and possible management of this economically important crop pest.

Effects of floral metal accumulation on floral visitor communities: introducing the elemental filter hypothesis.

UNLABELLED: • PREMISE OF THE STUDY: For plant species that occur in heavy-metal-rich soil, floral metal accumulation may produce an "elemental filter" that reduces pollinator visitation rate and species richness and changes pollinator species composition relative to closely related species growing on normal soils. Consequently, metal hyperaccumulation may contribute to pollinator-mediated reproductive isolation between closely related plant species that differ in metal accumulation.• METHODS: To test these ideas, we characterized plant-pollinator interactions in a sympatric pair of species that differ in metal accumulation (Streptanthus polygaloides, a nickel (Ni) hyperaccumulator, and S. tortuosus, a nonaccumulator). To test the elemental filter hypothesis, we presented arrays of S. polygaloides that were grown in either Ni-treated or control soils to insects at both S. polygaloides and S. tortuosus sites and recorded visitation.• KEY RESULTS: Naturally occurring S. polygaloides hyperaccumulated Ni in anthers and accumulated Ni in nectar, while S. tortuosus did not. Floral visitation rates in natural populations were higher to S. tortuosus than S. polygaloides. In addition, while floral visitor richness was similar, few pollinator taxa were shared between the two plant species. Nickel-treatment of S. polygaloides reduced visits by bees, but only for arrays presented at S. tortuosus sites.• CONCLUSIONS: We show that the Ni hyperaccumulator S. polygaloides hosts a distinct floral visitor community, indicating that metal accumulation creates a filter for pollinators, similar to that documented for herbivores. Our study highlights a novel mechanism by which the abiotic environment can alter plant-pollinator interactions, and consequently plant reproduction and speciation.

Nickel accumulation in leaves, floral organs and rewards varies by serpentine soil affinity.

Serpentine soils are edaphically stressful environments that host many endemic plant species. In particular, serpentine soils are high in several heavy metals (e.g. nickel, cobalt and chromium) and these high heavy metal concentrations are thought, in part, to lead to varying levels of plant adaptation and soil affinities (i.e. endemic vs. non-endemic plant species). It is unclear, however, whether serpentine endemics vs. non-endemics differ with respect to heavy metal uptake into either vegetative or reproductive organs. Here, we use nickel as a model to determine whether plant heavy metal uptake varies with the level of endemism in several non-hyperaccumulating species. Under controlled greenhouse conditions, we grew seven plant species from the Brassicaceae family that vary in their degrees of affinity to serpentine soil from low (indifferent) to medium (indicator) and high (endemic) in soil that was nickel supplemented or not. We quantified nickel concentrations in leaves, pistils, anthers, pollen and nectar. While nickel concentrations did not vary across organs or affinities when grown in control soils, under conditions of nickel supplementation endemic species had the lowest tissue concentrations of nickel, particularly when considering leaves and pistils, compared with indifferent/indicator species. Species indifferent to serpentines incorporated higher concentrations of nickel into reproductive organs relative to leaves, but this was not the case for indicator species and endemics where nickel concentration was similar in these organs. Our findings suggest that endemic species possess the ability to limit nickel uptake into above-ground tissues, particularly in reproductive organs where it may interfere with survival and reproduction. Indifferent species accumulated significantly more nickel into reproductive organs compared with leaves, which may limit their reproductive potential relative to endemic species when growing on serpentine soils. Additional work determining the fitness consequences of these differences will further our understanding of edaphic endemism.

Nickel accumulation by Streptanthus polygaloides (Brassicaceae) reduces floral visitation rate.

Hyperaccumulation is the phenomenon whereby plants take up and sequester in high concentrations elements that generally are excluded from above-ground tissues. It largely is unknown whether the metals taken up by these plants are transferred to floral rewards (i.e., nectar and pollen) and, if so, whether floral visitation is affected. We grew Streptanthus polygaloides, a nickel (Ni) hyperaccumulator, in short-term Ni supplemented soils and control soils to determine whether Ni is accumulated in floral rewards and whether floral visitation is affected by growth in Ni-rich soils. We found that while supplementation of soils with Ni did not alter floral morphology or reward quantity (i.e., anther size or nectar volume), Ni did accumulate in the nectar and pollen-filled anthers-providing the first demonstration that Ni is accumulated in pollinator rewards. Further, S. polygaloides grown in Ni-supplemented soils received fewer visits per flower per hour from both bees and flies (both naïve to Ni-rich floral resources in the study area) relative to plants grown in control soils, although the probability a plant was visited initially was unaffected by Ni treatment. Our findings show that while Ni-rich floral rewards decrease floral visitation, floral visitors are not completely deterred, so some floral visitors may collect and ingest potentially toxic resources from metal-hyperaccumulating plants. In addition to broadening our understanding of the effects of metal accumulation on ecological interactions in natural populations, these results have implications for the use of insect-pollinated plants in phytoremediation.

Among-species differences in pollen quality and quantity limitation: implications for endemics in biodiverse hotspots.

BACKGROUND AND AIMS: Insufficient pollination is a function of quantity and quality of pollen receipt, and the relative contribution of each to pollen limitation may vary with intrinsic plant traits and extrinsic ecological properties. Community-level studies are essential to evaluate variation across species in quality limitation under common ecological conditions. This study examined whether endemic species are more limited by pollen quantity or quality than non-endemic co-flowering species in three endemic-rich plant communities located in biodiversity hotspots of different continents (Andalusia, California and Yucatan). METHODS: Natural variations in pollen receipt and pollen tube formation were analysed for 20 insect-pollinated plants. Endemic and non-endemic species that co-flowered were paired in order to estimate and compare the quantity and quality components of pre-zygotic pollination success, obtained through piecewise regression analysis of the relationship between pollen grains and pollen tubes of naturally pollinated wilted flowers. KEY RESULTS: Pollen tubes did not frequently exceed the number of ovules per flower. Only the combination of abundant and good quality pollen and a low number of ovules per flower conferred relief from pre-zygotic pollen limitation in the three stochastic pollination environments studied. Quality of pollen receipt was found to be as variable as quantity among study species. The relative pollination success of endemic and non-endemic species, and its quantity and quality components, was community dependent. CONCLUSIONS: Assessing both quality and quantity of pollen receipt is key to determining the ovule fertilization potential of both endemic and widespread plants in biodiverse hotspot regions. Large natural variation among flowers of the same species in the two components and pollen tube formation deserves further analysis in order to estimate the environmental, phenotypic and intraindividual sources of variation that may affect how plants evolve to overcome this limitation in different communities worldwide.

Edaphic factors and plant-insect interactions: direct and indirect effects of serpentine soil on florivores and pollinators.

Edaphic factors can lead to differences in plant morphology and tissue chemistry. However, whether these differences result in altered plant-insect interactions for soil-generalist plants is less understood. We present evidence that soil chemistry can alter plant-insect interactions both directly, through chemical composition of plant tissue, and indirectly, through plant morphology, for serpentine-tolerant Mimulus guttatus (Phrymaceae). First, we scored floral display (corolla width, number of open flowers per inflorescence, and inflorescence height), flower chemistry, pollinator visitation and florivory of M. guttatus growing on natural serpentine and non-serpentine soil over 2 years. Second, we conducted a common garden reciprocal soil transplant experiment to isolate the effect of serpentine soil on floral display traits and flower chemistry. And last, we observed arrays of field-collected inflorescences and potted plants to determine the effect of soil environment in the field on pollinator visitation and florivore damage, respectively. For both natural and experimental plants, serpentine soil caused reductions in floral display and directly altered flower tissue chemistry. Plants in natural serpentine populations received fewer pollinator visits and less damage by florivores relative to non-serpentine plants. In experimental arrays, soil environment did not influence pollinator visitation (though larger flowers were visited more frequently), but did alter florivore damage, with serpentine-grown plants receiving less damage. Our results demonstrate that the soil environment can directly and indirectly affect plant-mutualist and plant-antagonist interactions of serpentine-tolerant plants by altering flower chemistry and floral display.

The effects of aluminum and nickel in nectar on the foraging behavior of bumblebees.

Metals in soil are known to negatively affect the health of many groups of organisms, but it is unclear whether they can affect plant-pollinator interactions, and whether pollinators that visit plants growing on contaminated soils are at risk of ingesting potentially toxic resources. We address whether the presence of metals in nectar alters foraging behavior by bumblebees by manipulating nectar with one of two common soil contaminants (Al or Ni) in flowers of Impatiens capensis (Balsaminaceae). While the presence of Al in nectar did not influence foraging patterns by bumblebees, flowers containing Ni nectar solutions were visited for shorter time periods relative to controls, and discouraged bees from visiting nearby Ni-contaminated flowers. However, because bumblebees still visited these flowers, they likely ingested a potentially toxic resource. Our findings suggest that soil metals could cascade to negatively affect pollinators in metal contaminated environments.

Influence of fire on a rare serpentine plant assemblage: a 5-year study of Darlingtonia fens.

PREMISE OF THE STUDY: Serpentine soils have attracted the attention of evolutionary biologists for decades because of their high number of rare and endemic taxa, though less is known about the ecological factors that govern the diversity and composition of serpentine communities. Theory suggests that vegetation on these low-productivity soils will be relatively resilient to fire, the most common natural disturbance in serpentine systems. METHODS: We studied the recovery of vegetation in Darlingtonia fens, a unique habitat dominated by herbaceous perennials, from a major fire that burned ∼202,000 ha in California and Oregon's Klamath Mountains in 2002. We established permanent plots in eight unburned and eight burned fens in 2003 and recorded percent cover of vascular plant species. We re-sampled plots each year through 2007. KEY RESULTS: Burned fens had less plant cover than unburned fens for 2 yr after the fire. Average species density was ∼10% lower in burned fens 1 yr after the fire but ∼4-8% higher for the next 4 yr. Burned fens exhibited greater evenness but not until 4 yr after the fire. Differences in community composition were detected between the two fen types, but species ranks were similar, and species neither were added to nor removed from the burned assemblages. CONCLUSIONS: Burning of Darlingtonia fens has detectable, albeit modest, effects on serpentine communities. Because fens have little or no canopy cover, fire has little influence on light availability in this system. This relatively small resource change, combined with high soil moisture and well-developed underground organs of fen plants, produces a highly resilient assemblage.