Oceanic protists with different forms of acquired phototrophy display contrasting biogeographies and abundance.

This first comprehensive analysis of the global biogeography of marine protistan plankton with acquired phototrophy shows these mixotrophic organisms to be ubiquitous and abundant; however, their biogeography differs markedly between different functional groups. These mixotrophs, lacking a constitutive capacity for photosynthesis (i.e. non-constitutive mixotrophs, NCMs), acquire their phototrophic potential through either integration of prey-plastids or through endosymbiotic associations with photosynthetic microbes. Analysis of field data reveals that 40-60% of plankton traditionally labelled as (non-phototrophic) microzooplankton are actually NCMs, employing acquired phototrophy in addition to phagotrophy. Specialist NCMs acquire chloroplasts or endosymbionts from specific prey, while generalist NCMs obtain chloroplasts from a variety of prey. These contrasting functional types of NCMs exhibit distinct seasonal and spatial global distribution patterns. Mixotrophs reliant on 'stolen' chloroplasts, controlled by prey diversity and abundance, dominate in high-biomass areas. Mixotrophs harbouring intact symbionts are present in all waters and dominate particularly in oligotrophic open ocean systems. The contrasting temporal and spatial patterns of distribution of different mixotroph functional types across the oceanic provinces, as revealed in this study, challenges traditional interpretations of marine food web structures. Mixotrophs with acquired phototrophy (NCMs) warrant greater recognition in marine research.

Stomatal vs. genome size in angiosperms: the somatic tail wagging the genomic dog?

BACKGROUND AND AIMS: Genome size is a function, and the product, of cell volume. As such it is contingent on ecological circumstance. The nature of 'this ecological circumstance' is, however, hotly debated. Here, we investigate for angiosperms whether stomatal size may be this 'missing link': the primary determinant of genome size. Stomata are crucial for photosynthesis and their size affects functional efficiency. METHODS: Stomatal and leaf characteristics were measured for 1442 species from Argentina, Iran, Spain and the UK and, using PCA, some emergent ecological and taxonomic patterns identified. Subsequently, an assessment of the relationship between genome-size values obtained from the Plant DNA C-values database and measurements of stomatal size was carried out. KEY RESULTS: Stomatal size is an ecologically important attribute. It varies with life-history (woody species < herbaceous species < vernal geophytes) and contributes to ecologically and physiologically important axes of leaf specialization. Moreover, it is positively correlated with genome size across a wide range of major taxa. CONCLUSIONS: Stomatal size predicts genome size within angiosperms. Correlation is not, however, proof of causality and here our interpretation is hampered by unexpected deficiencies in the scientific literature. Firstly, there are discrepancies between our own observations and established ideas about the ecological significance of stomatal size; very large stomata, theoretically facilitating photosynthesis in deep shade, were, in this study (and in other studies), primarily associated with vernal geophytes of unshaded habitats. Secondly, the lower size limit at which stomata can function efficiently, and the ecological circumstances under which these minute stomata might occur, have not been satisfactorally resolved. Thus, our hypothesis, that the optimization of stomatal size for functional efficiency is a major ecological determinant of genome size, remains unproven.

Influence on photosynthesis of starlight, moonlight, planetlight, and light pollution (reflections on photosynthetically active radiation in the universe).

Photosynthesis on Earth can occur in a diversity of organisms in the photosynthetically active radiation (PAR) range of 10 nmol of photons m(-2) s(-1) to 8 mmol of photons m(-2) s(-1). Similar considerations would probably apply to photosynthetic organisms on Earth-like planets (ELPs) in the continuously habitable zone of other stars. On Earth, starlight PAR is inadequate for photosynthetically supported growth. An increase in starlight even to reach the minimum theoretical levels to allow for photosynthesis would require a universe that was approximately ten million times older, or with a ten million times greater density of stars, than is the case for the present universe. Photosynthesis on an ELP using PAR reflected from a natural satellite with the same size as our Moon, but at the Roche limit, could support a low rate of photosynthesis at full Moon. Photosynthesis on an ELP-like satellite of a Jupiter-sized planet using light reflected from the planet could be almost 1% of the rate in full sunlight on Earth when the planet was full. These potential contributions to photosynthesis require that the contribution is compared with the rate of photosynthesis driven by direct radiation from the star. Light pollution on Earth only energizes photosynthesis by organisms that are very close to the light source. However, effects of light pollution on photosynthesis can be more widespread if the photosynthetic canopy is retained for more of the year, caused by effects on photoperiodism, with implications for the influence of civilizations on photosynthesis.

Significance of epidermal fusion and intercalary growth for angiosperm evolution.

The ancestral angiosperm flower probably had many separate elements in each floral whorl (sepals, petals, stamens and carpels). Derived character states include "fusion" of elements within a whorl (cohesion) and fusion between whorls (adhesion), as well as epigyny and the emergence of the other floral elements from the apex of the fused carpels. This article considers the roles of epidermal fusion and intercalary growth in the phylogeny and ontogeny of fused floral elements, and the importance of fusion for angiosperm evolution.

Ion transport in Hydrodictyon africanum.

The concentrations of K, Na, and Cl in the cytoplasm and vacuole, the tracer fluxes of these ions into and out of the cenocyte, and the electrical potential difference between bathing solution and vacuole and cytoplasm, have been measured in Hydrodictyon africanum. If the ions were acted on solely by passive electrochemical forces, a net efflux of K and Cl and a net influx of Na would be expected. Tracer fluxes indicate a net influx of K and Cl and efflux of Na in the light; these net fluxes are consequently active, with an obligate link to metabolism. The effects of darkness and low temperature indicate that most of the tracer K and Cl influx and Na efflux are linked to metabolism, while the corresponding tracer fluxes in the direction of the free energy gradient are not. Ouabain specifically inhibits the metabolically linked portions of tracer K influx and Na efflux. Alterations in the external K concentration have similar effects on metabolically mediated K influx and Na efflux. It would appear that K influx and Na efflux are linked, at least in the light.

Light stimulation of active transport in Hydrodictyon africanum.

The mechanism of light stimulation of active K and Cl influx and active Na efflux, in Hydrodictyon africanum has been investigated using different wavelengths of red light and different gas mixtures, and the inhibitors DCMU and CCCP. The active Cl influx requires photosystem 2, since its relative quantal efficiency falls with increasing wavelength of red light, and it is as sensitive to the inhibitor DCMU as is photosynthesis; it is relatively insensitive to the uncoupler CCCP. The active K influx and active Na efflux are inhibited by CCCP, but the relative quantal efficiency of these processes increases with increasing wavelength of red light, and they are relatively insensitive to DCMU. These cation fluxes can be supported by cyclic photophosphorylation, whereas Cl influx needs photosystem 2 but probably not ATP.

Iron stress restricts photosynthetic intersystem electron transport in Synechococcus sp. PCC 7942.

Although exposure of Synechococcus sp. PCC 7942 to iron stress induced the accumulation of the isiA gene product (CP43') compared with non-stressed controls, immunodetection of the N-terminus of cytochrome (Cyt) f indicated that iron stress not only reduced the content of the 40 kDa, heme-binding, Cyt f polypeptide by 32% but it also specifically induced the accumulation of a new, 23 kDa, non-heme-binding, putative Cyt f polypeptide. Concomitantly, iron stress restricted intersystem electron transport based on the in vivo reduction of P700(+), monitored as delta A(820)/A(820) in the presence and absence of electron transport inhibitors, as well as the inhibition of the Emerson enhancement effect on O(2) evolution. However, iron stress appeared to be associated with enhanced rates of PS I cyclic electron transport, low rates of PS I-driven photoreduction of NADP(+) but comparable rates for PS II+PS I photoreduction of NADP(+) relative to controls. We hypothesize that Synechococcus sp. PCC 7942 exhibits a dynamic capacity to uncouple PS II and PS I electron transport, which may allow for the higher than expected growth rates observed during iron stress.

Effect of elevated CO2 on the stomatal distribution and leaf physiology of Alnus glutinosa.

Variation in stomatal development and physiology of mature leaves from Alnus glutinosa plants grown under reference (current ambient, 360 µmol mol-1 CO2 ) and double ambient (720 µmol mol-1 CO2 ) carbon dioxide (CO2 ) mole fractions is assessed in terms of relative plant growth, stomatal characters (i.e. stomatal index and density) and leaf photosynthetic characters. This is the first study to consider the effects of elevated CO2 concentration on the distribution of stomata and epidermal cells across the whole leaf and to try to ascertain the cause of intraleaf variation. In general, a doubling of the atmospheric CO2 concentration enhanced plant growth and significantly increased stomatal index. However, there was no significant change in relative stomatal density. Under elevated CO2 concentration there was a significant decrease in stomatal conductance and an increase in assimilation rate. However, no significant differences were found for the maximum rate of carboxylation (Vcmax ) and the light saturated rate of electron transport (Jmax ) between the control and elevated CO2 treatment.

Variations of the natural abundances of nitrogen and carbon isotopes in Triticum aestivum, with special reference to phloem and xylem exudates.

This work explored whether the natural abundances of carbon and nitrogen isotopes could be used to describe the movement of C and X within wheat plants; we also considered whether isotopic analyses of aphids or their honeydew would substitute for direct analysis of phloem exudate. The δ13 C of ears and roots (sinks) most closely matched those of the sugars + organic acids fraction (sources) in both growth stages; phloem δ13 C matched that of leaf blade sugars. Xylem exudate δ13 C matched no other putative (and measured) source in the ear-forming stage and matched that of whole roots and ears in the grain-filling stage. The δ15 N of grain and roots (sinks) resembled that of leaf amino acids (sources) in the ear-forming stage. In the gram-filling stage, ear δ15 N continued to resemble that of leaf amino acids, and δ15 N of roots most closely resembled that of whole leaves. In the grain-filling stage, phloem δ15 N fell between that of leaf blade amino acids and that of whole leaves and was 15 X-depleted relative to internal and external NO, -N. In both growth stages, xylem exudate δ15 N was less than that of soil NO3 - -N and more than that of residual soil N after mineral N extraction. The isotopic values are generally in agreement with data from other approaches, such as isotope labelling; they show NO3 - -N reduction in both shoots and roots of wheat and significant N recycling (root-shoot-phloem-root) and C movement. Aphids might serve as a substitute for isotopic analysis of phloem δ15 N. having the same value as their food source. Their excreta was 15 N-enriched relative to phloem.

Inorganic carbon acquisition by aquatic photolithoatrophs of the Dighty Burn, Angus, U.K.: uses and limitations of natural abundance measurements of carbon isotopes.

The 13 C/12 C ratio (expressed as δ13 C) of benrhic photolithotrophs. in the Dighn Water (= Burn) were measured fur comparison with that of the potential inorganic carhun sources. CO2 and HCO3 - , in the Burn. The Burn water contains an average of 65.7 mmol m-3 CO2 with δ13 C of -14.7% and 1600 mmol m-3 HCO3 - with δ13 C of -4.%. δ13 C values of riparian vegetation were also measured as contributors, after respiration in the soil or the Burn, to the δ13 C of inorganic carbon in the Burn. The potential range of differences in 13 C/12C between dissolved CO2 and plant organic C is set by the intrinsic 13 c/12 C discrimination (α value) in CO2 fixation by Rubisco. Main results and conclusions are. as follows, (i) A literature survey suggests that there is no convincing evidence that the α, Values (rate constant for 12 CO2 fixation relative to that for 13 CO2 fixation by Rubisco in the absence of CO2 transport limitation) for the'lower plants'in the Burn (diatoms, green and red algae, mosses) are significantly different from the well-established αp values for the flowering plum enzyme. (ii) In confirmation of earlier work, the semi-erect 'streamer'gametophytes of the red alga Lemanea mamillosa and the moss Fontinalis antipyetica have δ13 C values which can only be interpreted in terms of diffusive CO2 entry with minimal limitation of photosynthesis by CO- diffusion, (iii) The serui-erect grren alga Cladophora glomerata and the flowering plant Ranunculus penicillatus ssp. pseudofluitons (formerly var. calcareus) are- both able to use HCO3 - . Their δ13 C values indicate that, if the HCO3 - -use system does not (as is likely) discriminate significantly between 13 C and 12 C, then a substantial fraction of the inorganic C made available to Rubisco must return to the medium, carrying 13 C-inorganic C not fixed by Rubisco. (iv) Two sets of δ13 C data from different hydrodynamic regimes distance from leading edge of a flat stone; different size of thalli) show that the attainable differences in situ in thickness of the diffusion boundary layer do not alter the fractional limitation of photosynthesis of Cladophora by external diffusion of inorganic C, considered with HCO3 use. (vi) The entrusting red alga Hildenbrandia rivularis has a δ13 C value suggestive of CO2 as the inorganic C source, but not entirely ruling nut HCO3 - . Marine species of both Hildenbrundia and Cladophora have δ13 C values which, even when corrected for source inorganic C δ13 C values, are 10%, more positive than the freshwater species. (vii) Mats of pennate diatoms were shown by pH-drift to by able to use HCO3 - ; the relatively high (i.e. not very negative) δ12 C value of these mats could relate to a relatively'non-leaky'HCO3 - aequisition mechanism and/or to limitation by external diffusion (e.g. through the mat).

Altritol synthesis by Notheia anomala.

The linear hexitol altritol has only been identified in six genera, all of them in the order Fucales of the brown algae. Five of these genera are closely related according to molecular phylogenetic and other data, while the sixth (Notheia) is an obligate epiphyte on two other altritol-containing genera with which it is symphanic. The possibility that Notheia obtains altritol from the algae on which it is epiphytic rather than by synthesizing altritol independently was investigated by supplying 13C-inorganic carbon in the light followed by mass spectrometric and nuclear magnetic resonance analysis. Notheia separated from the phorophyte Hormosira during exposure to 13C showed 13C enrichment in both altritol and mannitol, while the Hormosira only showed significant labelling of mannitol. These data show that altritol can be synthesized by Notheia, with implications for the number of gains and losses of the capacity to synthesize altritol in the evolution of the Fucales.

The acquisition of inorganic carbon by four red macroalgae.

Photosynthesis was studied in four species of red marine macroalgae: Palmaria palmata, Laurencia pinnatifida, Lomentaria articulata and Delesseria sanguinea. The rate of O2 evolution for submersed photosynthesis was measured as a function of incident photon flux density at normal pH and inorganic carbon concentration (pH 8.0, 2 mol m-3), and as a function of inorganic carbon concentration at pH 8.0 at saturating and at limiting photon flux density. The rate of CO2 uptake was measured for emersed photosynthesis as a function of CO2 partial pressure at saturating photon flux density. Previous pH-drift results suggest that Palmaria and Laurencia are able to use HCO inf3sup- as well as CO2 whereas Lomentaria and Delesseria are restricted to CO2. None of the algae are saturated by 2 mol m-3 inorganic carbon at high light (400 µmol m-2 s-1) but are saturated at low light (35 µmol m-2 s-1). The inorganic C concentration at which half the light-saturated rate of O2 evolution is achieved is higher for Palmaria and Laurencia (1.51 and 1.85 mol m-3) than for Lomentaria and Delesseria (0.772 and 0.841 mol m-3). The lower values for the latter two species could reflect their putative restriction to CO2. If expressed in terms of CO2, the half-saturation values yield 7.2 and 7.8 mmol m-3 respectively, which are very similar to values obtained previously during pH-drift experiments but at lower concentrations of HCO inf3sup- , consistent with restriction to CO2. The photosynthetic conductance (m s-1), calculated from the initial slope for photosynthesis at low concentrations of inorganic carbon, correlates with the suggested ability to extract inorganic carbon based on pH-drift results. Calculations made assuming that CO2 is the only species diffusing across the boundary layer are consistent with boundary layer thicknesses of 20 and 19 µm for Lomentaria and Delesseria respectively, which is feasible given the rapid water movement in the experiments. For Laurencia however, an unreasonably small boundary layer thickness of 6 µm is necessary to explain the flux, which indicates co-diffusion by HCO inf3sup- . In the apparent absence of external carbonic anhydrase, direct uptake of HCO inf3sup- , rather than external conversion to CO2 is indicated in this species. In air, the CO2 concentration at which photosynthesis is half-maximal increases in the same order as the ability to raise pH in drift experiments. At 21 kPa the CO2 compensation partial pressures for Palmaria and Laurencia at 0.56 and 1.3 Pa are low enough to suggest a carbon-concentrating mechanism is operating, while those of Lomentaria at 1.8 Pa and particularly that of Delesseria at 4.5 Pa could be explained without a carbon-concentrating mechanism. The algae tested (all except Delesseria) showed more O2 evolution than could be accounted for with a photosynthetic quotient of 1.0 and uncatalysed conversion of HCO inf3sup- to CO2 outside the cell in high light at pH 8.0 when high algal fresh weight per unit medium was used. These results are concordant with other data suggesting use of HCO inf3sup- by Palmaria and Laurencia, but discordant with the rest of the available information in indicating use of HCO inf3sup- by Lomentaria. The reason for this is unclear. The lightsaturated rate of O2 evolution on an algal area basis and the photon flux density needed to saturate photosynthesis were related partly to the habitat from which the seaweeds were collected, but more strongly to the ability to use HCO inf3sup- . Values for the two users of HCO inf3sup- , Palmaria (population used was intertidal; also occurs subtidally) and Laurencia (intertidal/shaded intertidal), were greater than for Lomentaria (shaded intertidal), which was greater than Delesseria (subtidal), both of which are believed to be restricted to CO2. In accordance with earlier δ13C data and, for Delesseria, estimates of the achieved growth rates in situ, carbon is likely to be saturating and use of HCO inf3sup- is unlikely to occur in the normal low-light habitats of Lomentaria and Delesseria. Analysis of N-use efficiencies show that they are closer to the low-CO2-affinity Laminariales than the high-CO2-affinity Fucaceae.

Discrimination between12C and13C by marine plants.

The natural abundance13C/12C ratios (as δ13C) of organic matter of marine macroalgae from Fife and Angus (East Scotland) were measured for comparison with the species' ability to use CO2 and HCO 3- for photosynthesis, as deduced from previously published pH-drift measurements. There was a clear difference in δ13C values for species able or unable to use HCO 3- . Six species of Chlorophyta, 12 species of Phaeophyta and 8 species of Rhodophyta that the pH-drift data suggested could use HCO 3- had δ13C values in the range -8.81‰ to -22.55‰. A further 6 species of Rhodophyta which the pH-drift data suggested could only use CO2 had δ13C values in the range -29.90‰ to-34.51‰. One of these six species (Lomentaria articulata) is intertidal; the other five are subtidal and so have no access to atmospheric CO2 to complicate the analysis. For these species, calculations based on the measured δ13C of the algae, the δ13C of CO2 in seawater, and the known13C/12C discrimination of CO2 diffusion and RUBISCO carboxylation suggest that only 15-21% of the limitation to photosynthesisin situ results from CO2 diffusion from the bulk medium to the plastids; the remaining 79-85% is associated with carboxylation reactions (and, via feedback effects, down-stream processes). This analysis has been extended for one of these five species,Delesseria sanguinea, by incorporating data onin situ specific growth rates, respiratory rates measured in the laboratory, and applying Fick's law of diffusion to calculate a boundary layer thickness of 17-24 µm. This value is reasonable for aDelesseria sanguinea frondin situ. For HCO 3- -using marine macroalgae the range of δ13C values measured can be accommodated by a CO2 efflux from algal cells which range from 0.306 of the gross HCO 3- influx forEnteromorpha intestinalis (δ13C=-8.81‰) in a rockpool to 0.787 forChondrus crispus (δ13C=-22.55‰). The relatively high computed CO2 efflux for those HCO 3- -users with the more negative δ13C values implies a relatively high photon cost of C assimilation; the observed photon costs can be accommodated by assuming coupled, energy-independent inorganic carbon influx and efflux. The observed δ13C values are also interpreted in terms of water movement regimes and obtaining CO2 from the atmosphere. Published δ13C values for freshwater macrophytes were compared with the ability of the species to use CO2 and HCO 3- and again there was an apparent separation in δ13C values for these two groups. δ13C values obtained for marine macroalgae for which no pH-drift data are available permit predictions, as yet untested, as to whether they use predominantly CO2 or HCO 3.

Inorganic carbon assimilation in the Isoetids, Isoetes lacustris L. and Lobelia dortmanna L.

The inorganic carbon fixation patterns of Isoetes lacustris and Lobelia dortmanna from an oligotrophic Scottish loch have been examined by following titratable acidity changes in plant sap and light/dark 14CO2 incorporation by roots and shoots. The diurnal pattern of titratable acidity changes in I. lacustris suggests crassulacean acid metabolism (CAM) while the lack of any change in titratable acidity in L. dortmanna suggests C3 metabolism. Of the carbon fixed by L. dortmanna, 99.9% was taken up through the roots and fixation occurred primarily during the day. In Isoetes, CO2 was taken up by both roots and shoots and during both day and night. Regardless of the site of CO2 uptake, fixation occurred only in the shoots of both plants. Analysis of carbon isotope ratios of plant organic material was used to further investigate the photosynthetic mechanisms of these Isoetids. Considering the absence of a nighttime peak in titratable acidity in L. dortmanna, the Δ13C (Δ=δ13C plant-δ13C source) value of the shoots of L. dortmanna (-14.2‰) is indicative of C3 photosynthesis limited by the rate of CO2 diffusion. The less negative Δ of I. lacustris (-6.0‰) is consistent with both dark acidification of CAM and CO2 limited C3 photosynthesis. This is in contrast to the terrestrial Isoetes durieui which is shown to have a Δ value which is similar to a terrestrial C3 plant. The carbon fixation patterns of these Isoetids suggest that the CO2 concentration in the loch may be growth limiting, and that root uptake and/or dark acidification are means of optimising CO2 supply. However, in view of the relatively high levels of CO2 in sediment and bulk water, it is suggested that low levels of nutrients may also limit growth in these plants.

Research note: cardiovascular effects of neurotensin in anesthetized White Leghorn hens.

Solutions of neurotensin (NT) at 0, 12, 60, 120, and 600 nM were infused i.v. into anesthetized, 16- to 34-wk-old hens at 60 microL/kg per minute for 30 min. These infusates increased plasma NT concentrations to steady state values of about .1, .2, 1, 2, and 10 times the postprandial concentrations of NT in hepatic portal blood, respectively. None of these concentrations significantly affected heart rate or arterial (carotid) or central venous (jugular) blood pressure.

Life and light: exotic photosynthesis in binary and multiple-star systems.

The potential for Earth-like planets within binary/multiple-star systems to host photosynthetic life was evaluated by modeling the levels of photosynthetically active radiation (PAR) such planets receive. Combinations of M and G stars in (i) close-binary systems; (ii) wide-binary systems, and (iii) three-star systems were investigated, and a range of stable radiation environments were found to be possible. These environmental conditions allow for the possibility of familiar, but also more exotic, forms of photosynthetic life, such as IR photosynthesizers and organisms that are specialized for specific spectral niches.

Biomineralization by photosynthetic organisms: evidence of coevolution of the organisms and their environment?

Biomineralization is widespread among photosynthetic organisms in the ocean, in inland waters and on land. The most quantitatively important biogeochemical role of land plants today in biomineralization is silica deposition in vascular plants, especially grasses. Terrestrial plants also increase the rate of weathering, providing the soluble substrates for biomineralization on land and in water bodies, a role that has had global biogeochemical impacts since the Devonian. The dominant photosynthetic biomineralizers in today's ocean are diatoms and radiolarians depositing silica and coccolithophores and foraminifera depositing calcium carbonate. Abiotic precipitation of silica from supersaturated seawater in the Precambrian preceded intracellular silicification dominated by sponges, then radiolarians and finally diatoms, with successive declines in the silicic acid concentration in the surface ocean, resulting in some decreases in the extent of silicification and, probably, increases in the silicic acid affinity of the active influx mechanisms. Calcium and bicarbonate concentrations in the surface ocean have generally been supersaturating with respect to the three common calcium carbonate biominerals through geological time, allowing external calcification as well as calcification in compartments within cells or organisms. The forms of calcium carbonate in biominerals, and presumably the evolution of the organisms that produce them, have been influenced by abiotic variations in calcium and magnesium concentrations in seawater, and calcium carbonate deposition has probably also been influenced by carbon dioxide concentration whose variations are in part biologically determined. Overall, there has been less biological feedback on the availability of substrates for calcification than is the case for silicification.