Rubisco activase constrains the photosynthetic potential of leaves at high temperature and CO2

Net photosynthesis (Pn) is inhibited by moderate heat stress. To elucidate the mechanism of inhibition, we examined the effects of temperature on gas exchange and ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) activation in cotton and tobacco leaves and compared the responses to those of the isolated enzymes. Depending on the CO2 concentration, Pn decreased when temperatures exceeded 35–40°C. This response was inconsistent with the response predicted from the properties of fully activated Rubisco. Rubisco deactivated in leaves when temperature was increased and also in response to high CO2 or low O2. The decrease in Rubisco activation occurred when leaf temperatures exceeded 35°C, whereas the activities of isolated activase and Rubisco were highest at 42°C and >50°C, respectively. In the absence of activase, isolated Rubisco deactivated under catalytic conditions and the rate of deactivation increased with temperature but not with CO2. The ability of activase to maintain or promote Rubisco activation in vitro also decreased with temperature but was not affected by CO2. Increasing the activase/Rubisco ratio reduced Rubisco deactivation at higher temperatures. The results indicate that, as temperature increases, the rate of Rubisco deactivation exceeds the capacity of activase to promote activation. The decrease in Rubisco activation that occurred in leaves at high CO2 was not caused by a faster rate of deactivation, but by reduced activase activity possibly in response to unfavorable ATP/ADP ratios. When adjustments were made for changes in activation state, the kinetic properties of Rubisco predicted the response of Pn at high temperature and CO2.

Regulation of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase by Carbamylation and 2-Carboxyarabinitol 1-Phosphate in Tobacco: Insights from Studies of Antisense Plants Containing Reduced Amounts of Rubisco Activase1

The regulation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity by 2-carboxyarabinitol 1-phosphate (CA1P) was investigated using gas-exchange analysis of antisense tobacco (Nicotiana tabacum) plants containing reduced levels of Rubisco activase. When an increase in light flux from darkness to 1200 μmol quanta m−2 s−1 was followed, the slow increase in CO2 assimilation by antisense leaves contained two phases: one represented the activation of the noncarbamylated form of Rubisco, which was described previously, and the other represented the activation of the CA1P-inhibited form of Rubisco. We present evidence supporting this conclusion, including the observation that this second phase, like CA1P, is only present following darkness or very low light flux. In addition, the second phase of CO2 assimilation was correlated with leaf CA1P content. When this novel phase was resolved from the CO2 assimilation trace, most of it was found to have kinetics similar to the activation of the noncarbamylated form of Rubisco. Additionally, kinetics of the novel phase indicated that the activation of the CA1P-inhibited form of Rubisco proceeds faster than the degradation of CA1P by CA1P phosphatase. These results may be significant with respect to current models of the regulation of Rubisco activity by Rubisco activase.

Physiological responses to water stress in Eucalyptus cloeziana and E-argophloia seedlings

Effects of water stress duration and intensity on gas exchange and leaf water potential were investigated in 7-month-old seedlings of a humid coastal provenance (Gympie) and a dry inland (Hungry Hills) provenance of E. cloeziana F. Muell. and in a dry inland (Chinchilla) provenance of E. argophloia Blakely supplied with 100% (T-100), 70% (T-70), 50% (T-50) of their water requirements, or were watered only after they were wilted at dawn (T-0). Seedlings of E. argophloia had the highest midday net photosynthetic rate (A), stomata] conductance (g(s)), stomatal density and predawn leaf water potential (Psi(pd)) in all treatments. The E. cloeziana provenances did not differ in these attributes. The T-70 and T-50 treatments caused reductions in A of 30% in E. argophloia, and 55% in the E. cloeziana provenances. Under the T-0 treatment, E. argophloia maintained higher rates of gas exchange at all levels of water stress than E. cloeziana provenances. The estimates of Psi(pd) and midday water potential (Psi(md)) at which plants remained wilted overnight were respectively: -2.7 and -4.1 MPa for E. cloeziana (humid), -2.8 and -4.0 MPa for E. cloeziana (dry) and, -3.7 and -4.9 MPa for E. argophloia. Following stress relief, both A and g(s) recovered more quickly in E. argophloia and in the dry provenance of E. cloeziana than in the humid provenance. We conclude that E. argophloia is more drought tolerant and has a potential for cultivation in the humid and semi humid climates, whilst E. cloeziana has greater potential in the humid subtropical climates.

Differences in plant function in phosphorus- and nitrogen-limited mangrove ecosystems

Mangrove ecosystems can be either nitrogen (N) or phosphorus (P) limited and are therefore vulnerable to nutrient pollution. Nutrient enrichment with either N or P may have differing effects on ecosystems because of underlying differences in plant physiological responses to these nutrients in either N- or P-limited settings. Using a common mangrove species, Avicennia germinans, in sites where growth was either N or P limited, we investigated differing physiological responses to N and P limitation and fertilization. We tested the hypothesis that water uptake and transport, and hydraulic architecture, were the main processes limiting productivity at the P-limited site, but that this was not the case at the N-limited site. We found that plants at the P-deficient site had lower leaf water potential, stomatal conductance and photosynthetic carbon-assimilation rates, and less conductive xylem, than those at the N-limited site. These differences were greatly reduced with P fertilization at the P-limited site. By contrast, fertilization with N at the N-limited site had little effect on either photosynthetic or hydraulic traits. We conclude that growth in N- and P-limited sites differentially affect the hydraulic pathways of mangroves. Plants experiencing P limitation appear to be water deficient and undergo more pronounced changes in structure and function with relief of nutrient deficiency than those in N-limited ecosystems.

Respostas ecofisiológicas de Cybistax antisyphilitica Mart. (Ipê verde) em função das alterações na intensidade de luz

Light varies widely in both time and space in forest formation of “Bioma Cerrado”. Cybistax antisyphilitica occurs in areas typical of this biome, such as cerrado sensu stricto, “cerradões”, and altered areas. The aim of this study was to understand the morphological and physiological responses of C. antisyphilitica to alterations in light intensity. Juvenile plants (5 month of age) were taken to a fragment of semideciduous forest in Uberlândia-MG, and were divided into three treatments: 50 were maintained under the canopy (UC) 20 were kept in small gap (SG) and 20 were maintained under in full sun (FS). The daily courses of chlorophyll a fluorescence were made at the beginning, middle and end of dry season in 2015. At the end of the experiment measurements of chlorophyll content, gas exchange and growth were made. The plants showed dynamic photoinhibition as exhibited by reductions on Fv/Fm close to midday at the end of the dry season. Regarding the effective quantum yield (ΔF/Fm'), plants under FS showed reduced values that coincided with the higher values of electron transport rates (ETR). Plants under FS showed higher values of net CO2 assimilation rates, stomatal conductance, transpiration rates, water use efficiency and chlorophyll content compared to plants under UC. The stem diameter, dry mass of leaves and stem, total dry mass and relative growth rate were higher in plants under FS than plants under UC. On the other hand, plants under UC showed superior values of height, specific leaf area and leaf area ratio. Our results indicate that C. antisyphilitica has plasticity to survive in the contrasting light environments of the semideciduous forests, but this species was able to growth better under full sun conditions.

The organic sea-surface microlayer in the upwelling region off the coast of Peru and potential implications for air-sea exchange processes

The sea-surface microlayer (SML) is at the upper- most surface of the ocean, linking the hydrosphere with the atmosphere. The presence and enrichment of organic compounds in the SML have been suggested to influence air- sea gas exchange processes as well as the emission of primary organic aerosols. Here, we report on organic matter components collected from an approximately 50µm thick SML and from the underlying water (ULW), ca. 20 cm below the SML, in December 2012 during the SOPRAN METEOR 91 cruise to the highly productive, coastal upwelling regime off the coast of Peru. Samples were collected at 37 stations including coastal upwelling sites and off-shore stations with less organic matter and were analyzed for total and dissolved high molecular weight (> 1 kDa) combined carbohydrates (TCCHO, DCCHO), free amino acids (FAA), total and dissolved hydrolyzable amino acids (THAA, DHAA), transparent exopolymer particles (TEP), Coomassie stainable particles (CSPs), total and dissolved organic carbon (TOC, DOC), total and dissolved nitrogen (TN, TDN), as well as bacterial and phytoplankton abundance. Our results showed a close coupling between organic matter concentrations in the water column and in the SML for almost all components except for FAA and DHAA that showed highest enrichment in the SML on average. Accumulation of gel particles (i.e., TEP and CSP) in the SML differed spatially. While CSP abundance in the SML was not related to wind speed, TEP abundance decreased with wind speed, leading to a depletion of TEP in the SML at about 5 m s-1 . Our study provides insight to the physical and biological control of organic matter enrichment in the SML, and discusses the potential role of organic matter in the SML for air-sea exchange processes.

Air-sea transfer of gas phase controlled compounds

Gases in the atmosphere/ocean have solubility that spans several orders of magnitude. Resistance in the molecular sublayer on the waterside limits the air-sea exchange of sparingly soluble gases such as SF6 and CO2. In contrast, both aerodynamic and molecular diffusive resistances on the airside limit the exchange of highly soluble gases (as well as heat). Here we present direct measurements of air-sea methanol and acetone transfer from two open cruises: the Atlantic Meridional Transect in 2012 and the High Wind Gas Exchange Study in 2013. The transfer of the highly soluble methanol is essentially completely airside controlled, while the less soluble acetone is subject to both airside and waterside resistances. Both compounds were measured concurrently using a proton-transfer-reaction mass spectrometer, with their fluxes quantified by the eddy covariance method. Up to a wind speed of 15 m s-1, observed air-sea transfer velocities of these two gases are largely consistent with the expected near linear wind speed dependence. Measured acetone transfer velocity is ~30% lower than that of methanol, which is primarily due to the lower solubility of acetone. From this difference we estimate the "zero bubble" waterside transfer velocity, which agrees fairly well with interfacial gas transfer velocities predicted by the COARE model. At wind speeds above 15 m s-1, the transfer velocities of both compounds are lower than expected in the mean. Air-sea transfer of sensible heat (also airside controlled) also appears to be reduced at wind speeds over 20 m s-1. During these conditions, large waves and abundant whitecaps generate large amounts of sea spray, which is predicted to alter heat transfer and could also affect the air-sea exchange of soluble trace gases. We make an order of magnitude estimate for the impacts of sea spray on air-sea methanol transfer.

Air-sea transfer of gas phase controlled compounds

Gases in the atmosphere/ocean have solubility that spans several orders of magnitude. Resistance in the molecular sublayer on the waterside limits the air-sea exchange of sparingly soluble gases such as SF6 and CO2. In contrast, both aerodynamic and molecular diffusive resistances on the airside limit the exchange of highly soluble gases (as well as heat). Here we present direct measurements of air-sea methanol and acetone transfer from two open cruises: the Atlantic Meridional Transect in 2012 and the High Wind Gas Exchange Study in 2013. The transfer of the highly soluble methanol is essentially completely airside controlled, while the less soluble acetone is subject to both airside and waterside resistances. Both compounds were measured concurrently using a proton-transfer-reaction mass spectrometer, with their fluxes quantified by the eddy covariance method. Up to a wind speed of 15 m s-1, observed air-sea transfer velocities of these two gases are largely consistent with the expected near linear wind speed dependence. Measured acetone transfer velocity is ~30% lower than that of methanol, which is primarily due to the lower solubility of acetone. From this difference we estimate the "zero bubble" waterside transfer velocity, which agrees fairly well with interfacial gas transfer velocities predicted by the COARE model. At wind speeds above 15 m s-1, the transfer velocities of both compounds are lower than expected in the mean. Air-sea transfer of sensible heat (also airside controlled) also appears to be reduced at wind speeds over 20 m s-1. During these conditions, large waves and abundant whitecaps generate large amounts of sea spray, which is predicted to alter heat transfer and could also affect the air-sea exchange of soluble trace gases. We make an order of magnitude estimate for the impacts of sea spray on air-sea methanol transfer.

IMPACTS OF ENHANCED TEMPERATURE AND SNOW DEPOSITION ON SENESCENCE DATE, VEGETATION COVER, AND CO2 EXCHANGE IN A CANADIAN HIGH ARCTIC MESIC ECOSYSTEM

Arctic regions are expected to experience an increase in both temperature and precipitation over the coming decades, which is likely to impact vegetation dynamics and greenhouse gas exchange. To test this response, an experiment was installed at the Cape Bounty Arctic Watershed Observatory, on Melville Island, NU, in 2008 as part of the International Tundra Experiment (ITEX). Snow fences and open top chambers (OTCs) were used to manipulate snow depth and air temperature, respectively. Unlike most ITEX sites to date, enhanced temperature and snowfall were combined here in a factorial design with eight replicates. As an added control, four plots were established well outside the enhanced snow area. Senescence date was recorded at the end of the season, and at the peak of the growing season a vegetation survey was conducted within each plot in order to determine the total percent cover of each plot, as well as the percent cover of individual species. Carbon dioxide (CO2) exchange was also measured within each plot throughout the growing season. The date of senescence occurred significantly earlier in plots which had not been manipulated in any way, compared to all other treatments for all species. Salix arctica showed the greatest increase in cover over time at the species level. Lichen cover increased significantly in the deepened snow plots, and in general there were significant increases in percent cover in some functional groups over time. During June and into July the net CO2 flux was to the atmosphere. It was not until July 27 that these ecosystems became net carbon sinks. However, warming alone resulted in the ecosystem acting as a significant net carbon sink for the entire growing season. Plots exposed to warming alone were estimated to have removed approximately 19.94 g C m-2 from the atmosphere, whereas all other treatments were very similar to one another and estimated to have added approximately 3.12 g C m-2 to the atmosphere. Active layer depth and soil temperatures suggest that plots within the ambient snow zone may be receiving some additional snow due to their proximity to the fences. CO2 fluxes measured within the outer control plots suggest that the effect of warming alone could lead to this ecosystem being an even stronger net C sink under truly ambient snow conditions.

Linking thermal imaging to physiological indicators in Carica papaya L. under different watering regimes

Water deficit is the most limiting factor for yield and fruit-quality parameters in papaya crop (Carica papaya L.), deficit-irrigation (DI) strategies offering a feasible alternative to manage limiting water resources. When DI is applied, it is crucial to assess the physiological status of the crop in order to maintain the plant within a threshold value of water stress so as no to affect yield or fruit-quality parameters. The aim of this work was to evaluate the feasibility of thermal imaging in young papaya plants to assess the physiological status of this crop when it is subjected to different DI regimes, studying the relationships between the changes in leaf temperature (Tleaf) and in the major physiological parameters (i.e., stomatal conductance to water vapor, gs; transpiration, E; and net photosynthesis, An). The trial was conducted in a greenhouse from March to April of 2012. Plants were grown in pots and subjected to four irrigation treatments: (1) a full irrigation treatment (control), maintained at field capacity; (2) a partial root-zone drying treatment, irrigated with 50% of the total water applied to control to only one side of roots, alternating the sides every 7 days; (3) a regulated deficit irrigation (50% of the control, applied to both sides of plant); (4) and a non-irrigated treatment, in which irrigation was withheld from both sides of the split root for 14 days, followed by full irrigation until the end of the study. Significant relationships were found between Tleaf and major physiological variables such as gs, E and An. Additionally, significant relationships were found between the difference of leaf-to-air temperature (ΔTleaf–air) and gas-exchange measurements, which were used to establish the optimum range of ΔTleaf–air as a preliminary step to the crop-water monitoring and irrigation scheduling in papaya, using thermal imaging as the main source of information. According to the results, we conclude that thermal imaging is a promising technique to monitor the physiological status of papaya during drought conditions.

Linking thermal imaging to physiological indicators in Carica papaya L. under different watering regimes

Water deficit is the most limiting factor for yield and fruit-quality parameters in papaya crop (Carica papaya L.), deficit-irrigation (DI) strategies offering a feasible alternative to manage limiting water resources. When DI is applied, it is crucial to assess the physiological status of the crop in order to maintain the plant within a threshold value of water stress so as no to affect yield or fruit-quality parameters. The aim of this work was to evaluate the feasibility of thermal imaging in young papaya plants to assess the physiological status of this crop when it is subjected to different DI regimes, studying the relationships between the changes in leaf temperature (Tleaf) and in the major physiological parameters (i.e., stomatal conductance to water vapor, gs; transpiration, E; and net photosynthesis, An). The trial was conducted in a greenhouse from March to April of 2012. Plants were grown in pots and subjected to four irrigation treatments: (1) a full irrigation treatment (control), maintained at field capacity; (2) a partial root-zone drying treatment, irrigated with 50% of the total water applied to control to only one side of roots, alternating the sides every 7 days; (3) a regulated deficit irrigation (50% of the control, applied to both sides of plant); (4) and a non-irrigated treatment, in which irrigation was withheld from both sides of the split root for 14 days, followed by full irrigation until the end of the study. Significant relationships were found between Tleaf and major physiological variables such as gs, E and An. Additionally, significant relationships were found between the difference of leaf-to-air temperature (ΔTleaf–air) and gas-exchange measurements, which were used to establish the optimum range of ΔTleaf–air as a preliminary step to the crop-water monitoring and irrigation scheduling in papaya, using thermal imaging as the main source of information. According to the results, we conclude that thermal imaging is a promising technique to monitor the physiological status of papaya during drought conditions.

Nodulation, gas exchanges and production of peanut cultivated with Bradyrhizobium in soils with different textures.

Nitrogen fertilization from biological source is an uncommon practice for peanut growers due to the limited results, mainly in environments with water restriction. In this study, the response of a commercial Bradyrhizobium was evaluated on the nodulation and production of peanuts grown in sandy and medium textured soils. Two experiments using different soils were carried out in the field during the dry season, in Campina Grande, Paraíba State, Brazil. Three peanut genotypes were submitted to the following treatments: 1-no nitrogen fertilization (control), 2- chemical fertilization (ammonium sulfate) and 3- inoculation with Bradyrhizobium [commercial strain BR 1405 (SEMIA 6144)]. A completely randomized 3x3 factorial design was adopted with five repetitions for both experiments. The evaluates variables were: height of the main stem, number of nodes/plant, root length, root dry weight, weight of pods/plant and number of pods/plant. In addition, gas exchanges were estimated using IRGA apparatus. Both genotypes (BRS Havana and L7 Bege) were benefited in relation to production due to an inoculation with SEMIA 6144. No physiological response was verified in genotypes or N-treatments to gas exchange, excepting for the Ci/Ca ratio in the medium textured soil experiment. BRS Havana showed low Ci/Ca ratio in Bradyrhizobium treatment, indicating that SEMIA 6144 improved the plants photosynthetic efficiency.

Cultivation medium design via elemental balancing for tetraselmis suecica

The primary requirements for high-biomass-concentration microalgal cultivation include a photon source and distribution, efficient gas exchange and suitable growth medium composition. However, for mass outdoor production of microalgae, growth medium composition is a major controlling factor as most of the other factors such as light source and distribution are virtually uncontrollable. This work utilises an elemental balance approach between growth medium and biomass compositions to obtain high-density microalgal cultures in an open system. F medium, commonly used for the cultivation of marine microalgae such as Tetraselmis suecica was redesigned on the basis of increasing the biomass capacity of its major deficient components to support high biomass concentrations (τ ∼ 5.0 % for N, S and τ ∼ 10 % P), and the entire formulation was dissolved in 0.2 um sterile filtered natural seawater. Results show that the new medium (F') displayed a maximum biomass concentration and total lipid concentration of 1.29 g L 1 and 108.7 mg L 1 respectively, which represents over 2-fold increase compared to that of the F medium. Keeping all variables constant except growth medium, and using F medium as the base case of 1 medium cost (MC) unit mg -1 lipid, the F' medium yielded lipid at a cost of only 0.35 MC unit mg -1 lipids. These results show that greater amounts of biomass and lipids can be obtained more economically with minimal extra effort simply by using an optimised growth medium.

Packed bed bioreactor for the isolation and expansion of placental-derived Mesenchymal Stromal Cells

Large numbers of Mesenchymal stem/stromal cells (MSCs) are required for clinical relevant doses to treat a number of diseases. To economically manufacture these MSCs, an automated bioreactor system will be required. Herein we describe the development of a scalable closed-system, packed bed bioreactor suitable for large-scale MSCs expansion. The packed bed was formed from fused polystyrene pellets that were air plasma treated to endow them with a surface chemistry similar to traditional tissue culture plastic. The packed bed was encased within a gas permeable shell to decouple the medium nutrient supply and gas exchange. This enabled a significant reduction in medium flow rates, thus reducing shear and even facilitating single pass medium exchange. The system was optimised in a small-scale bioreactor format (160 cm2) with murine-derived green fluorescent protein-expressing MSCs, and then scaled-up to a 2800 cm2 format. We demonstrated that placental derived MSCs could be isolated directly within the bioreactor and subsequently expanded. Our results demonstrate that the closed system large-scale packed bed bioreactor is an effective and scalable tool for large-scale isolation and expansion of MSCs.

Genetic Diversity and Adaptation of Date Palm (Phoenix dactylifera L.)

Acquiring sufficient information on the genetic variation, genetic differentiation, and the ecological and genetic relationships among individuals and populations are essential for establishing guidelines on conservation and utilization of the genetic resources of a species, and more particularly when biotic and abiotic stresses are considered. The aim of this study was to assess the extent and pattern of genetic variation in date palm (Phoenix dacttylifera L) cultivars; the genetic diversity and structure in its populations occurring over geographical ranges; the variation in economically and botanically important traits of it and the variation in its drought adaptive traits, in conservation and utilization context. In this study, the genetic diversity and relationships among selected cultivars from Sudan and Morocco were assessed using microsatellite markers. Microsatellite markers were also used to investigate the genetic diversity within and among populations collected from different geographic locations in Sudan. In a separate investigation, fruits of cultivars selected from Sudan, involved morphological and chemical characterization, and morphological and DNA polymorphism of the mother trees were also investigated. Morphological and photosynthetic adjustments to water stress were studied in the five most important date palm cultivars in Sudan, namely, Gondaila, Barakawi, Bitamoda, Khateeb and Laggai; and the mechanism enhancing photosynthetic gas exchange in date palm under water stress was also investigated. Results showed a significant (p < 0.001, t-test) differentiation between Sudan and Morocco groups of cultivars. However, the major feature of all tested cultivars was the complete lack of clustering and the absence of cultivars representing specific clones. The results indicated high genetic as well as compositional and morphological diversity among cultivars; while, compositional and morphological traits were found to be characteristic features that strongly differentiate cultivars as well as phenotypes. High genetic diversity was observed also in different populations. Slight but significant (p < 0.01, AMOVA) divergence was observed for soft and dry types; however, the genetic divergence among populations was relatively weak. The results showed a complex genetic relationships between some of the tested populations especially when isolation by distance was considered. The results of the study also revealed that date palm cultivars and phenotypes possess specific direct or interaction effects due to water availability on a range of morphological and physiological traits. Soft and dry phenotypes responded differently to different levels of water stress, while the dry phenotype was more sensitive and conservative. The results indicated that date palm has high fixation capacity to photosynthetic CO2 supply with interaction effect to water availability, which can be considered as advantageous when coping with stresses that may arise with climate change. In conclusion, although a large amount of diversity exists among date palm germplasm, the findings in this study show that the role of biological nature of the tree, isolation by distance and environmental effects on structuring date palm genome was highly influenced by human impacts. Identity of date palm cultivars as developed and manipulated by date palm growers, in the absence of scientific breeding programmes, may continue to mainly depend on tree morphology and fruit characters. The pattern of genetic differentiation may cover specific morphological and physiological traits that contribute to adaptive mechanisms in each phenotype. These traits can be considered for further studies related to drought adaptation in date palm.