Andrea Scartazza

Comparisons of δ13C of photosynthetic products and ecosystem respiratory CO2 and their responses to seasonal climate variability

This study investigated the relationship between δ13C of ecosystem components, soluble plant carbohydrates and the isotopic signature of ecosystem respired CO2 (δ13CR) during seasonal changes in soil and atmospheric moisture in a beech (Fagus sylvatica L.) forest in the central Apennine mountains, Italy. Decrease in soil moisture and increase in air vapour pressure deficit during summer correlated with substantial increase in δ13C of leaf and phloem sap soluble sugars. Increases in δ13C of ecosystem respired CO2 were linearly related to increases in phloem sugar δ13C (r2=0.99, P≤0.001) and leaf sugar δ13C (r2=0.981, P≤0.01), indicating that a major proportion of ecosystem respired CO2 was derived from recent assimilates. The slopes of the best-fit lines differed significantly (P≤0.05), however, and were about 0.86 (SE=0.04) for phloem sugars and about 1.63 (SE=0.16) for leaf sugars. Hence, changes in isotopic signature in phloem sugars were transferred to ecosystem respiration in the beech forest, while leaf sugars, with relatively small seasonal changes in δ13C, must have a slower turnover rate or a significant storage component. No significant variation in δ13C was observed in bulk dry matter of various plant and ecosystem components (including leaves, bark, wood, litter and soil organics). The apparent coupling between the δ13C of soluble sugars and ecosystem respiration was associated with large apparent isotopic disequilibria. Values of δ13CR were consistently more depleted by about 4‰ relative to phloem sugars, and by about 2‰ compared to leaf sugars. Since no combination of the measured pools could produce the observed δ13CR signal over the entire season, a significant isotopic discrimination against 13C might be associated with short-term ecosystem respiration. However, these differences might also be explained by substantial contributions of other not measured carbon pools (e.g., lipids) to ecosystem respiration or contributions linked to differences in footprint area between Keeling plots and carbohydrate sampling. Linking the seasonal and inter-annual variations in carbon isotope composition of carbohydrates and respiratory CO2 should be applicable in carbon cycle models and help the understanding of inter-annual variation in biospheric sink strength.

Seasonal and inter-annual dynamics of growth, non-structural carbohydrates and C stable isotopes in a Mediterranean beech forest.

Seasonal and inter-annual dynamics of growth, non-structural carbohydrates (NSC) and carbon isotope composition (δ(13)C) of NSC were studied in a beech forest of Central Italy over a 2-year period characterized by different environmental conditions. The net C assimilated by forest trees was mainly used to sustain growth early in the season and to accumulate storage carbohydrates in trunk and root wood in the later part of the season, before leaf shedding. Growth and NSC concentration dynamics were only slightly affected by the reduced soil water content (SWC) during the drier year. Conversely, the carbon isotope analysis on NSC revealed seasonal and inter-annual variations of photosynthetic and post-carboxylation fractionation processes, with a significant increase in δ(13)C of wood and leaf soluble sugars in the drier summer year than in the wetter one. The highly significant correlation between δ(13)C of leaf soluble sugars and SWC suggests a decrease of the canopy C isotope discrimination and, hence, an increased water-use efficiency with decreasing soil water availability. This may be a relevant trait for maintaining an acceptable plant water status and a relatively high C sink capacity during dry seasonal periods. Our results suggest a short- to medium-term homeostatic response of the Collelongo beech stand to variations in water availability and solar radiation, indicating that this Mediterranean forest was able to adjust carbon-water balance in order to prevent C depletion and to sustain plant growth and reserve accumulation during relatively dry seasons.

Investigating the European beech (Fagus sylvatica L.) leaf characteristics along the vertical canopy profile: leaf structure, photosynthetic capacity, light energy dissipation and photoprotection mechanisms

Forest functionality and productivity are directly related to canopy light interception and can be affected by potential damage from high irradiance. However, the mechanisms by which leaves adapt to the variable light environments along the multilayer canopy profile are still poorly known. We explored the leaf morphophysiological and metabolic responses to the natural light gradient in a pure European beech (Fagus sylvatica L.) forest at three different canopy heights (top, middle and bottom). Structural adjustment through light-dependent modifications in leaf mass per area was the reason for most of the variations in photosynthetic capacity. The different leaf morphology along the canopy influenced nitrogen (N) partitioning, water- and photosynthetic N-use efficiency, chlorophyll (Chl) fluorescence and quali-quantitative contents of photosynthetic pigments. The Chl a to Chl b ratio and the pool of xanthophyll-cycle pigments (VAZ) increased at the highest irradiance, as well as lutein and β-carotene. The total pool of ascorbate and phenols was higher in leaves of the top and middle canopy layers when compared with the bottom layer, where the ascorbate peroxidase was relatively more activated. The non-photochemical quenching was strongly and positively related to the VAZ/(Chl a + b) ratio, while Chl a/Chl b was related to the photochemical efficiency of photosystem II. Along the multilayer canopy profile, the high energy dissipation capacity of leaves was correlated to an elevated redox potential of antioxidants. The middle layer gave the most relevant contribution to leaf area index and carboxylation capacity of the canopy. In conclusion, a complex interplay among structural, physiological and biochemical traits drives the dynamic leaf acclimation to the natural gradients of variable light environments along the tree canopy profile. The relevant differences observed in leaf traits within the canopy positions of the beech forest should be considered for improving estimation of carbon fluxes in multilayer canopy models of temperate forests.

Comparing integrated stable isotope and eddy covariance estimates of water-use efficiency on a Mediterranean successional sequence

Water-use efficiency (WUE), thought to be a relevant trait for productivity and adaptation to water-limited environments, was estimated for three different ecosystems on the Mediterranean island of Pianosa: Mediterranean macchia (SMM), transition (STR) and abandoned agricultural (SAA) ecosystems, representing a successional series. Three independent approaches were used to study WUE: eddy covariance measurements, C isotope composition of ecosystem respired CO2, and C isotope discrimination (Δ) of leaf material (dry matter and soluble sugars). Seasonal variations in C–water relations and energy fluxes, compared in SMM and in SAA, were primarily dependent on the specific composition of each plant community. WUE of gross primary productivity was higher in SMM than in SAA at the beginning of the dry season. Both structural and fast-turnover leaf material were, on average, more enriched in 13C in SMM than SAA, indicating relatively higher stomatal control and WUE for the long-lived macchia species. This pattern corresponded to 13C-enriched respired CO2 in SMM compared to the other ecosystems. Conversely, most of the annual herbaceous SAA species (terophytes) showed a drought-escaping strategy, with relatively high stomatal conductance and low WUE. An ecosystem-integrated Δ value was weighted for each ecosystem on the abundance of different life forms, classified according to Raunkiar’s system. Agreement was found between ecosystem WUE calculated using eddy covariance and those estimated using integrated Δ approaches. Comparing the isotopic methods, Δ of leaf soluble sugars provided the most reliable proxy for short-term changes in photosynthetic discrimination and associated shifts in integrated canopy-level WUE along the successional series.

Trichoderma harzianum T6776 modulates a complex metabolic network to stimulate tomato cv. Micro-Tom growth

Background and aimsTrichoderma harzianum 6776 is a novel and beneficial tomato fungal isolate. To investigate the mechanisms underlying the T. harzianum 6776-tomato interaction, several physiological and biochemical responses were explored on dwarf tomato plants, cv. Micro-Tom.MethodsGrowth of treated and untreated plants was evaluated by measuring the height and biomass production of plants. The leaf pigment content and sugar partitioning in plant organs were evaluated by biochemical analysis. The photosynthetic parameters were measured by a miniaturized PAM fluorometer and a portable gas-exchange system. The hormonal analysis in root and xylem sap was performed by gas cromatography- mass spectrometry (GC-MS).ResultsT. harzianum 6776 positively affected plant growth, increasing the leaf pigment content and improving the photosynthetic activity at both stomatal and non-stomatal levels. Differences in pigment composition and photosynthetic performance were reflected in the carbohydrate content and their partitioning. In the absence of a pathogen, root and xylem vessel stress and growth-related hormone balance were affected by the interaction with T. harzianum 6776, with an increase in jasmonic and indoleacetic acids and a decrease in salicylic acid content.ConclusionsThis study shows the complex connection between increased hormone accumulation and transport, altered sugar partitioning and enhanced photosynthetic efficiency induced by T. harzianum 6776, and how growth promotion is the result of the combination of these drastic changes in Micro-Tom plants.

Effect of carbon/nitrogen ratio on carbohydrate metabolism and light energy dissipation mechanisms in Arabidopsis thaliana.

Carbon (C) and nitrogen (N) nutrient sources are essential elements for metabolism, and their availability must be tightly coordinated for the optimal growth and development in plants. Plants are able to sense and respond to different C/N conditions via specific partitioning of C and N sources and the regulation of a complex cellular metabolic activity. We studied how the interaction between C and N signaling could affect carbohydrate metabolism, soluble sugar levels, photochemical efficiency of photosystem II (PSII) and the ability to drive the excess energy in Arabidopsis seedlings under moderated and disrupted C/N-nutrient conditions. Invertase and sucrose synthase activities were markedly affected by C/N-nutrient status depending on the phosphorylation status, suggesting that these enzymes may necessarily be modulated by their direct phosphorylation or phosphorylation of proteins that form complex with them in response to C/N stress. In addition, the enzymatic activity of these enzymes was also correlated with the amount of sugars, which not only act as substrate but also as signaling compounds. Analysis of chlorophyll fluorescence in plants under disrupted C/N condition suggested a reduction of electron transport rate at PSII level associated with a higher capacity for non-radiative energy dissipation in comparison with plants under moderated C/N condition. In conclusion, the tight coordination between C and N not only affects the carbohydrates metabolism and their concentration within plant tissues, but also the partitioning of the excitation energy at PSII level between radiative (electron transport) and non-radiative (heat) dissipation pathways.

Combining stable isotope and carbohydrate analyses in phloem sap and fine roots to study seasonal changes of source-sink relationships in a Mediterranean beech forest.

Carbon isotope composition (δ(13)C) and carbohydrate content of phloem sap and fine roots were measured in a Mediterranean beech (Fagus sylvatica L.) forest throughout the growing season to study seasonal changes of source-sink relationships. Seasonal variations of δ(13)C and content of phloem sap sugars, collected during the daylight period, reflected the changes in soil and plant water status. The correlation between δ(13)C and content of phloem sap sugars, collected from plants belonging to different social classes, was significantly positive only during the driest month of July. In this month, δ(13)C of phloem sap sugars was inversely related to the increment of trunk radial growth and positively related to δ(13)C of fine roots. We conclude that the relationship between δ(13)C and the amount of phloem sap sugars is affected by a combination of causes, such as sink strength, tree social class, changes in phloem anatomy and transport capacity, and phloem loading of sugars to restore sieve tube turgor following the reduced plant water potential under drought conditions. However, δ(13)C and sugar composition of fine roots suggested that phloem transport of leaf sucrose to this belowground component was not impaired by mild drought and that sucrose was in a large part allocated towards fine roots in July, depending on tree social class. Hence, fine roots could represent a functional carbon sink during the dry seasonal periods, when transport and use of assimilates in other sink tissues are reduced. These results indicate a strict link between above- and belowground processes and highlight a rapid response of this Mediterranean forest to changes in environmental drivers to regulate source-sink relationships and carbon sink capacity.

Carbon and nitrogen allocation strategy in Posidonia oceanica is altered by seawater acidification

Rising atmospheric CO2 causes ocean acidification that represents one of the major ecological threats for marine biota. We tested the hypothesis that long-term exposure to increased CO2 level and acidification in a natural CO2 vent system alters carbon (C) and nitrogen (N) metabolism in Posidonia oceanica L. (Delile), affecting its resilience, or capability to restore the physiological homeostasis, and the nutritional quality of organic matter available for grazers. Seawater acidification decreased the C to N ratio in P. oceanica tissues and increased grazing rate, shoot density, leaf proteins and asparagine accumulation in rhizomes, while the maximum photochemical efficiency of photosystem II was unaffected. The 13C-dilution in both structural and non-structural C metabolites in the acidified site indicated quali-quantitative changes of C source and/or increased isotopic fractionation during C uptake and carboxylation associated with the higher CO2 level. The decreased C:N ratio in the acidified site suggests an increased N availability, leading to a greater storage of 15N-enriched compounds in rhizomes. The amount of the more dynamic C storage form, sucrose, decreased in rhizomes of the acidified site in response to the enhanced energy demand due to higher shoot recruitment and N compound synthesis, without affecting starch reserves. The ability to modulate the balance between stable and dynamic C reserves could represent a key ecophysiological mechanism for P. oceanica resilience under environmental perturbation. Finally, alteration in C and N dynamics promoted a positive contribution of this seagrass to the local food web.

The role of Euglena gracilis paramylon in modulating xylem hormone levels, photosynthesis and water-use efficiency in Solanum lycopersicum L.

β-1,3-glucans such as paramylon act as elicitors in plants, modifying the hormonal levels and the physiological responses. Plant hormones affect all phases of the plant life cycle and their responses to environmental stresses, both biotic and abiotic. The aim of this study was to investigate the effects of a root treatment with Euglena gracilis paramylon on xylem hormonal levels, photosynthetic performance and dehydration stress in tomato (Solanum lycopersicum). Paramylon granules were processed to obtain the linear fibrous structures capable to interact with tomato cell membrane. Modulation of hormone levels (abscisic acid, jasmonic acid and salicylic acid) and related physiological responses such as CO2 assimilation rate, stomatal and mesophyll conductance, intercellular CO2 concentration, transpiration rate, water-use efficiency, quantum yield of photosystem II and leaf water potential were investigated. The results indicate a clear dose-dependent effect of paramylon on the hormonal content of xylem sap, photosynthetic performance and dehydration tolerance. Paramylon has the capability to enhance plant defense capacity against abiotic stress, such as drought, by modulating the conductance to CO2 diffusion from air to the carboxylation sites and improving the water-use efficiency.

Late summer photosynthesis and storage carbohydrates in walnut (Juglans regia L.): Feed-back and feed-forward effects

The effect of late summer - autumn limitation of phloem export on growth, photosynthesis and storage carbohydrate accumulation, was evaluated in walnut (Juglans regia L.). This was done by girdling current years shoots, with either all or with only a third of the leaves left in place. Nineteen days after girdling, photosynthesis was greatly reduced and after 46 days, it was about 70% lower in both girdling treatments compared to the control (ungirdled shoots). This reduction is consistent with a feed-back effect of an increased carbohydrate content of the leaves. At the end of the experiment (46 days after girdling), the radial growth of girdled shoots was increased at their base but not at their apical part compared to the control. Girdling increased the accumulation of sucrose in the bark at the base of the shoot and of starch in the bark and in the wood of the shoot apical part. The activity of ADP-glucose pyrophosphorylase in wood increased in the apical part of girdled shoots. The results suggest that a high availability of carbohydrates elicits a feed-forward action on the shoot sink size and activity (radial growth and storage carbohydrate accumulation). Further, for the first time in tree wood we found an increased total activity of AGP induced by an increased assimilate availability. Moreover, the results indicated that, in late summer - autumn, CO2 uptake by leaves of the deciduous tree walnut is strongly dependent on export of photosynthates from the crown. Therefore, carbon uptake in this period depends largely on the availability of effective storage sinks where newly produced assimilates can be accumulated.