Direct and indirect interactions for coexistence in a species-defined microcosm

Mechanisms for coexistence among micro-organisms were studied by using a species-defined microcosm, consisting of the bacterium Escherichia coli, the ciliate Tetrahymena thermophila and the alga Euglena gracilis. These organisms were chosen as representative of ecological functional groups i.e. decomposer, consumer and producer, respectively. Direct and indirect interactions among these organisms were evaluated by comparisons of their population dynamics in culture with different combinations of the three species. There was an E. coli cell density dependent predator–prey interaction between T. thermophila and E. coli which was only established when there were more than 10⁶ cells ml^–1 of E. coli. Indirect interactions were evaluated from the cultivation of each organism in media containing metabolites of the others. Metabolites from each population strongly accelerated the growth of their own populations and those of the others except for the self-toxicity effect of E. coli metabolites. These observations suggested that not only the cell–cell contact of direct interactions, but also metabolite-mediated indirect interactions supported the maintenance of the populations of each micro-organism and their coexistence. In natural ecosystems, there are many interactions and it is difficult to evaluate all those regulating community dynamics. The gnotobiotic microcosm used in this study was shown to be suitable for examining the specific, species–species microbial interactions.     

Carbon dioxide and methane fluxes in boreal peatland microcosms with different vegetation cover—effects of ozone or ultraviolet-B exposure

O₃ concentrations in the troposphere are rising and those in the stratosphere decreasing, the latter resulting in higher fluxes of solar ultraviolet-B (UV-B) radiation to the earth's surface. We assessed whether the fluxes of CO₂ and CH₄ are altered by enhanced UV-B radiation or elevated tropospheric O₃ concentrations in boreal peatland microcosms (core depth 40 cm, diameter 10.5 cm) with different vegetation cover. At the end of the UV-B experiment which lasted for a growing season, net CO₂ exchange (NEE) and dark ecosystem respiration (R _TOT) were sevenfold higher, and CH₄ efflux 12-fold higher, in microcosms with intact vegetation dominated by Eriophorum vaginatum L. and Sphagnum spp., compared to microcosms from which we removed E. vaginatum. Vegetation treatment had minor effects on CH₄ production and consumption potentials in the peat, suggesting that the large difference in CH₄ efflux is mainly due to efficient CH₄ transport via the aerenchyma of E. vaginatum. Ambient UV-B supplemented with 30% and elevated O₃ concentrations (100 and 200 ppb, for 7 weeks) significantly increased R _TOT in both vegetation treatments. Elevated O₃ concentrations reduced NEE over time, while UV-B had no clear effects on the fluxes of CO₂ or CH₄ in the cloudy summer of the study. Field experiments are needed to assess the significance of increasing UV-B radiation and elevated tropospheric O₃ concentration on peatland gas exchange in the long-term.     

Water-soluble vitamins in cells and spent culture supernatants of Poteriochromonas stipitata,Euglena gracilis,and Tetrahymena thermophila

Vitamins B₆ and B₁₂, biotin, folates, riboflavin, nicotinate, pantothenate, biopterin, and vitamin C (l-ascorbate) were assayed in Poteriochromonas stipitata, Euglena gracilis, and Tetrahymena thermophila cells grown in defined media and in spent culture supernatants. P. stipitata and E. gracilis synthesized, stored and excreted folates (mainly as 5-methyltetrahydrofolate), B₆, riboflavin, pantothenate, nicotinate, biopterin, and ascorbate. E. gracilis synthesized and stored biotin. T. thermophila did not synthesize the above vitamins except for B₁₂, biopterin, and ascorbate; it excreted biopterin and stored B₁₂ and ascorbate. Thiamin was left of consideration because all 3 organisms are thiamin auxotrophs. Possible ecological implications of these findings are considered.     

The response of mycorrhizal colonization to elevated CO2 and climate change in Pascopyrum smithii and Bouteloua gracilis

Large intact soil cores of nearly pure stands of Pascopyrum smithii (western wheatgrass, C₃) and Bouteloua gracilis (blue grama, C₄) were extracted from the Central Plains Experimental Range in northeastern Colorado, USA and transferred to controlled environment chambers. Cores were exposed to a variety of water, temperature and CO₂ regimes for a total of four annual growth cycles. Root subsamples were harvested after the completion of the second and fourth growth cycles at a time corresponding to late winter, and were examined microscopically for the presence of mycorrhizae. After two growth cycles in the growth chambers, 54% of the root length was colonized in P. smithii, compared to 35% in blue grama. Field control plants had significantly lower colonization. Elevation of CO₂ increased mycorrhizal colonization in B. gracilis by 46% but had no effect in P. smithii. Temperatures 4° C higher than normal decreased colonization in P. smithii by 15%. Increased annual precipitation decreased colonization in both species. Simulated climate change conditions of elevated CO₂, elevated temperature and lowered precipitation decreased colonization in P. smithii but had less effect on B. gracilis. After four growth cycles in P. smithii, trends of treatments remained similar, but overall colonization rate decreased.     

Nitrogen and Carbon Concentrations, and Stable Isotope Ratios in Mediterranean Shrubs Growing in the Proximity of a CO2 spring

Seasonal changes in foliage nitrogen (N) and carbon (C) concentrations and δ¹⁵N and δ¹³C ratios were monitored during a year in Erica arborea, Myrtus communis and Juniperus communis co-occurring at a natural CO₂ spring (elevated [CO₂], about 700 μmol mol⁻¹) and at a nearby control site (ambient [CO₂], 360 μmol mol⁻¹) in a Mediterranean environment. Leaf N concentration was lower in elevated [CO₂] than in ambient [CO₂] for M. communis, higher for J. communis, and dependent on the season for E. arborea. Leaf C concentration was negatively affected by atmospheric CO₂ enrichment, regardless of the species. C/N ratio varied concomitantly to N. Leaves in elevated [CO₂] showed lower δ¹³C, and therefore likely lower water use efficiencies than leaves at the control site, regardless of the species, suggesting substantial photosynthetic acclimation under long-term CO₂-enriched atmosphere. Leaves of E. arborea showed lower values of δ¹⁵N under elevated [CO₂], but this was not the case of M. communis and J. communis foliage. The use of the resources and leaf chemical composition are affected by elevated [CO₂], but such an effect varies during the year, and is species-dependent. The seasonal dependency and species specificity suggest that plants are able to exploit different available water and N resources within Mediterranean sites. This revised version was published online in July 2006 with corrections to the Cover Date.     

Independent, Interactive, and Species-Specific Responses of Leaf Litter Decomposition to Elevated CO2 and O3 in a Northern Hardwood Forest

The future capacity of forest ecosystems to sequester atmospheric carbon is likely to be influenced by CO₂-mediated shifts in nutrient cycling through changes in litter chemistry, and by interactions with pollutants like O₃. We evaluated the independent and interactive effects of elevated CO₂ (560 μl l⁻¹) and O₃ (55 nl l l⁻¹) on leaf litter decomposition in trembling aspen (Populus tremuloides) and paper birch (Betula papyrifera) at the Aspen free air CO₂ enrichment (FACE) site (Wisconsin, USA). Fumigation treatments consisted of replicated ambient, +CO₂, +O₃, and +CO₂ + O₃ FACE rings. We followed mass loss and litter chemistry over 23 months, using reciprocally transplanted litterbags to separate substrate quality from environment effects. Aspen decayed more slowly than birch across all treatment conditions, and changes in decomposition dynamics of both species were driven by shifts in substrate quality rather than by fumigation environment. Aspen litter produced under elevated CO₂ decayed more slowly than litter produced under ambient CO₂, and this effect was exacerbated by elevated O₃. Similarly, birch litter produced under elevated CO₂ also decayed more slowly than litter produced under ambient CO₂. In contrast to results for aspen, however, elevated O₃ accelerated birch decay under ambient CO₂, but decelerated decay under enriched CO₂. Changes in decomposition rates (k-values) were due to CO₂- and O₃-mediated shifts in litter quality, particularly levels of carbohydrates, nitrogen, and tannins. These results suggest that in early-successional forests of the future, elevated concentrations of CO₂ will likely reduce leaf litter decomposition, although the magnitude of effect will vary among species and in response to interactions with tropospheric O₃.     

Effects of elevated CO2 and O3 on leaf litter phenolics and subsequent performance of litter-feeding soil macrofauna

Two field-growing silver birch (Betula pendula Roth) clones (clone 4 and 80) were exposed to elevated CO₂ and O₃ for three growing seasons (1999–2001). The phenolic compounds of naturally abscised leaf litter were analyzed in order to determine the possible CO₂- and O₃-induced changes in the litter quality. The potential litter-mediated CO₂ and O₃ effects on litter-feeding soil macrofauna (detritivore) performance were assessed in microcosm experiments, i.e., the relative growth rates (RGR) of Lumbricus terrestris and Porcellio scaber, the relative consumption rates (RCR) of P. scaber, and mortality of the test animals were measured. The leaf litter grown under elevated CO₂ had increased concentrations (weight per mass unit) and contents (weight per leaf) of phenolic acids, flavonol glycosides, condensed tannins and total measured phenolics. Elevated O₃ increased the concentrations of 3,4’-dihydroxypropiophenone 3-β-d-glucoside (DHPPG) and flavonoid aglycones but only under ambient CO₂. However, elevated O₃ effects on the content of some low-molecular-weight phenolic (LMWP) compounds (i.e. phenolic acids, DHPPG, flavonoid aglycones) and total LMWP changed over time emphasizing the importance of conducting long-term (>3 years) exposure studies. In general, RGR of young L. terrestris was affected by the litter quality changes induced by elevated CO₂ and O₃, as the animal growth rates were reduced when they were fed with CO₂- and O₃-exposed leaf litter of clone 80 in Experiment 1. P. scaber RCR or RGR responses to CO₂- and O₃-induced changes in litter quality were more variable and inconsistent, and neither were there any litter-mediated CO₂ and O₃ effects on animal mortality in these microcosm experiments. In conclusion, elevated CO₂ has the potential to alter silver birch leaf litter quality, but the possible O₃ effects on phenolic compounds and litter-mediated CO₂ and O₃ effects on detritivores are more difficult to validate.     

Hemiparasite abundance in an alpine treeline ecotone increases in response to atmospheric CO2 enrichment

Populations of the annual hemiparasites Melampyrum pratense L. and Melampyrum sylvaticum L. were studied at the treeline in the Swiss Alps after 3 years of in situ CO₂ enrichment. The total density of Melampyrum doubled to an average of 44 individuals per square meter at elevated CO₂ compared to ambient CO₂. In response to elevated CO₂, the height of the more abundant and more evenly distributed M. pratense increased by 20%, the number of seeds per fruit by 21%, and the total seed dry mass per fruit by 27%, but the individual seed size did not change. These results suggest that rising atmospheric CO₂ may stimulate the reproductive output and increase the abundance of Melampyrum in the alpine treeline ecotone. Because hemiparasites can have important effects on community dynamics and ecosystem processes, notably the N cycle, changing Melampyrum abundance may potentially influence the functioning of alpine ecosystems in a future CO₂-rich atmosphere.     

Does carbon partitioning in ectomycorrhizal pine seedlings under elevated CO2 vary with fungal species?

Enhanced soil respiration in response to elevated atmospheric CO₂ has been demonstrated, and ectomycorrhizal (ECM) fungi are of particular interest since they partition host-derived photoassimilates belowground. Although a strong response of ECM fungi to elevated CO₂ has been shown, little is still known about the functional diversity among species. We studied carbon (C) partitioning in mycorrhizal Scots pine seedlings in response to short-term CO₂ enrichment, using seven ECM species with different ecological strategies. Mycorrhizal associations were synthesised and seedlings grown in large Petri dishes containing peat:vermiculite and nutrient solution for 10–15 weeks, after which half of the microcosms were exposed to elevated CO₂ treatment (710 ppm) for 15 days and the other half were kept in ambient CO₂ treatment. Partitioning of C was quantified by pulse labelling the seedlings with ¹⁴CO₂ and examining the distribution of labelled assimilates in shoot, root and extraradical mycelial compartments by destructive harvest and liquid scintillation counting. Fungal biomass was determined with PLFA analysis. The respiratory loss of ¹⁴CO₂ was on average greater in the elevated CO₂ treatment for most species compared to the ambient CO₂ treatment. More label was retrieved in the shoots in the ambient CO₂ treatment compared to elevated CO₂ (significant for P. involutus and P. fallax). Greater amounts of label were found in the extraradical mycelial compartment in all species (except P. involutus) in elevated CO₂ compared to ambient CO₂ (significant for L. bicolor, P. byssinum, P. fallax and R. roseolus). Fungal biomass production increased significantly with elevated CO₂ for two species (H. velutipes and A. muscaria); three species (P. fallax, P. involutus and R. roseolus) showed a similar but non-significant trend, whereas L. bicolor and P. byssinum produced less biomass in elevated CO₂ compared to ambient CO₂. When ¹⁴C in the mycelial compartment and respiration was expressed per unit fungal PLFA the difference between CO₂ treatments disappeared. We demonstrated that different ECM fungal isolates respond differently in C partitioning in response to CO₂ enrichment. These results suggest that under certain growth conditions, when nutrients are not limiting, ECM fungi respond rapidly to increasing C-availability through changed biomass production and respiration.     

Effect of elevated atmospheric carbon dioxide on the use of foliar application of Bacillus thuringiensis

Toxins from Bacillus thuringiensis have beenused as pest management tools for more than 50 years. The effect of these toxins depends on the quantityof Bacillus thuringiensis (Bt) toxins ingestedby susceptible insects. Food ingestion is affected byCO₂ concentration; plants grown in elevatedCO₂ often have increased carbon/nitrogen ratios(C/N), resulting in greater leaf area consumption. Therefore, we hypothesized that elevated CO₂would improve the efficacy of foliar applications ofB. thuringiensis. Cotton plants were grown ateither ambient (360–380 l/l) or elevated CO₂(900 l/l). Groups of plants in both CO₂treatments were exposed to low (30 mg/kg soil/week) orhigh (130 mg/kg soil/week) nitrogen (N) fertilizationlevels in a split plot design. The resulting plantswere assessed for N and carbon (C) contents. Leafdisks from the same plants were dipped in a Btsolution and then fed to Spodoptera exigua(Hübner), an insect species of considerableeconomic importance. Elevated CO₂ significantlyreduced total N, and increased the C/N. Nitrogenfertilization significantly affected consumption byearly stadia larvae, larval weight gain, and relativegrowth rate (RGR). Interactions between CO₂concentration and N fertilization level significantlyimpacted late stadia larval food consumption, andthrough differential Bt toxin intake, affectedduration of larval stage and mortality to the adultstage. We conclude that the elevated atmosphericCO₂ concentrations expected in the next centurywill interact with commercial fertilization practicesto enhance the efficacy of B. thuringiensisformulations applied topically to crops. Theimplications for improved control are discussed.     

Increases in Desert Shrub Productivity under Elevated Carbon Dioxide Vary with Water Availability

Productivity of aridland plants is predicted to increase substantially with rising atmospheric carbon dioxide (CO₂) concentrations due to enhancement in plant water-use efficiency (WUE). However, to date, there are few detailed analyses of how intact desert vegetation responds to elevated CO₂. From 1998 to 2001, we examined aboveground production, photosynthesis, and water relations within three species exposed to ambient (around 38 Pa) or elevated (55 Pa) CO₂ concentrations at the Nevada Desert Free-Air CO₂ Enrichment (FACE) Facility in southern Nevada, USA. The functional types sampled—evergreen (Larrea tridentata), drought-deciduous (Ambrosia dumosa), and winter-deciduous shrubs (Krameria erecta)—represent potentially different responses to elevated CO₂ in this ecosystem. We found elevated CO₂ significantly increased aboveground production in all three species during an anomalously wet year (1998), with relative production ratios (elevated:ambient CO₂) ranging from 1.59 (Krameria) to 2.31 (Larrea). In three below-average rainfall years (1999–2001), growth was much reduced in all species, with only Ambrosia in 2001 having significantly higher production under elevated CO₂. Integrated photosynthesis (mol CO₂ m⁻² y⁻¹) in the three species was 1.26–2.03-fold higher under elevated CO₂ in the wet year (1998) and 1.32–1.43-fold higher after the third year of reduced rainfall (2001). Instantaneous WUE was also higher in shrubs grown under elevated CO₂. The timing of peak canopy development did not change under elevated CO₂; for example, there was no observed extension of leaf longevity into the dry season in the deciduous species. Similarly, seasonal patterns in CO₂ assimilation did not change, except for Larrea. Therefore, phenological and physiological patterns that characterize Mojave Desert perennials—early-season lags in canopy development behind peak photosynthetic capacity, coupled with reductions in late-season photosynthetic capacity prior to reductions in leaf area—were not significantly affected by elevated CO₂. Together, these findings suggest that elevated CO₂ can enhance the productivity of Mojave Desert shrubs, but this effect is most pronounced during years with abundant rainfall when soil resources are most available.     

CO2 enrichment reduces reproductive dominance in competing stands of Ambrosia artemisiifolia (common ragweed)

Plants growing in dense stands may not equally acquire or utilize extra carbon gained in elevated CO₂. As a result, reproductive differences between dominant and subordinate plants may be altered under rising CO₂ conditions. We hypothesized that elevated CO₂ would enhance the reproductive allocation of shaded, subordinate Ambrosia artemisiifolia L. (Asteraceae) individuals more than that of light-saturated dominants. We grew stands of A. artemisiifolia at either 360 or 720 μL L⁻¹ CO₂ levels and measured the growth and reproductive responses of competing individuals. To test whether elevated CO₂ altered size and reproductive inequalities within stands, we compared stand-level coefficients of variation (CV) in height growth and final shoot, root, and reproductive organ biomasses. Elevated CO₂ enhanced biomass and reduced the CV for all aspects of plant growth, especially reproductive biomass. Allocation to reproduction was higher in the elevated CO₂ than in the ambient treatment, and this difference was more pronounced in small, rather than large plant positive relationships between the CV and total stand productivity declined under elevated CO₂, indicating that growth enhancements to smaller plants diminished the relative biomass advantages of larger plants in increasingly crowded conditions. We conclude that elevated CO₂ stimulates stand-level reproduction while CO₂-induced growth gains of subordinate A. artemisiifolia plants minimize differences in the reproductive output of small and large plants. Thus, more individuals are likely to produce greater amounts of seeds and pollen in future populations of this allergenic weed.     

Gas Exchange Responses to CO2 Concentration Instantaneously Elevated in Flag Leaves of Winter Wheat Cultivars Released in Different Years

Three winter wheat (Triticum aestivum L.) cultivars, representatives of those widely cultivated in Beijing over the past six decades, were grown in the same environmental conditions. Net photosynthetic rate (P _N) per unit leaf area and instantaneous water use efficiency (WUE) of flag leaves increased with elevated CO₂ concentration. With an increase in CO₂ concentration from 360 to 720 µmol mol^–1, P _N and WUE of Jingdong 8 (released in 1990s and having the highest yield) increased by 173 and 81 %, while those of Nongda 139 (released in 1970s) increased by 88 and 66 %, and Yanda 1817 (released in 1945, with lowest yield) by 76 and 65 %. Jingdong 8 had the highest P _N and WUE values under high CO₂ concentration, but Yanda 1817 showed the lowest P _N. Stomatal conductance (g _s) of Nongda 139 and Yanda 1817 declined with increasing CO₂ concentration, but g _s of Jingdong 8 firstly went down and then up as the CO₂ concentration further increased. Intercellular CO₂ concentration (C _i) of Jingdong 8 and Nongda 139 increased when CO₂ concentration elevated, while that of Yanda 139 increased at the first stage and then declined. Jingdong 8 had the lowest C _i of the three wheat cultivars, and Yanda 1817 had the highest C _i value under lower CO₂ concentrations. However, Jingdong 8 had the highest P _N and lowest C _i at the highest CO₂ concentration which indicates that its photosynthetic potential may be high.     

Effects of drainage and temperature on carbon balance of tussock tundra micrososms

We examined the importance of temperature (7°C or 15°C) and soil moisture regime (saturated or field capacity) on the carbon (C) balance of arctic tussock tundra microcosms (intact blocks of soil and vegetation) in growth chambers over an 81-day simulated growing season. We measured gaseous CO₂ exchanges, methane (CH₄) emissions, and dissolved C losses on intact blocks of tussock (Eriophorum vaginatum) and intertussock (moss-dominated). We hypothesized that under increased temperature and/or enhanced drainage, C losses from ecosystem respiration (CO₂ respired by plants and heterotrophs) would exceed gains from gross photosynthesis causing tussock tundra to become a net source of C to the atmosphere. The field capacity moisture regime caused a decrease in net CO₂ storage (NEP) in tussock tundra micrososms. This resulted from a stimulation of ecosystem respiration (probably mostly microbial) with enhanced drainage, rather than a decrease in gross photosynthesis. Elevated temperature alone had no effect on NEP because CO₂ losses from increased ecosystem respiration at elevated temperature were compensated by increased CO₂ uptake (gross photosynthesis). Although CO₂ losses from ecosystem respiration were primarily limited by drainage, CH₄ emissions, in contrast, were dependent on temperature. Furthermore, substantial dissolved C losses, especially organic C, and important microhabitat differences must be considered in estimating C balance for the tussock tundra system. As much as 20% of total C fixed in photosynthesis was lost as dissolved organic C. Tussocks stored 2x more C and emitted 5x more methane than intertussocks. In spite of the limitations of this microcosm experiment, this study has further elucidated the critical role of soil moisture regime and dissolved C losses in regulating net C balance of arctic tussock tundra.     

Leaf and canopy responses to elevated CO2 in a pine forest under free-air CO2 enrichment

Physiological responses to elevated CO₂ at the leaf and canopy-level were studied in an intact pine (Pinus taeda) forest ecosystem exposed to elevated CO₂ using a free-air CO₂ enrichment (FACE) technique. Normalized canopy water-use of trees exposed to elevated CO₂ over an 8-day exposure period was similar to that of trees exposed to current ambient CO₂ under sunny conditions. During a portion of the exposure period when sky conditions were cloudy, CO₂-exposed trees showed minor (7%) but significant reductions in relative sap flux density compared to trees under ambient CO₂ conditions. Short-term (minutes) direct stomatal responses to elevated CO₂ were also relatively weak (5% reduction in stomatal aperture in response to high CO₂ concentrations). We observed no evidence of adjustment in stomatal conductance in foliage grown under elevated CO₂ for nearly 80 days compared to foliage grown under current ambient CO₂, so intrinsic leaf water-use efficiency at elevated CO₂ was enhanced primarily by direct responses of photosynthesis to CO₂. We did not detect statistical differences in parameters from photosynthetic responses to intercellular CO₂ (A _net-C _i curves) for Pinus taeda foliage grown under elevated CO₂ (550 mol mol^–1) for 50–80 days compared to those for foliage grown under current ambient CO₂ from similar-sized reference trees nearby. In both cases, leaf net photosynthetic rate at 550 mol mol^–1 CO₂ was enhanced by approximately 65% compared to the rate at ambient CO₂ (350 mol mol^–1). A similar level of enhancement under elevated CO₂ was observed for daily photosynthesis under field conditions on a sunny day. While enhancement of photosynthesis by elevated CO₂ during the study period appears to be primarily attributable to direct photosynthetic responses to CO₂ in the pine forest, longer-term CO₂ responses and feedbacks remain to be evaluated.     

A consideration of Euglena gracilis W3BUL as a cytoplasmic control for the wild-type phenylalanyl-tRNA synthetase system

The studies described indicate that the UV bleached mutant, Euglena gracilis W₃BUL does not serve as a suitable cytoplasmic control for the phenylalanyl-tRNA synthetase system. Chromatography of wild-type E. gracilis on Sephadex G100 revealed three peaks of activity identified as the chloroplastic, cytoplasmic and mitochondrial enzymes. The chloroplastic activity was greater in log than in stationary phase cells and was the only activity recovered from purified chloroplasts. Cell-free extracts of the achloroplastic mutant, E. gracilis W₃BUL, contained wild-type levels of the cytoplasmic and mitochondrial phenylalanyl-tRNA synthetases. However, no chloroplastic synthetase was detected in the mutant extracts. Anomalies in the aminoacylation behavior of the W₃BUL system were observed which suggest the possibility of a mutation affecting non-chloroplastic tRNAs in this UV-induced mutant. These anomalies significantly reduce the ability of the E. gracilis W₃BUL mutant to serve as a cytoplasmic control in the phenylalanyl-tRNA synthetase system.     

Adaptive characteristics of a ciliateTetrahymena thermophila in endosymbiotic association with a green algaChlorella vulgaris derived in a long-term microcosm culture

We investigated directly an early stage of the development of an endosymbiosis between an alga and a ciliate, beginning from a non-associated stage by conducting a long-term microcosm culture composed of a green alga (Chlorella vulgaris), a bacterium (Escherichia coli) and a ciliate (Tetrahymena thermophila) for three years. During this culture,Chlorella-harboringTetrahymena appeared and increased in frequency. We examined the adaptive characteristics of theT. thermophila in association with theC. vulgaris derived from a well-established microcosm culture maintained for 1164–1400 days, and compared it with the original culture ofT. thermophila. TheT. thermophila with the associated, intracellularC. vulgaris grew at a lower density ofE. coli than the originalT. thermophila with the originalC. vulgaris, and the former survived longer than the latter in the absence ofE. coli. These results suggest that this induced algal-ciliate association confers some fitness advantage at least to the host under the conditions of less or no available food such asE. coli. Although the algal-ciliate association is not completely stable, this quasi-stable association may enable the host to exploit a new niche through the advantages of mixotrophy.     

Inter- and intra-generic differences in growth,reproduction, and fitness of nine herbaceous annual species grown in elevated CO2 environments

In assessing the capacity of plants to adapt to rapidly changing global climate, we must elucidate the impacts of elevated carbon dioxide on reproduction, fitness and evolution. We investigated how elevated CO₂ influenced reproduction and growth of plants exhibiting a range of floral morphologies, the implications of shifts in allocation for fitness in these species, and whether related taxa would show similar patterns of response. Three herbaceous, annual species each of the genera Polygonum, Ipomoea, and Cassia were grown under 350 or 700 ppm CO₂. Vegetative growth and reproductive output were measured non-destructively throughout the full life span, and vegetative biomass was quantified for a subsample of plants in a harvest at first flowering. Viability and germination studies of seed progeny were conducted to characterize fitness precisely. Early vegetative growth was often enhanced in high-CO₂ grown plants of Polygonum and Cassia (but not Ipomoea). However, early vegetative growth was not a strong predictor of subsequent reproduction. Phenology and production of floral buds, flowers, unripe and abscised fruits differed between CO₂ treatments, and genera differed in their reproductive and fitness responses to elevated CO₂. Polygonum and Cassia species showed accelerated, enhanced reproduction, while Ipomoea species generally declined in reproductive output in elevated CO₂. Seed quality and fitness (in terms of viability and percentage germination) were not always directly correlated with quantity produced, indicating that output alone may not reliably indicate fitness or evolutionary potential. Species within genera typically responded more consistently to CO₂ than unrelated species. Cluster analyses were performed separately on suites of vegetative and reproductive characters. Some species assorted within genera when these reproductive responses were considered, but vegetative responses did not reflect taxonomic affinity in these plants. Congeners may respond similarly in terms of reproductive output under global change, but fitness and prognoses of population persistence and evolutionary performance can be inferred only rarely from examination of vegetative characters alone.     

Effects of elevated CO2 on five plant-aphid interactions

We investigated interactions between five species of phloem-feeding aphids (Homoptera: Aphididae) and their host plants at elevated CO₂; Acyrthosiphon pisum (Harris) on Vicia faba L., Aphis nerii Boyer de Fonscolombe on Asclepias syriaca L., Aphis oenotherae Oestlund on Oenothera biennis L., Aulacorthum solani (Kaltenbach) on Nicotiana sylvestris Speg. & Comes and Myzus persicae (Sulzer) on Solanum dulcamara L. Host plants grown at elevated CO₂ generally had greater biomass, leaf area and C:N ratios than those grown at ambient CO₂, while plants with aphids had lower biomass and leaf area than those without aphids.The responses of aphid populations to elevated CO₂ were species-specific with one species increasing (M. persicae), one decreasing (A. pisum), and the other three being unaffected. CO₂ treatment did not affect the proportion of alate individuals produced. In general, aphid abundance was not significantly related to foliar nitrogen concentration.We performed separate analyses to test whether either aphid presence or aphid abundance modified the response of host plants to elevated CO₂. In terms of aphid presence, only three of the potential 15 interactions (five aphid species x three plant traits) were significant; A. solani slightly modified the response of the plant biomass to elevated CO₂ and M. persicae affected the response of leaf area and allocation. In terms of aphid abundance, only two of the potential 15 interactions were significant with A. nerii modifying the plant response to CO₂ in terms of total leaf area and allocation.We conclude that, in contrast to other insect groups such as leaf chewers, populations of most phloem-feeders may not be negatively affected by increased CO₂ concentrations in the future. The reasons for this difference include the possibility that aphids may be able to compensate for changes in host plant quality by altering feeding behaviour or by synthesizing amino acids. In addition, there is little evidence that aphid herbivory, even at high levels, will substantially modify the response of plants to elevated CO₂.     

Gas exchange of Ginkgo biloba leaves at different CO2 concentration levels

One and a half year-old Ginkgo saplings were grown for 2 years in 7 litre pots with medium fertile soil at ambient air CO₂ concentration and at 700 μmol mol⁻¹ CO₂ in temperature and humidity-controlled cabinets standing in the field. In the middle of the 2nd season of CO₂ enrichment, CO₂ exchange and transpiration in response to CO₂ concentration was measured with a mini-cuvette system. In addition, the same measurements were conducted in the crown of one 60-year-old tree in the field. Number of leaves/tree was enhanced by elevated CO₂ and specific leaf area decreased significantly.CO₂ compensation points were reached at 75–84 μmol mol⁻¹ CO₂. Gas exchange of Ginkgo saplings reacted more intensively upon CO₂ than those of the adult Ginkgo. On an average, stomatal conductance decreased by 30% as CO₂ concentration increased from 30 to 1000 μmol mol⁻¹ CO₂. Water use efficiency of net photosynthesis was positively correlated with CO₂ concentration levels. Saturation of net photosynthesis and lowest level of stomatal conductance was reached by the leaves of Ginkgo saplings at >1000 μmol mol⁻¹ CO₂. Acclimation of leaf net CO₂ assimilation to the elevated CO₂ concentration at growth occurred after 2 years of exposure. Maximum of net CO₂ assimilation was 56% higher at ambient air CO₂ concentration than at 700 μmol mol⁻¹ CO₂.