CO2 acquisition in Chlamydomonas acidophila is influenced mainly by CO2, not phosphorus,availability

The extremophilic green microalga Chlamydomonas acidophila grows in very acidic waters (pH 2.3–3.4), where CO₂ is the sole inorganic carbon source. Previous work has revealed that the species can accumulate inorganic carbon (Ci) and exhibits high affinity CO₂ utilization under low-CO₂ (air-equilibrium) conditions, similar to organisms with an active CO₂ concentrating mechanism (CCM), whereas both processes are down-regulated under high CO₂ (4.5 % CO₂) conditions. Responses of this species to phosphorus (Pi)-limited conditions suggested a contrasting regulation of the CCM characteristics. Therefore, we measured external carbonic anhydrase (CA_ext) activities and protein expression (CAH1), the internal pH, Ci accumulation, and CO₂-utilization in cells adapted to high or low CO₂ under Pi-replete and Pi-limited conditions. Results reveal that C. acidophila expressed CA_ext activity and expressed a protein cross-reacting with CAH1 (the CA_ext from Chlamydomonas reinhardtii). Although the function of this CA remains unclear, CA_ext activity and high affinity CO₂ utilization were the highest under low CO₂ conditions. C. acidophila accumulated Ci and expressed the CAH1 protein under all conditions tested, and C. reinhardtii also contained substantial amounts of CAH1 protein under Pi-limitation. In conclusion, Ci utilization is optimized in C. acidophila under ecologically relevant conditions, which may enable optimal survival in its extreme Ci- and Pi-limited habitat. The exact physiological and biochemical acclimation remains to be further studied.     

Bicarbonate uptake via an anion exchange protein is the main mechanism of inorganic carbon acquisition by the giant kelp Macrocystis pyrifera (Laminariales,Phaeophyceae) under variable pH

Macrocystis pyrifera is a widely distributed, highly productive, seaweed. It is known to use bicarbonate (HCO₃^?) from seawater in photosynthesis and the main mechanism of utilization is attributed to the external catalyzed dehydration of HCO₃^? by the surface‐bound enzyme carbonic anhydrase (CA_ext). Here, we examined other putative HCO₃^? uptake mechanisms in M. pyrifera under pH_T 9.00 (HCO₃^?: CO₂ = 940:1) and pH_T 7.65 (HCO₃^?: CO₂ = 51:1). Rates of photosynthesis, and internal CA (CA_int) and CA_ext activity were measured following the application of AZ which inhibits CA_ext, and DIDS which inhibits a different HCO₃^? uptake system, via an anion exchange (AE) protein. We found that the main mechanism of HCO₃^? uptake by M. pyrifera is via an AE protein, regardless of the HCO₃^?: CO₂ ratio, with CA_ext making little contribution. Inhibiting the AE protein led to a 55%–65% decrease in photosynthetic rates. Inhibiting both the AE protein and CA_ext at pH_T 9.00 led to 80%–100% inhibition of photosynthesis, whereas at pH_T 7.65, passive CO₂ diffusion supported 33% of photosynthesis. CA_int was active at pH_T 7.65 and 9.00, and activity was always higher than CA_ext, because of its role in dehydrating HCO₃^? to supply CO₂ to RuBisCO. Interestingly, the main mechanism of HCO₃^? uptake in M. pyrifera was different than that in other Laminariales studied (CA_ext‐catalyzed reaction) and we suggest that species‐specific knowledge of carbon uptake mechanisms is required in order to elucidate how seaweeds might respond to future changes in HCO₃^?:CO₂ due to ocean acidification.     

The role of external carbonic anhydrase in photosynthesis during growth of the marine diatom Chaetoceros muelleri

Carbon dioxide concentrating mechanisms (CCMs) act to improve the supply of CO₂ at the active site of ribulose‐1,5‐bisphosphate carboxylase/oxygenase. There is substantial evidence that in some microalgal species CCMs involve an external carbonic anhydrase (CA_ext) and that CA_ext activity is induced by low CO₂ concentrations in the growth medium. However, much of this work has been conducted on cells adapted to air‐equilibrium concentrations of CO₂, rather than to changing CO₂ conditions caused by growing microalgal populations. We investigated the role of CA_ext in inorganic carbon (C_i) acquisition and photosynthesis at three sampling points during the growth cycle of the cosmopolitan marine diatom Chaetoceros muelleri. We observed that CA_ext activity increased with decreasing C_i, particularly CO₂, concentration, supporting the idea that CA_ext is modulated by external CO₂ concentration. Additionally, we found that the contribution of CA_ext activity to carbon acquisition for photosynthesis varies over time, increasing between the first and second sampling points before decreasing at the last sampling point, where external pH was high. Lastly, decreases in maximum quantum yield of photosystem II (F_v/F_m), chlorophyll, maximum relative electron transport rate, light harvesting efficiency (α) and maximum rates of C_i‐ saturated photosynthesis (V_max) were observed over time. Despite this decrease in photosynthetic capacity an up‐regulation of CCM activity, indicated by a decreasing half‐saturation constant for CO₂ (K_0.5CO₂), occurred over time. The flexibility of the CCM during the course of growth in C. muelleri may contribute to the reported dominance and persistence of this species in phytoplankton blooms.     

The induction of inorganic carbon transport and external carbonic anhydrase in Chlamydomonas reinhardtii is regulated by external CO2 concentration

Induction of the carbon concentrating mechanism (CCM) has been investigated during the acclimation of 5% CO₂‐grown Chlamydomonas reinhardtii 2137 mt + cells to well‐defined dissolved inorganic carbon (C_i) limited conditions. The CCM components investigated were active HCO₃^? transport, active CO₂ transport and extracellular carbonic anhydrase (CA_ext) activity. The CA_ext activity increased 10‐fold within 6 h of acclimation to 0·035% CO₂ and there was a further slight increase over the next 18 h. The CA_ext activity also increased substantially after an 8 h lag period during acclimation to air in darkness. Active CO₂ and HCO₃^? uptake by C. reinhardtii cells were induced within 2 h of acclimation to air, but active CO₂ transport was induced prior to active HCO₃^? transport. Similar results were obtained during acclimation to air in darkness. The critical C_i concentrations effecting the induction of active C_i transport and CA_ext activity were determined by allowing cells to acclimate to various inflow CO₂ concentrations in the range 0·035–0·84% at constant pH. The total C_i concentration eliciting the induction and repression of active C_i transport was higher during acclimation at pH 7·5 than at pH 5·5, but the external CO₂ concentration was the same at both pHs of acclimation. The concentration of external CO₂ required for the full induction and repression of C_i transport and CA_ext activity were 10 and 100 μM , respectively. The induction of CA_ext and active C_i transport are not correlated temporally, but are regulated by the same critical CO₂ concentration in the medium.     

Isotope Fractionation in Photosynthetic Bacteria during Carbon Dioxide Assimilation

The δ _PDB¹³C values have been determined for the cellular constituents and metabolic intermediates of autotrophically grown Chromatium vinosum. The isotopic composition of the HCO₃^- in the medium and the carbon isotopic composition of the bacterial cells change with the growth of the culture. The δ _PDB¹³C value of the HCO₃^- in the media changes from an initial value of −6.6‰ to +8.1‰ after 10 days of bacterial growth and the δ _PDB¹³C value of the bacterial cells change from −37.5‰ to −29.2‰ in the same period. The amount of carbon isotope fractionation during the synthesis of hexoses by the photoassimilation of CO₂ has a range of −15.5‰ at time zero to −22.0‰ after 10 days. This range of fractionation compares to the range of carbon isotope fractionation for the synthesis of sugars from CO₂ by ribulose 1,5-diphosphate carboxylase and the Calvin cycle.     

Responses to shading of naturalized and non-naturalized exotic woody species

Background and Aims

Recent studies have suggested that responses to shading gradients may play an important role in establishment success of exotic plants, but hitherto few studies have tested this. Therefore, a common-garden experiment was conducted using multiple Asian woody plant species that were introduced to Europe >100 years ago in order to test whether naturalized and non-naturalized species differ in their responses to shading. Specifically, a test was carried out to determine whether naturalized exotic woody species maintained better growth under shaded conditions, and whether they expressed greater (morphological and physiological) adaptive plasticity in response to shading, relative to non-naturalized species.

Methods

Nineteen naturalized and 19 non-naturalized exotic woody species were grown under five light levels ranging from 100 to 7 % of ambient light. For all plants, growth performance (i.e. biomass), morphological and CO₂ assimilation characteristics were measured. For the CO₂ assimilation characteristics, CO₂ assimilation rate was measured at 1200 μmol m^–2 s^–1 (i.e. saturated light intensity, A₁₂₀₀), 50 μmol m^–2 s^–1 (i.e. low light intensity, A₅₀) and 0 μmol m^–2 s^–1 (A₀, i.e. dark respiration).

Key Results

Overall, the naturalized and non-naturalized species did not differ greatly in biomass production and measured morphological and CO₂ assimilation characteristics across the light gradient. However, it was found that naturalized species grew taller and reduced total leaf area more than non-naturalized species in response to shading. It was also found that naturalized species were more capable of maintaining a high CO₂ assimilation rate at low light intensity (A₅₀) when grown under shading.

Conclusions

The results indicate that there is no clear evidence that the naturalized species possess a superior response to shading over non-naturalized species, at least not at the early stage of their growth. However, the higher CO₂ assimilation capacity of the naturalized species under low-light conditions might facilitate early growth and survival, and thereby ultimately favour their initial population establishment over the non-naturalized species.     

MORPHOLOGICAL AND PHYSIOLOGICAL EFFECTS IN PROBOSCIA ALATA (BACILLARIOPHYCEAE) GROWN UNDER DIFFERENT LIGHT AND CO2 CONDITIONS OF THE MODERN SOUTHERN OCEAN1

The combined effects of different light and aqueous CO₂ conditions were assessed for the Southern Ocean diatom Proboscia alata (Brightwell) Sundström in laboratory experiments. Selected culture conditions (light and CO_2(aq)) were representative for the natural ranges in the modern Southern Ocean. Light conditions were 40 (low) and 240 (high) μmol photons · m^?2 · s^?1. The three CO_2(aq) conditions ranged from 8 to 34 μmol · kg^?1 CO_2(aq) (equivalent to a pCO₂ from 137 to 598 μatm, respectively). Clear morphological changes were induced by these different CO_2(aq) conditions. Cells in low [CO_2(aq)] formed spirals, while many cells in high [CO_2(aq)] disintegrated. Cell size and volume were significantly affected by the different CO_2(aq) concentrations. Increasing CO_2(aq) concentrations led to an increase in particulate organic carbon concentrations per cell in the high light cultures, with exactly the opposite happening in the low light cultures. However, other parameters measured were not influenced by the range of CO_2(aq) treatments. This included growth rates, chlorophyll a concentration and photosynthetic yield (F_V/F_M). Different light treatments had a large effect on nutrient uptake. High light conditions caused an increased nutrient uptake rate compared to cells grown in low light conditions. Light and CO₂ conditions co‐determined in various ways the response of P. alata to changing environmental conditions. Overall P. alata appeared to be well adapted to the natural variability in light availability and CO_2(aq) concentration of the modern Southern Ocean. Nevertheless, our results showed that P. alata is susceptible to future changes in inorganic carbon concentrations in the Southern Ocean.     

Carbon Isotope Fractionation of 11 Acetogenic Strains Grown on H2 and CO2

Acetogenic bacteria are able to grow autotrophically on hydrogen and carbon dioxide by using the acetyl coenzyme A (acetyl-CoA) pathway. Acetate is the end product of this reaction. In contrast to the fermentative route of acetate production, which shows almost no fractionation of carbon isotopes, the acetyl-CoA pathway has been reported to exhibit a preference for light carbon. In Acetobacterium woodii the isotope fractionation factor (ε) for ¹³C and ¹²C has previously been reported to be ε = −58.6‰. To investigate whether such a strong fractionation is a general feature of acetogenic bacteria, we measured the stable carbon isotope fractionation factor of 10 acetogenic strains grown on H₂ and CO₂. The average fractionation factor was ε_TIC = −57.2‰ for utilization of total inorganic carbon and ε_acetate = −54.6‰ for the production of acetate. The strongest fractionation was found for Sporomusa sphaeroides (ε_TIC = −68.3‰), the lowest fractionation for Morella thermoacetica (ε_TIC = −38.2‰). To investigate the reproducibility of our measurements, we determined the fractionation factor of 21 biological replicates of Thermoanaerobacter kivui. In general, our study confirmed the strong fractionation of stable carbon during chemolithotrophic acetate formation in acetogenic bacteria. However, the specific characteristics of the bacterial strain, as well as the cultural conditions, may have a moderate influence on the overall fractionation.     

The role of extracellular carbonic anhydrase for accumulation of inorganic carbon in the green alga Chlamydomonas reinhardtii. A comparison between wild-type and cell-wall-less mutant cells

The role of extracellular carbonic anhydrase (CA_ex) for dissolved inorganic carbon (DIC) accumulation in the green alga Chlamydomonas reinhardtii was investigated. It was found that when algal cells were bubbled with ambient air, cell-wall-less mutant cells exhibited the same high photosynthetic affinity for CO₂ as wild-type cells despite a 10 times lower activity of CA^ex. It was also found that the affinity for CO₂ was further increased when the total DIC concentration of the algal medium was reduced from that in equilibrium with ambient air to even lower levels. This increased affinity was not correlated with any further increase in the CA_ex activity. Dextran-bound sulfonamide (DBS. 100 μM bound ligand) completely inhibited the activity of CA_ex in intact, low-DIC grown, wild-type cells, while photosynthesis at <2 μM CO₂(aq) proceeded at a far greater rate than could be maintained by CO₂ supplied from the spontaneous dehydration of HCO^?₃. DBS-inhibition of CA_ex, during the induction of the DIC-accumulating mechanism in previously high-DIC grown cells, only caused a 50% inhibition of photosynthesis at 10 μM CO₂(aq) after 1 h of low-DIC acclimation. It was also shown that 50 μM acetazolamide (AZ) inhibited photosynthesis at low DIC concentrations to a relatively higher degree than DBS, suggesting that AZ inhibited intracellular CA as well. Taken together, these results suggest that low-DIC grown cells of C. reinhardtii have the ability to transport HCO^?₃ across the plasma membrane in addition to the CA_ex-mediated, facilitated diffusion and/or transport of CO₂. It is also suggested that the relative importance of these two fluxes (CO₂ or HCO^?₃) is dependent on the growth and experimental conditions. Facilitated CO₂ uptake seems to be most prevalent, supported by HCO^?₃-transport under more or less extreme situations, such as a reduction of CO₂ to extremely low concentrations, leakage of CA_ex to the medium as in cultures of cell-wall-less mutant cells or when the activity of CA_ex has been artificially inhibited.     

Stable Carbon Isotope Fractionation by Sulfate-Reducing Bacteria

Biogeochemical transformations occurring in the anoxic zones of stratified sedimentary microbial communities can profoundly influence the isotopic and organic signatures preserved in the fossil record. Accordingly, we have determined carbon isotope discrimination that is associated with both heterotrophic and lithotrophic growth of pure cultures of sulfate-reducing bacteria (SRB). For heterotrophic-growth experiments, substrate consumption was monitored to completion. Sealed vessels containing SRB cultures were harvested at different time intervals, and δ¹³C values were determined for gaseous CO₂, organic substrates, and products such as biomass. For three of the four SRB, carbon isotope effects between the substrates, acetate or lactate and CO₂, and the cell biomass were small, ranging from 0 to 2‰. However, for Desulfotomaculum acetoxidans, the carbon incorporated into biomass was isotopically heavier than the available substrates by 8 to 9‰. SRB grown lithoautotrophically consumed less than 3% of the available CO₂ and exhibited substantial discrimination (calculated as isotope fractionation factors [α]), as follows: for Desulfobacterium autotrophicum, α values ranged from 1.0100 to 1.0123; for Desulfobacter hydrogenophilus, the α value was 0.0138, and for Desulfotomaculum acetoxidans, the α value was 1.0310. Mixotrophic growth of Desulfovibrio desulfuricans on acetate and CO₂ resulted in biomass with a δ¹³C composition intermediate to that of the substrates. The extent of fractionation depended on which enzymatic pathways were used, the direction in which the pathways operated, and the growth rate, but fractionation was not dependent on the growth phase. To the extent that environmental conditions affect the availability of organic substrates (e.g., acetate) and reducing power (e.g., H₂), ecological forces can also influence carbon isotope discrimination by SRB.     

Photosynthesis of Ulva sp. II. utilization of CO2 and HCO−3 when submerged

The photosynthetic capacity of submerged Ulva sp. when utilizing CO₂ and HCO^?₃ as exogenous carbon forms has been investigated and compared with ambient carbon concentrations in sea water. Saturating concentrations of HCO^{? ₃} and CO₂ were 1200 and 100 μM, respectively at saturating light, and photosynthetic rates under such conditions averaged 700 μmolO₂·gDW^?1 ·h^?1. The HCO^?₃ concentration of sea water (≈2500μM), was thus found to be saturating for photosynthesis of Ulva. At the CO₂ concentration of sea water (≈ 10 μM), the contribution of this carbon form to photosynthesis could be 27% at the most. Under conditions of slow water movement, the relative importance of CO₂ utilization would probably be minimized in favour of HCO^?₃ utilization. It is concluded that HCO^?₃ uptake is not limiting photosynthesis for Ulva under natural conditions.     

Enzymatic fractionation of carbon isotopes by phosphoenolpyruvate carboxylase from c(4) plants

The carbon atoms of glucose and malate in C₄ plants are 2 to 3‰ enriched in ¹²C with respect to atmospheric CO₂; whereas these intermediates in C₃ plants are 15 to 18‰ enriched with ¹²C with respect to atmospheric CO₂. The enzymatic synthesis of malate from phosphoenolpyruvate and bicarbonate in preparations of leaves of Sorghum bicolor, Haygrazer result in a carbon isotope fractionation of about 3‰. The enzymatic synthesis of phosphoglyceric acid from ribulose 1,5-diP and CO₂ in these preparations (contaminated with carbonic anhydrase) at 24 C and 37 C result in a carbon isotope fractionation of 33.7‰ and 18.3‰, respectively. These data are consistent with the conclusion that the small enrichment of ¹²C in the carbon atoms of malate and glucose (with respect to atmospheric CO₂) in leaves of Sorghum bicolor, Haygrazer occurs at the phosphoenolpyruvate carboxylase step.     

Carbon limitation enhances CO2 concentrating mechanism but reduces trichome size in Arthrospira platensis (cyanobacterium)

Arthrospira species grow well under highly enriched inorganic carbon concentrations, but little is known on the effects of inorganic carbon (Ci) limitation on its physiological performance. When Arthrospira platensis D-0083 was grown in a modified medium without NaHCO₃ under ambient air of 380 ppm CO₂, its trichomes became disassembled while the growth and photosynthetic rates were severely reduced. Phycocyanin and allophycocyanin contents decreased but the carotenoid content increased under the Ci limitation. Compared with the cells grown in Zarrouk medium, the trichomes grown under the Ci limitation increased their photosynthetic apparent affinity for Ci by about 14 times but photochemical quenching capacity was reduced. It appeared that A. platensis increased its CO₂ concentrating mechanism by inducing HCO₃ ^? transporters and reducing the trichome size which increased filamentous surface to volume ratio.     

Photosynthetic carbon metabolism of a marine grass

The δ¹³C value of a tropical marine grass Thalassia testudinum is −9.04‰. This value is similar to the δ¹³C value of terrestrial tropical grasses. The δ¹³C values of the organic acid fraction, the amino acid fraction, the sugar fraction, malic acid, and glucose are: −11.2‰, −13.1‰, −10.1‰, −11.1‰, and −11.5‰, respectively. The δ¹³C values of malic acid and glucose of Thalassia are similar to the δ¹³C values of these intermediates in sorghum leaves and attest to the presence of the photosynthetic C₄-dicarboxylic acid pathway in this marine grass. The inorganic HCO₃⁻ for the growth of the grass fluctuates between −6.7 to −2.7‰ during the day. If CO₂ fixation in Thalassia is catalyzed by phosphoenolpyruvate carboxylase (which would result in a −3‰ fractionation between HCO₃⁻ and malic acid), the predicted δ¹³C value for Thalassia would be −9.7 to −5.7‰. This range is close to the observed range of −12.6 to −7.8‰ for Thalassia and agree with the operation of the C₄-dicarboxylic acid pathway in this plant. The early products of the fixation of HCO₃⁻ in the leaf sections are malic acid and aspartic acid which are similar to the early products of CO₂ fixation in C₄ terrestrial plants.     

Carbon isotope fractionation by ribulose-1,5-bisophosphate carboxylase from various organisms

Carbon isotope fractionation by structurally and catalytically distinct ribulose-1,5-bisphosphate carboxylases from one eucaryotic and four procaryotic organisms has been measured under nitrogen. The average fractionation for 40 experiments was −34.1 ‰ with respect to the δ¹³C of the dissolved CO₂ used, although average fractionations for each enzyme varied slightly: spinach carboxylase, −36.5 ‰; Hydrogenomonas eutropha, −38.7 ‰; Agmenellum quadruplicatum, −32.2 ‰; Rhodospirillum rubrum, −32.1 ‰; Rhodopseudomonas sphaeroides peak I carboxylase, −31.4 ‰; and R. sphaeroides peak II carboxylase, −28.3 ‰. The carbon isotope fractionation value was largely independent of method of enzyme preparation, purity, or reaction temperature, but in the case of spinach ribulose-1,5-bisphosphate carboxylase fractionation, changing the metal cofactor used for enzyme activation had a distinct effect on the fractionation value. The fractionation value of −36.5 ‰ with Mg²⁺ as activator shifted to −29.9 ‰ with Ni²⁺ as activator and to −41.7 ‰ with Mn²⁺ as activator. These dramatic metal effects on carbon isotope fractionation may be useful in examining the catalytic site of the enzyme.     

Stable Water Use Efficiency of Tibetan Alpine Meadows in Past Half Century: Evidence from Wool δ13C Values

Understanding the influences of climatic changes on water use efficiency (WUE) of Tibetan alpine meadows is important for predicting their long-term net primary productivity (NPP) because they are considered very sensitive to climate change. Here, we collected wool materials produced from 1962 to 2010 and investigated the long-term WUE of an alpine meadow in Tibet on basis of the carbon isotope values of vegetation (δ ¹³C_veg). The values of δ ¹³C_veg decreased by 1.34‰ during 1962–2010, similar to changes in δ ¹³C values of atmospheric CO₂. Carbon isotope discrimination was highly variable and no trend was apparent in the past half century. Intrinsic water use efficiency (W _i) increased by 18 μmol·mol^–1 (approximately 23.5%) during 1962–2010 because the increase in the intercellular CO₂ concentration (46 μmol·mol^–1) was less than that in the atmospheric CO₂ concentration (C _a, 73 μmol·mol^–1). In addition, W _i increased significantly with increasing growing season temperature and C _a. However, effective water use efficiency (W _e) remained relatively stable, because of increasing vapor pressure deficit. C _a, precipitation, and growing season temperature collectively explained 45% of the variation of W _e. Our findings indicate that the W _e of alpine meadows in the Tibetan Plateau remained relatively stable by physiological adjustment to elevated C _a and growing season temperature. These findings improve our understanding and the capacity to predict NPP of these ecosystems under global change scenarios.     

Effects of CO2 and nitrogen supply on the biochemical composition of Ulva rigida with especial emphasis on lipid class analysis

Lipid class composition was analysed in the green macroalga Ulva rigida grown under normal (350 ppm) and high (10,000 ppm) CO₂ levels, and in nitrate saturated and nitrogen limited conditions. A new protocol for the extraction of lipids has been defined. Culture conditions altered the fate of assimilated carbon, and significant changes were observed in protein and total lipid content in particular. A CO₂-enriched atmosphere conditioned the effects of nitrogen limitation on lipid class composition, revealing deep qualitative changes in carbon metabolism. Triglycerides accumulated at high CO₂ and under nitrogen limitation, while chloroplast-related lipids showed an inverse response. Changes in phospholipids could be related to carbon availability as they did not respond to nitrogen limitation. The ratio sterols/acetone-mobile polar lipids followed a negative linear relation with the optimum quantum yield for photosynthetic electron transport (F_v/F_m), and was considered as an index of the «light status» of the cell. The specificity of the response of lipid classes to growth conditions in U. rigida emphasizes the potential role of lipid class analyses as a diagnostic tool for environmental stress.     

Signature Lipids and Stable Carbon Isotope Analyses of Octopus Spring Hyperthermophilic Communities Compared with Those of Aquificales Representatives

The molecular and isotopic compositions of lipid biomarkers of cultured Aquificales genera have been used to study the community and trophic structure of the hyperthermophilic pink streamers and vent biofilm from Octopus Spring. Thermocrinis ruber, Thermocrinis sp. strain HI 11/12, Hydrogenobacter thermophilus TK-6, Aquifex pyrophilus, and Aquifex aeolicus all contained glycerol-ether phospholipids as well as acyl glycerides. The n-C_20:1 and cy-C₂₁ fatty acids dominated all of the Aquificales, while the alkyl glycerol ethers were mainly C_18:0. These Aquificales biomarkers were major constituents of the lipid extracts of two Octopus Spring samples, a biofilm associated with the siliceous vent walls, and the well-known pink streamer community (PSC). Both the biofilm and the PSC contained mono- and dialkyl glycerol ethers in which C₁₈ and C₂₀ alkyl groups were prevalent. Phospholipid fatty acids included both the Aquificales n-C_20:1 and cy-C₂₁, plus a series of iso-branched fatty acids (i-C_15:0 to i-C_21:0), indicating an additional bacterial component. Biomass and lipids from the PSC were depleted in ¹³C relative to source water CO₂ by 10.9 and 17.2‰, respectively. The C_20–21 fatty acids of the PSC were less depleted than the iso-branched fatty acids, 18.4 and 22.6‰, respectively. The biomass of T. ruber grown on CO₂ was depleted in ¹³C by only 3.3‰ relative to C source. In contrast, biomass was depleted by 19.7‰ when formate was the C source. Independent of carbon source, T. ruber lipids were heavier than biomass (+1.3‰). The depletion in the C_20–21 fatty acids from the PSC indicates that Thermocrinis biomass must be similarly depleted and too light to be explained by growth on CO₂. Accordingly, Thermocrinis in the PSC is likely to have utilized formate, presumably generated in the spring source region.     

Adjustment to the light environment in small-statured forbs as a strategy for complementary resource use in mixtures of grassland species

Background and Aims

The biological mechanisms of niche complementarity allowing for a stable coexistence of a large number of species in a plant community are still poorly understood. This study investigated how small-statured forbs use environmental niches in light and CO₂ to explain their persistence in diverse temperate grasslands.

Methods

Light and CO₂ profiles and the corresponding leaf characteristics of seven small-statured forbs were measured in monocultures and a multi-species mixture within a biodiversity experiment (Jena Experiment) to assess their adjustment to growth conditions in the canopy.

Key Results

Environmental conditions near the ground varied throughout the season with a substantial CO₂ enrichment (>70 µmol mol⁻¹ at 2 cm, >20 µmol mol⁻¹ at 10 cm above soil surface) and a decrease in light transmittance (to <5 % deep in the canopy) with large standing biomass (>500 g d. wt m⁻²) in the multi-species assemblage. Leaf morphology, biochemistry and physiology of small-statured forbs adjusted to low light in the mixture compared with the monocultures. However, the net carbon assimilation balance during the period of low light only compensated the costs of maintenance respiration, while CO₂ enrichment near the ground did not allow for additional carbon gain. Close correlations of leaf mass per area with changes in light availability suggested that small-statured forbs are capable of adjusting to exploit seasonal niches with better light supply for growth and to maintain the carbon metabolism for survival if light transmittance is substantially reduced in multi-species assemblages.

Conclusions

This study shows that adjustment to a highly dynamic light environment is most important for spatial and seasonal niche separation of small-statured forb species in regularly mown, species-rich grasslands. The utilization of short-period CO₂ enrichment developing in dense vegetation close to the ground hardly improves their carbon balance and contributes little to species segregation along environmental niche axes.     

Priming of Soil Carbon Decomposition in Two Inner Mongolia Grassland Soils following Sheep Dung Addition: A Study Using 13C Natural Abundance Approach

To investigate the effect of sheep dung on soil carbon (C) sequestration, a 152 days incubation experiment was conducted with soils from two different Inner Mongolian grasslands, i.e. a Leymus chinensis dominated grassland representing the climax community (2.1% organic matter content) and a heavily degraded Artemisia frigida dominated community (1.3% organic matter content). Dung was collected from sheep either fed on L. chinensis (C₃ plant with δ¹³C = −26.8‰; dung δ¹³C = −26.2‰) or Cleistogenes squarrosa (C₄ plant with δ¹³C = −14.6‰; dung δ¹³C = −15.7‰). Fresh C₃ and C₄ sheep dung was mixed with the two grassland soils and incubated under controlled conditions for analysis of ¹³C-CO₂ emissions. Soil samples were taken at days 17, 43, 86, 127 and 152 after sheep dung addition to detect the δ¹³C signal in soil and dung components. Analysis revealed that 16.9% and 16.6% of the sheep dung C had decomposed, of which 3.5% and 2.8% was sequestrated in the soils of L. chinensis and A. frigida grasslands, respectively, while the remaining decomposed sheep dung was emitted as CO₂. The cumulative amounts of C respired from dung treated soils during 152 days were 7–8 times higher than in the un-amended controls. In both grassland soils, ca. 60% of the evolved CO₂ originated from the decomposing sheep dung and 40% from the native soil C. Priming effects of soil C decomposition were observed in both soils, i.e. 1.4 g and 1.6 g additional soil C kg⁻¹ dry soil had been emitted as CO₂ for the L. chinensis and A. frigida soils, respectively. Hence, the net C losses from L. chinensis and A. frigida soils were 0.6 g and 0.9 g C kg⁻¹ soil, which was 2.6% and 7.0% of the total C in L. chinensis and A. frigida grasslands soils, respectively. Our results suggest that grazing of degraded Inner Mongolian pastures may cause a net soil C loss due to the positive priming effect, thereby accelerating soil deterioration.