A CO2-enriched atmosphere improves in vitro growth of Brazilian ginseng [Pfaffia glomerata (Spreng.) Pedersen]

The aim of the present study was to evaluate the effects of forced ventilation and CO₂ enrichment (360 or 720 μmol mol^?1 CO₂) on the in vitro growth and development of Pfaffia glomerata, an endangered medicinal species, under photomixotrophic or photoautotrophic conditions. P. glomerata nodal segments showed substantial differences in growth, relative water content and water loss from leaves, photosynthetic pigments, stomatal density, and leaf anatomical characteristics under these different treatments. CO₂ enrichment led to increased photosynthetic pigments and reduced stomatal density of in vitro cultivated P. glomerata. A lack of sucrose in the culture medium increased 20-hydroxyecdysone levels, but the increase in CO₂ levels did not further elevate the accumulation of 20-hydroxyecdysone. All growth increased in a CO₂-enriched atmosphere. In addition, CO₂ enrichment, with or without sucrose, gave a lower relative water loss from leaves. This finding indicates that either a photoautotrophic or photomixotrophic system in a CO₂-enriched atmosphere may be suitable for large-scale propagation of this species.     

Photosynthetic refixing of CO2 is responsible for the apparent disparity between mitochondrial respiration in the light and in the dark

The net photosynthetic rate (P _N), the sample room CO₂ concentration (CO₂S) and the intercellular CO₂ concentration (C _i) in response to PAR, of C₃ (wheat and bean) and C₄ (maize and three-colored amaranth) plants were measured. Results showed that photorespiration (R _p) of wheat and bean could not occur at 2 % O₂. At 2 % O₂ and 0 μmol mol^?1 CO₂, P _N can be used to estimate the rate of mitochondrial respiration in the light (R _d). The R _d decreased with increasing PAR, and ranged between 3.20 and 2.09 μmol CO₂ m^?2 s^?1 in wheat. The trend was similar for bean (between 2.95 and 1.70 μmol CO₂ m^?2 s^?1), maize (between 2.27 and 0.62 μmol CO₂ m^?2 s^?1) and three-colored amaranth (between 1.37 and 0.49 μmol CO₂ m^?2 s^?1). The widely observed phenomenon of R _d being lower than R _n can be attributed to refixation, rather than light inhibition. For all plants tested, CO₂ recovery rates increased with increasing light intensity from 32 to 55 % (wheat), 29 to 59 % (bean), 54 to 87 % (maize) and 72 to 90 % (three-colored amaranth) at 50 and 2,000 μmol m^?2 s^?1, respectively.     

<Emphasis Type="Italic">Ex vitro</Emphasis> rooting using a mini growth chamber increases root induction and accelerates acclimatization of Kopyor coconut (<Emphasis Type="Italic">Cocos nucifera</Emphasis> L.) embryo culture-derived seedlings

Ex vitro rooting using a mini growth chamber to maintain relative humidity has been used to mass produce true-to-type Kopyor coconut (Cocos nucifera L.) seedlings through embryo culture. This new process was found to (1) improve the proportion of seedlings successfully transferred to soil (from 40 to 90% for seedlings with roots); (2) achieve this step in the shortest time possible (a reduction from 10 to 4 mo in in vitro culture); (3) improve root formation using indolebutyric acid (IBA; an improvement in the number of primary roots from 2 to 5); and (4) improve the vigor of seedlings ex vitro (improvements of fresh weight, shoot length, number of opened leaves, leaf thickness, amount of epicuticular wax, and stomatal density). The best ex vitro rooting and rapid acclimatization protocol was obtained when 4-mo-old seedlings with two opened leaves were kept in the mini growth chamber for 3 mo before being transferred into soil, and when the mini growth chamber was flooded with a quarter strength hybrid embryo culture (HEC) medium with 1 μM IBA but depleted of vitamins and sugar. This protocol was efficient in delivering high-value Kopyor seedlings to the field (90% success rate), with a decreased risk of contamination and lower labor cost. The improved process was found applicable to both tall and dwarf Kopyor and other coconut types.     

Elevated CO2 induces differences in nodulation of soybean depending on bradyrhizobial strain and method of inoculation

High CO₂ has been shown to increase plant growth and to affect symbiotic activity in many legumes species, including soybean (Glycine max [L.] Merr.). In order to assess the interaction between elevated CO₂ and rhizobial symbionts on soybean growth and nodulation, we combined the effects of CO₂ with those of different bradyrhizobial strains and methods of inoculation. Soybean seeds were sown in agricultural soil in pots and inoculated with three strains of Bradyrhizobium japonicum (5Sc2 and 12NS14 indigenous to Quebec soils, and 532c, a reference strain), the inoculum being either applied directly to the seed or incorporated into the soil. Plants were grown in growth chambers (22/17ºC) for 6 weeks, under either near ambient (400 μmol mol^?1) or elevated (800 μmol mol^?1) concentrations of CO_2. Elevated CO₂ increased mass (63%) and number (50%) of soybean nodules, particularly medium and large, allowed a deeper nodule development, and increased shoot dry weight (+30%), shoot C uptake (+33%) and shoot N uptake (+78%), compared to ambient CO₂. The two indigenous strains induced more medium and large nodules under elevated CO₂ than the reference strain and showed the greatest increases in shoot dry weight. Soil inoculation induced higher number of small nodules than seed inoculation, specifically for the two indigenous strains, but did not affect plant growth parameters. We conclude that soybean yield enhancements due to elevated CO₂ are associated with the production of large and medium-size nodules and a deep nodulation, that the two indigenous strains better respond to elevated CO₂ than the reference strain, and that the method of inoculation has little influence on this response.     

In vitro photoautotrophic acclimatization,direct transplantation and ex vitro adaptation of rubber tree (<Emphasis Type="Italic">Hevea brasiliensis</Emphasis>)

We investigated the effect of carbon dioxide (CO₂)-ambient (350 µmol CO₂ mol^?1) and CO₂-enriched (1500 µmol CO₂ mol^?1) conditions of in vitro photoautotrophic system on two cultivars, ‘RRIM600’ and ‘RRIT413’ of rubber tree (Hevea brasiliensis) in an acclimatization process of 45 days. Survival percentage of in vitro rubber tree plantlets derived from somatic embryos under ambient CO₂ was better than those under CO₂-enriched conditions, especially in cv. ‘RRIT413’. Subsequently, the survival rate of ex vitro transplanted plantlets was similar to the in vitro plantlets and abnormal morphological characters such as light-green leaves (SPAD), small leaves in cv. ‘RRIT413’ acclimatized under CO₂-enriched conditions were demonstrated 30 days after the plantlets were transferred into the soil. Maximum quantum yield of PSII, photon yield of PSII, stomatal conductance and transpiration rate in cv. ‘RRIT413’ acclimatized under CO₂-enriched conditions were sharply declined by 39.0, 50.6, 47.1 and 45.8%, respectively as compared to those acclimatized under ambient CO₂ conditions. In contrast, the in vitro acclimatized plantlets of cv. ‘RRIM600’ were un-responsive under both ambient- and enriched-CO₂ conditions. In conclusion, genotypic dependent in response to CO₂ enriched conditions in in-vitro acclimatization of rubber tree plantlets was evidently demonstrated as a key result to regulate plant growth and development in ex vitro environments. Interestingly, soluble sugar contents (sucrose, glucose and fructose) were increased after transplanting the plantlets of cv. ‘RRIM600’ acclimatized under CO₂-enriched condition into the soil and thus, can be considered as an adaptive indicator of ex vitro adaptation.     

Genotypes of Brassica rapa respond differently to plant-induced variation in air CO2 concentration in growth chambers with standard and enhanced venting

Growth chambers allow measurement of phenotypic differences among genotypes under controlled environment conditions. However, unintended variation in growth chamber air CO₂ concentration ([CO₂]) may affect the expression of diverse phenotypic traits, and genotypes may differ in their response to variation in [CO₂]. We monitored [CO₂] and quantified phenotypic responses of 22 Brassica rapa genotypes in growth chambers with either standard or enhanced venting. [CO₂] in chambers with standard venting dropped to 280 μmol mol^?1 during the period of maximum canopy development, ~80 μmol mol^?1 lower than in chambers with enhanced venting. The stable carbon isotope ratio of CO₂ in chamber air (δ¹³C_air) was negatively correlated with [CO₂], suggesting that photosynthesis caused observed [CO₂] decreases. Significant genotype × chamber-venting interactions were detected for 12 of 20 traits, likely due to differences in the extent to which [CO₂] changed in relation to genotypes’ phenology or differential sensitivity of genotypes to low [CO₂]. One trait, ¹³C discrimination (δ¹³C), was particularly influenced by unaccounted-for fluctuations in δ¹³C_air and [CO₂]. Observed responses to [CO₂] suggest that genetic variance components estimated in poorly vented growth chambers may be influenced by the expression of genes involved in CO₂ stress responses; genotypic values estimated in these chambers may likewise be misleading such that some mapped quantitative trait loci may regulate responses to CO₂ stress rather than a response to the environmental factor of interest. These results underscore the importance of monitoring, and where possible, controlling [CO₂].     

Functional characteristics of a fruticose type of lichen, Stereocaulon foliolosum Nyl. in response to light and water stress

Stereocaulon foliolosum a fruticose type of lichen under its natural habitat is subjected to low temperature, high light conditions and frequent moisture stress due its rocky substratum. To understand as to how this lichen copes up with these stresses, we studied the reflectance properties, light utilization capacity and the desiccation tolerance under laboratory conditions. S. foliolosum showed light saturation point for photosynthesis at 390 μmol CO₂ m^?2 s^?1 and the light compensation point for photosynthesis at 64 μmol CO₂ m^?2 s^?1. Our experiments show that S. foliolosum has a low absorptivity (30–35 %) towards the incident light. The maximum rates of net photosynthesis and apparent electron transport observed were 1.9 μmol CO₂ m^?2 s^?1 and 45 μmol e^? m^?2 s^?1, respectively. The lichen recovers immediately after photoinhibition under low light conditions. S. foliolosum on subjecting to desiccation results in the decrease of light absorptivity and the reflectance properties associated with water status of the thalli show a change. During desiccation, a simultaneous decrease in photosynthesis, dark respiration and quenching in the fluorescence properties was observed. However, all the observed changes show a rapid recovery on rewetting the lichen. Our study shows that desiccation does not have a severe or long-term impact on S. foliolosum and the lichen is also well adapted to confront high light intensities.     

Effects of Gamma Stress and Carbon Dioxide on Eight Bioactive Flavonoids and Photosynthetic Efficiency in <Emphasis Type="Italic">Centella asiatica</Emphasis>

Increased atmospheric CO₂ and gamma irradiation have a significant impact on the plant photosynthetic apparatus and organic compound production. In this study, we evaluated the effect of elevated CO₂ on the photosynthetic efficiency and production of defensive secondary metabolites (flavonoids) induced by gamma irradiation as a physical elicitor in Centella asiatica. Irradiated and non-irradiated 10-week-old plants of C. asiatica were exposed to 400 and 800 μmol mol^?1 of atmospheric CO₂ in growth chambers for 2 h every day until six weeks. A CO₂-enriched atmosphere initially improved the photosynthetic efficiency and ameliorated the detrimental impact of gamma irradiation on the photosynthetic apparatus, increasing carbon allocation into the flavonoid pathway. Elevated CO₂ combined with gamma irradiation resulted in the highest concentration of flavonoids in C. asiatica tissues compared with the other treatments. There was an enhancement in rutin (2.49 fold), naringin (2.15 fold), fisetin (4.07 fold), and morin (4.62 fold) with rising CO₂ concentrations from 400 to 800 μmol mol^?1 in the irradiated plants. With increasing CO₂ concentration, the compensation point and the respiration declined, whereas the apparent quantum yield and the maximum net photosynthesis (A _max) rate increased. The efficiency of photosystem II (PSII) was improved in the irradiated plants grown under high concentrations of CO₂. The total carbohydrate concentration reached the maximum value at the highest level of CO₂, followed by gamma irradiation combined with the highest level of CO₂. Irradiated plants of C. asiatica grown under elevated CO₂ could be superior to non-irradiated plants due to increased carbon availability both for the flavonoid biosynthesis and for the photosynthetic pathway.     

Ecosystem respiration derived from 222Rn measurements on Rishiri Island, Japan

We evaluated the nighttime CO₂ flux (ecosystem respiration) on Rishiri Island, located at the northern tip of Hokkaido, Japan, from 2009 to 2011, by using the relationship between atmospheric ²²²Rn and CO₂ concentrations. The annual mean CO₂ flux was 1.8 μmol m^?2 s^?1, with a maximum monthly mean in July (4.6 ± 2.6 μmol m^?2 s^?1) and a broad minimum from December to March (0.33 ± 0.29 μmol m^?2 s^?1). The annual mean was comparable to fluxes at the JapanFlux sites in northern Japan. During the season of snow cover (mid-December to early April), the CO₂ flux was low (0.45 ± 0.43 μmol m^?2 s^?1). Total annual respiration was estimated at 679 ± 174 g cm^?2, about 8 % of which occurred during the season of snow cover.     

Impact of elevated CO<Subscript>2</Subscript> on root traits of a sapling community of three birches and an oak: a free-air-CO<Subscript>2</Subscript> enrichment (FACE) in northern Japan

Key message

The CO ₂ effect on the root production of a broad-leaved community was insignificant when grown in brown forest soil, however, it was positively large when grown in volcanic ash soil.

Abstract

We evaluated the root response to elevated CO₂ fumigation of 3 birches (Betula sp.) and 1 deciduous oak (Quercus sp.) grown in immature volcanic ash soil (VA) or brown forest soil (BF). VA is a nutrient-poor, phosphorus-impoverished soil, broadly distributed in northern Japan. Each species had been exposed to either ambient (375–395 μmol mol^?1) (aCO₂) or elevated (500 μmol mol^?1) (eCO₂) CO₂ during the daytime (more than 70 μmol m^?2 s^?1) over 4 growing seasons. The results suggest that eCO₂ did not cause an increase in total root production when the community had grown in fertile BF soil, however, it did cause a large increase when the community was grown in infertile VA soil. Yet, carbon allocation to plant roots was not affected by eCO₂ in either the BF or VA soils. Rhizo-morphogenesis appeared to occur to a greater extent under eCO₂. It seems that the saplings developed a massive amount of fine roots under the VA and eCO₂ conditions. Unexpectedly, eCO₂ resulted in a larger total root mass when the community was grown in VA soil than when grown in BF soil (eCO₂ × VA vs. eCO₂ × BF). These results may hint to a site-specific potential of communities to sequester future atmospheric carbon. The growing substance of plants is an important factor which root response to eCO₂ depends on, however, further studies are needed for a better understanding.

     

In vitro and in vivo analyses of the role of the carboxysomal β-type carbonic anhydrase of the cyanobacterium Synechococcus elongatus in carboxylation of ribulose-1,5-bisphosphate

The carboxylase activities of crude carboxysome preparations obtained from the wild-type Synechococcus elongatus strain PCC 7942 strain and the mutant defective in the carboxysomal carbonic anhydrase (CA) were compared. The carboxylation reaction required high concentrations of bicarbonate and was not even saturated at 50 mM bicarbonate. With the initial concentrations of 50 mM and 25 mM for bicarbonate and ribulose-1,5-bisphosphate (RuBP), respectively, the initial rate of RuBP carboxylation by the mutant carboxysome (0.22 μmol mg^?1 protein min^?1) was only 30 % of that observed for the wild-type carboxysomes (0.71 μmol mg^?1 protein min^?1), indicating the importance of the presence of CA in efficient catalysis by ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). While the mutant defective in the ccmLMNO genes, which lacks the carboxysome structure, could grow under aeration with 2 % (v/v) CO₂ in air, the mutant defective in ccaA as well as ccmLMNO required 5 % (v/v) CO₂ for growth, indicating that the cytoplasmically localized CcaA helped utilization of CO₂ by the cytoplasmically localized Rubisco by counteracting the action of the CO₂ hydration mechanism. The results predict that overexpression of Rubisco would hardly enhance CO₂ fixation by the cyanobacterium at CO₂ levels lower than 5 %, unless Rubisco is properly organized into carboxysomes.     

Elevated CO2 enhances the growth of hybrid larch F1 (Larix gmelinii var. japonica × L. kaempferi) seedlings and changes its biomass allocation

Key message

Elevated CO ₂ enhances the photosynthesis and growth of hybrid larch F ₁ seedlings. However, elevated CO ₂ -induced change of tree shape may have risk to the other environmental stresses.

Abstract

The hybrid larch F₁ (Larix gmelinii var. japonica × L. kaempferi) is one of the most promising species for timber production as well as absorption of atmospheric CO₂. To assess the ability of this species in the future high CO₂ environment, we investigated the growth and photosynthetic response of hybrid larch F₁ seedlings to elevated CO₂ concentration. Three-year-old seedlings of hybrid larch F₁ were grown on fertile brown forest soil or infertile volcanic ash soil, and exposed to 500 μmol mol^?1 CO₂ in a free-air CO₂ enrichment system located in northern Japan for two growing seasons. Regardless of soil type, the exposure to elevated CO₂ did not affect photosynthetic traits in the first and second growing seasons; a higher net photosynthetic rate was maintained under elevated CO₂. Growth of the seedlings under elevated CO₂ was greater than that under ambient CO₂. We found that elevated CO₂ induced a change in the shape of seedlings: small roots, slender-shaped stems and long-shoots. These results suggest that elevated CO₂ stimulates the growth of hybrid larch F₁, although the change in tree shape may increase the risk of other stresses, such as strong winds, heavy snow, and nutrient deficiency.     

Fed-Batch Cultivation of Arthrospira platensis Using Carbon Dioxide from Alcoholic Fermentation and Urea as Carbon and Nitrogen Sources

To reduce CO₂ emissions from alcoholic fermentation, Arthrospira platensis was cultivated in tubular photobioreactor using either urea or nitrate as nitrogen sources at different light intensities (60 μmol m^?2 s^?1?≤?I?≤?240 μmol m^?2 s^?1). The type of carbon source (pure CO₂ or CO₂ from fermentation) did not show any appreciable influence on the main cultivation parameters, whereas substitution of nitrate for urea increased the nitrogen-to-cell conversion factor (Y _X/N ), and the maximum cell concentration (X _m ) and productivity (P _X ) increased with I. As a result, the best performance using gaseous emissions from alcoholic fermentation (X _m ?=?2,960?±?35 g m^?3, P _X ?=?425?±?5.9 g m^?3 day^?1 and Y _X/N ?=?15?±?0.2 g g^?1) was obtained at I?=?120 μmol m^?2 s^?1 using urea as nitrogen source. The results obtained in this work demonstrate that the combined use of effluents rich in urea and carbon dioxide could be exploited in large-scale cyanobacteria cultivations to reduce not only the production costs of these photosynthetic microorganisms but also the environmental impact associated to the release of greenhouse emissions.     

Micropropagation of <Emphasis Type="Italic">Glossonema varians</Emphasis> (Stocks) Benth. ex Hook.f.—a rare Asclepiadeae of Indian Thar Desert

In the present study, an in vitro regeneration protocol for Glossonema varians (Stocks) Benth. ex Hook.f. of family Asclepiadaceae was optimized. Cotyledonary nodes of in vitro cultured seedlings were used as explants for activation of axillary shoot buds. Axillary shoot buds were initially activated on 0.1 mg L^?1 6-benzylaminopurine (BAP) and then multiplied on 0.05 mg L^?1 BAP. Shoots were rooted in vitro on 1/4 strength Murashige and Skoog medium containing 0.1 mg L^?1 2-naphthoxyacetic acid and 100 mg L^?1 activated charcoal. The cultures were maintained in a 12 h photoperiod at 40–50 μmol m^?2 s^?1 spectral flux photon, 25–30?±?2°C, and 60% relative humidity (RH). Up to 80% of in vitro regenerated plantlets were acclimatized on soilrite in cotton-plugged culture tubes in the greenhouse. This protocol can be a useful method to mass propagate and conserve this rare plant to balance biodiversity in the desert ecosystem.     

Meta-analysis of the interaction between shade-tolerance,light environment and growth response of woody species to elevated CO2

Growth responses of different tree species to elevated CO₂ vary enormously, even when grown under the same basic experimental conditions (such as nutrient supply and light conditions). A test of the hypothesis that this variation is associated with the species’ shade-tolerance is presented. A meta-analysis of 74 logarithm-transformed response ratios RR (total biomass in ∼700 μmol·mol^–1 CO₂/biomass in current ambient CO₂), grouped according to the species’ relative tolerance to shade, revealed that RR differed widely and significantly between seedlings or saplings of woody species of different shade-tolerance. Only comparative studies were included in the meta-analysis, and interference from differences in basic experimental conditions between experiments was minimised. The mean RR of species with very high shade-tolerance was 1.51. This was significantly greater than the value of 1.18 for species intolerant of shade. Smaller differences in mean RR were found between groups of species that differed less widely in shade-tolerance. When responses were categorised according to two aspects of light environment in the experiments, daily integrated or peak photosynthetically active photon flux density (PFD), the differences in RR between different shade-tolerance classes disappeared at daily integrated PFD below ∼20 mol·m^–2·d^–1 or peak PFD below ∼250 μmol·m^–2·s^–1. Growth stimulation in juvenile trees is likely to cause increased survivorship in subsequent years. Even if the observed differences in RR were restricted to sites experiencing no or only moderate shading, it appears likely that they will affect species composition of naturally regenerating forests in the long term.     

Photosynthetic capacity of the capsule wall and its contribution to carbon fixation and seed yield in castor (<Emphasis Type="Italic">Ricinus communis</Emphasis> L.)

Defoliation occurs in castor due to several reasons, but the crop has propensity to compensate for the seed yield. Photosynthetic efficiency in terms of functional (gas exchange and chlorophyll fluorescence) and structural characteristics (photosynthetic pigment profiles and anatomical properties) of castor capsule walls under light- and dark-adapted conditions was compared with that of leaves. Capsule wall showed high intrinsic efficiency of photosystem II (F _v/F _m, 0.82) which was comparable to leaves (F _v/F _m, 0.80). With increasing photon flux densities (PFD), actual quantum yields and photochemical quenching coefficients of the capsule walls were similar to that in leaves, while electron transport rates reached a maximum corresponding to about 118 % of the leaves. However, maximum net photosynthetic rate of the capsule walls (2.60 µmol CO₂ m^?2 s^?1) was less than one-fourth of the leaves (15.67 µmol CO₂ m^?2 s^?1) at the CO₂ concentration of 400 µmol mol^?1, and the difference was attributed to about 80 % lower stomatal density and the 75 % lower total chlorophyll content of capsule walls than the leaves. Furthermore, seed weight in dark-adapted capsules was 2.70–12.42 % less as compared to the capsules developed under light. The results indicate that castor capsule walls are photosynthetically active (about 15–30 % of the leaves) and contribute significantly to carbon fixation and seed yield accounting for 10 % photoassimilates towards seed weight.     

Short-term impacts of active layer detachments on carbon exchange in a High Arctic ecosystem,Cape Bounty,Nunavut, Canada

Localized permafrost disturbances such as active layer detachments (ALDs) are increasing in frequency and severity across the Canadian Arctic impacting terrestrial ecosystem functioning. However, the contribution of permafrost disturbance-carbon feedbacks to the carbon (C) balance of Arctic ecosystems is poorly understood. Here, we explore the short-term impact of active layer detachments (ALDs) on carbon dioxide (CO₂) exchange in a High Arctic semi-desert ecosystem by comparing midday C exchange between undisturbed areas, moderately disturbed areas (intact islands of vegetation within an ALD), and highly disturbed areas (non-vegetated areas due to ALD). Midday C exchange was measured using a static chamber method between June 23 and August 8 during the 2009 and 2010 growing seasons. Results show that areas of high disturbance had significantly reduced gross ecosystem exchange and ecosystem respiration (R _E) compared to control and moderately disturbed areas. Moderately disturbed areas showed significantly enhanced net ecosystem exchange compared to areas of high disturbance, but were not significantly different from control areas. Disturbance did not significantly impact soil thermal, physical or chemical properties. According to average midday fluxes, ALDs as a whole (moderately disturbed areas: ?1.942 μmol m^?2 s^?1+ highly disturbed areas: 2.969 μmol m^?2 s^?1) were a small CO₂ source of 1.027 μmol m^?2 s^?1 which did not differ significantly from average midday fluxes in control areas 1.219 μmol m^?2 s^?1. The findings of this study provide evidence that the short-term impacts of ALDs on midday, net C exchange and soil properties in a High Arctic semi-desert are minimal.     

Biochemical and biophysical CO2 concentrating mechanisms in two species of freshwater macrophyte within the genus Ottelia (Hydrocharitaceae)

Two freshwater macrophytes, Ottelia alismoides and O. acuminata, were grown at low (mean 5 μmol L^?1) and high (mean 400 μmol L^?1) CO₂ concentrations under natural conditions. The ratio of PEPC to RuBisCO activity was 1.8 in O. acuminata in both treatments. In O. alismoides, this ratio was 2.8 and 5.9 when grown at high and low CO₂, respectively, as a result of a twofold increase in PEPC activity. The activity of PPDK was similar to, and changed with, PEPC (1.9-fold change). The activity of the decarboxylating NADP-malic enzyme (ME) was very low in both species, while NAD-ME activity was high and increased with PEPC activity in O. alismoides. These results suggest that O. alismoides might perform a type of C₄ metabolism with NAD-ME decarboxylation, despite lacking Kranz anatomy. The C₄-activity was still present at high CO₂ suggesting that it could be constitutive. O. alismoides at low CO₂ showed diel acidity variation of up to 34 μequiv g^?1 FW indicating that it may also operate a form of crassulacean acid metabolism (CAM). pH-drift experiments showed that both species were able to use bicarbonate. In O. acuminata, the kinetics of carbon uptake were altered by CO₂ growth conditions, unlike in O. alismoides. Thus, the two species appear to regulate their carbon concentrating mechanisms differently in response to changing CO₂. O. alismoides is potentially using three different concentrating mechanisms. The Hydrocharitaceae have many species with evidence for C₄, CAM or some other metabolism involving organic acids, and are worthy of further study.     

Nutrient uptake rates by the alien alga Undaria pinnatifida (Phaeophyta) (Nuevo Gulf, Patagonia, Argentina) when exposed to diluted sewage effluent

In the early nineties, Undaria pinnatifida has been accidentally introduced to Nuevo Gulf (Patagonia, Argentina) where the environmental conditions would have favored its expansion. The effect of the secondary treated sewage discharge from Puerto Madryn city into Nueva Bay (located in the western extreme of Nuevo Gulf) is one of the probable factors to be taken into account. Laboratory cultures of this macroalgae were conducted in seawater enriched with the effluent. The nutrients (ammonium, nitrate and phosphate) uptake kinetics was studied at constant temperature and radiation (16?°C and 50 μE m^?2 s^?1 respectively). Uptake kinetics of both inorganic forms of nitrogen were described by the Michaelis–Menten model during the surge phase (ammonium: V _max ^sur: 218.1 μmol h^?1 g^?1, K _s ^sur: 476.5 μM and nitrate V _max ^sur: 10.7 μmol h^?1 g^?1, K _s ^sur: 6.1 μM) and during the assimilation phase (ammonium: V _max ^ass: 135.6 μmol h^?1 g^?1, K _s ^ass: 407.2 μM and nitrate V _max ^ass: 1.9 μmol h^?1 g^?1, K _s ^ass: 2.2 μM), with ammonium rates always higher than those of nitrate. Even though a net phosphate disappearance was observed in all treatments, uptake kinetics of this ion could not be properly estimated by the employed methodology.