Plant and microbial science and technology as cornerstones to Bioregenerative Life Support Systems in space.

Long-term human space exploration missions require environmental control and closed Life Support Systems (LSS) capable of producing and recycling resources, thus fulfilling all the essential metabolic needs for human survival in harsh space environments, both during travel and on orbital/planetary stations. This will become increasingly necessary as missions reach farther away from Earth, thereby limiting the technical and economic feasibility of resupplying resources from Earth. Further incorporation of biological elements into state-of-the-art (mostly abiotic) LSS, leading to bioregenerative LSS (BLSS), is needed for additional resource recovery, food production, and waste treatment solutions, and to enable more self-sustainable missions to the Moon and Mars. There is a whole suite of functions crucial to sustain human presence in Low Earth Orbit (LEO) and successful settlement on Moon or Mars such as environmental control, air regeneration, waste management, water supply, food production, cabin/habitat pressurization, radiation protection, energy supply, and means for transportation, communication, and recreation. In this paper, we focus on air, water and food production, and waste management, and address some aspects of radiation protection and recreation. We briefly discuss existing knowledge, highlight open gaps, and propose possible future experiments in the short-, medium-, and long-term to achieve the targets of crewed space exploration also leading to possible benefits on Earth.

Perspectives for plant biology in space and analogue environments.

Advancements in plant space biology are required for the realization of human space exploration missions, where the re-supply of resources from Earth is not feasible. Until a few decades ago, space life science was focused on the impact of the space environment on the human body. More recently, the interest in plant space biology has increased because plants are key organisms in Bioregenerative Life Support Systems (BLSS) for the regeneration of resources and fresh food production. Moreover, plants play an important role in psychological support for astronauts. The definition of cultivation requirements for the design, realization, and successful operation of BLSS must consider the effects of space factors on plants. Altered gravitational fields and radiation exposure are the main space factors inducing changes in gene expression, cell proliferation and differentiation, signalling and physiological processes with possible consequences on tissue organization and organogenesis, thus on the whole plant functioning. Interestingly, the changes at the cellular and molecular levels do not always result in organismic or developmental changes. This apparent paradox is a current research challenge. In this paper, the main findings of gravity- and radiation-related research on higher plants are summarized, highlighting the knowledge gaps that are still necessary to fill. Existing experimental facilities to simulate the effect of space factors, as well as requirements for future facilities for possible experiments to achieve fundamental biology goals are considered. Finally, the need for making synergies among disciplines and for establishing global standard operating procedures for analyses and data collection in space experiments is highlighted.

Defining growth requirements of microgreens in space cultivation via biomass production, morpho-anatomical and nutritional traits analysis.

During long-term manned missions to the Moon or Mars, the integration of astronauts' diet with fresh food rich in functional compounds, like microgreens, could strengthen their physiological defenses against the oxidative stress induced by the exposure to space factors. Therefore, the development of targeted cultivation practices for microgreens in space is mandatory, since the cultivation in small, closed facilities may alter plant anatomy, physiology, and resource utilization with species-specific responses. Here, the combined effect of two vapor pressure deficit levels (VPD: 0.14 and 1.71 kPa) and two light intensities (150 and 300 µmol photons m-2 s-1 PPFD) on two species for microgreen production (Brassica oleracea var. capitata f. sabauda 'Vertus' and Raphanus raphanistrum subsp. sativus 'Saxa'), was tested on biomass production per square meter, morpho-anatomical development, nutritional and nutraceutical properties. Microgreens were grown in fully controlled conditions under air temperature of 18/24°C, on coconut fiber mats, RGB light spectrum and 12 h photoperiod, till they reached the stage of first true leaves. At this stage microgreens were samples, for growth and morpho-anatomical analyses, and to investigate the biochemical composition in terms of ascorbic acid, phenols, anthocyanin, carotenoids, carbohydrates, as well as of anti-nutritional compounds, such as nitrate, sulfate, and phosphate. Major differences in growth were mostly driven by the species with 'Saxa' always presenting the highest fresh and dry weight as well as the highest elongation; however light intensity and VPDs influenced the anatomical development of microgreens, and the accumulation of ascorbic acid, carbohydrates, nitrate, and phosphate. Both 'Saxa' and 'Vertus' at low VPD (LV) and 150 PPFD increased the tissue thickness and synthetized high ß-carotene and photosynthetic pigments. Moreover, 'Vertus' LV 150, produced the highest content of ascorbate, fundamental for nutritional properties in space environment. The differences among the treatments and their interaction suggested a relevant difference in resource use efficiency. In the light of the above, microgreens can be considered suitable for cultivation in limited-volume growth modules directly onboard, provided that all the environmental factors are combined and modulated according to the species requirements to enhance their growth and biomass production, and to achieve specific nutritional traits.

Radiation environment in exploration-class space missions and plants' responses relevant for cultivation in Bioregenerative Life Support Systems.

For deep space exploration, radiation effects on astronauts, and on items fundamental for life support systems, must be kept under a pre-agreed threshold to avoid detrimental outcomes. Therefore, it is fundamental to achieve a deep knowledge on the radiation spatial and temporal variability in the different mission scenarios as well as on the responses of different organisms to space-relevant radiation. In this paper, we first consider the radiation issue for space exploration from a physics point of view by giving an overview of the topics related to the spatial and temporal variability of space radiation, as well as on measurement and simulation of irradiation, then we focus on biological issues converging the attention on plants as one of the fundamental components of Bioregenerative Life Support Systems (BLSS). In fact, plants in BLSS act as regenerators of resources (i.e. oxygen production, carbon dioxide removal, water and wastes recycling) and producers of fresh food. In particular, we summarize some basic statements on plant radio-resistance deriving from recent literature and concentrate on endpoints critical for the development of Space agriculture. We finally indicate some perspective, suggesting the direction future research should follow to standardize methods and protocols for irradiation experiments moving towards studies to validate with space-relevant radiation the current knowledge. Indeed, the latter derives instead from experiments conducted with different radiation types and doses and often with not space-oriented scopes.

Integration of high-throughput phenotyping with anatomical traits of leaves to help understanding lettuce acclimation to a changing environment.

MAIN CONCLUSION: The combination of image-based phenotyping with in-depth anatomical analysis allows for a thorough investigation of plant physiological plasticity in acclimation, which is driven by environmental conditions and mediated by anatomical traits. Understanding the ability of plants to respond to fluctuations in environmental conditions is critical to addressing climate change and unlocking the agricultural potential of crops both indoor and in the field. Recent studies have revealed that the degree of eco-physiological acclimation depends on leaf anatomical traits, which show stress-induced alterations during organogenesis. Indeed, it is still a matter of debate whether plant anatomy is the bottleneck for optimal plant physiology or vice versa. Here, we cultivated 'Salanova' lettuces in a phenotyping chamber under two different vapor pressure deficits (VPDs; low, high) and watering levels (well-watered, low-watered); then, plants underwent short-term changes in VPD. We aimed to combine high-throughput phenotyping with leaf anatomical analysis to evaluate their capability in detecting the early stress signals in lettuces and to highlight the different degrees of plants' eco-physiological acclimation to the change in VPD, as influenced by anatomical traits. The results demonstrate that well-watered plants under low VPD developed a morpho-anatomical structure in terms of mesophyll organization, stomatal and vein density, which more efficiently guided the acclimation to sudden changes in environmental conditions and which was not detected by image-based phenotyping alone. Therefore, we emphasized the need to complement high-throughput phenotyping with anatomical trait analysis to unveil crop acclimation mechanisms and predict possible physiological behaviors after sudden environmental fluctuations due to climate changes.

Vapour Pressure Deficit (VPD) Drives the Balance of Hydraulic-Related Anatomical Traits in Lettuce Leaves.

The coordination of leaf hydraulic-related traits with leaf size is influenced by environmental conditions and especially by VPD. Water and gas flows are guided by leaf anatomical and physiological traits, whose plasticity is crucial for plants to face environmental changes. Only a few studies have analysed how variations in VPD levels influence stomatal and vein development and their correlation with leaf size, reporting contrasting results. Thus, we applied microscopy techniques to evaluate the effect of low and high VPDs on the development of stomata and veins, also analysing leaf functional traits. We hypothesized that leaves under high VPD with a modified balance between veins and stomata face higher transpiration. We also explored the variability of stomata and vein density across the leaf lamina. From the results, it was evident that under both VPDs, plants maintained a coordinated development of stomata and veins, with a higher density at low VPD. Moreover, more stomata but fewer veins developed in the parts of the lettuce head exposed to light, suggesting that their differentiation during leaf expansion is strictly dependent on the microclimatic conditions. Knowing the plasticity of hydraulic-related morpho-functional traits and its intra-leaf variability is timely for their impact on water and gas fluxes, thus helping to evaluate the impact of environmental-driven anatomical variations on productivity of natural ecosystems and crops, in a climate change scenario.

Morpho-Anatomical, Physiological, and Mineral Composition Responses Induced by a Vegetal-Based Biostimulant at Three Rates of Foliar Application in Greenhouse Lettuce.

A promising strategy for sustainably increasing the quality and yield of horticultural products is the use of natural plant biostimulants. In this work, through a greenhouse experiment, we evaluated the effect of a legume-derived biostimulant at three dose treatments (0.0 control, 2.5 mL L-1, and 5.0 mL L-1) on the yield performance, nutrients traits, leaf anatomical traits, gas exchanges, and carbon photosynthetic assimilation of greenhouse lettuce. The lettuce plants were foliar sprayed every 7 days for 5 weeks. The application of plant biostimulant, at both lower and higher dosages, increased the nutrient use efficiency, root dry weight, and leaf area. However, it is noteworthy that the 5.0 mL L-1 dose enhanced photosynthetic activity in the early phase of growth (15 DAT), thus supplying carbon skeletons useful for increasing the number of leaves and their efficiency (higher SPAD), and for boosting nutrient uptake (P, S, and K) and transport to leaves, while the 2.5 mL L-1 dose exerted specific effects on roots, increasing their dimension and enabling them to better use nitrate and Ca. A higher dose of biostimulant application might find its way in shorter growing cycle, thus presenting new horizons for new lines of research in baby leaves production.

Light Quality Modulates Photosynthesis and Antioxidant Properties of B. vulgaris L. Plants from Seeds Irradiated with High-Energy Heavy Ions: Implications for Cultivation in Space.

Beta vulgaris L. is a crop selected for cultivation in Space for its nutritional properties. However, exposure to ionizing radiation (IR) can alter plant photosynthetic performance and phytochemical production in the extraterrestrial environment. This study investigated if plant growth under different light quality regimes (FL-white fluorescent; RGB-red-green-blue; RB-red-blue) modifies the photosynthetic behavior and bioactive compound synthesis of plants sprouted by dry seeds irradiated with carbon or titanium high-energy ions. The study evidenced that: (i) the plant response depends on the type of heavyion; (ii) control and C-ion-irradiated plants were similar for photosynthetic pigment content and PSII photochemical efficiency, regardless of the LQ regime; (iii) under FL, net photosynthesis (AN) and water use efficiency (iWUE) declined in C- and Ti-ion plants compared to control, while the growth of irradiated plants under RGB and RB regimes offset these differences; (iv) the interaction Ti-ion× RB improved iWUE, and stimulated the production of pigments, carbohydrates, and antioxidants. The overall results highlighted that the cultivation of irradiated plants under specific LQ regimes effectively regulates photosynthesis and bioactive compound amounts in leaf edible tissues. In particular, the interaction Ti-ion × RB improved iWUE and increased pigments, carbohydrates, and antioxidant content.

Field evidence for litter and self-DNA inhibitory effects on Alnus glutinosa roots.

Litter decomposition releases nutrients beneficial to plants but also induces phytotoxicity. Phytotoxicity can result from either labile allelopathic compounds or species specific and caused by conspecific DNA. Aquatic plants in flowing water generally do not suffer phytotoxicity because litter is regularly removed. In stagnant water or in litter packs an impact on root functionality can occur. So far, studies on water plant roots have been carried out in laboratory and never in field conditions. The effect of conspecific vs heterospecific litter and purified DNA were assessed on aquatic roots of the riparian woody species Alnus glutinosa L. using a novel method, using closed and open plastic tubes fixed to single roots in the field with closed tubes analogous to stagnant water. Four fresh and four decomposed litter types were used and analysed on extractable C, cellulose, lignin, N content and using 13 C-CPMAS NMR spectroscopy. Inhibitory effects were observed with fresh litter in closed systems, with a positive correlation with extractable C and negative with lignin and lignin : N ratio. Alnus self-DNA, but not heterologous one, caused acute toxic effects in the closed system. Our results demonstrate the first field-based evidence for self-DNA inhibition as causal factor of negative feedback between plants and substrate.

How Leaf Vein and Stomata Traits Are Related with Photosynthetic Efficiency in Falanghina Grapevine in Different Pedoclimatic Conditions.

The increase in severe drought events due to climate change in the areas traditionally suitable for viticulture is enhancing the need to understand how grapevines regulate their photosynthetic metabolism in order to forecast specific cultivar adaptive responses to the changing environment. This study aims at evaluating the association between leaf anatomical traits and eco-physiological adjustments of the 'Falanghina' grapevine under different microclimatic conditions at four sites in southern Italy. Sites were characterized by different pedoclimatic conditions but, as much as possible, were similar for plant material and cultivation management. Microscopy analyses on leaves were performed to quantify stomata and vein traits, while eco-physiological analyses were conducted on vines to assess plant physiological adaptation capability. At the two sites with relatively low moisture, photosynthetic rate, stomatal conductance, photosystem electron transfer rate, and quantum yield of PSII, linear electron transport was lower compared to the other two sites. Stomata size was higher at the site characterized by the highest precipitation. However, stomatal density and most vein traits tended to be relatively stable among sites. The number of free vein endings per unit leaf area was lower in the two vineyards with low precipitation. We suggest that site-specific stomata and vein traits modulation in Falanghina grapevine are an acclimation strategy that may influence photosynthetic performance. Overall in-depth knowledge of the structure/function relations in Falanghina vines might be useful to evaluate the plasticity of this cultivar towards site-specific management of vineyards in the direction of precision viticulture.

Comparative Analysis of the Effect of Carbon- and Titanium-Ions Irradiation on Morpho-Anatomical and Biochemical Traits of Dolichos melanophthalmus DC. Seedlings Aimed to Space Exploration.

The realization of manned missions for space exploration requires the development of Bioregenerative Life Support Systems (BLSSs) to make human colonies self-sufficient in terms of resources. Indeed, in these systems, plants contribute to resource regeneration and food production. However, the cultivation of plants in space is influenced by ionizing radiation which can have positive, null, or negative effects on plant growth depending on intrinsic and environmental/cultivation factors. The aim of this study was to analyze the effect of high-LET (Linear Energy Transfer) ionizing radiation on seed germination and seedling development in eye bean. Dry seeds of Dolichos melanophthalmus DC. (eye bean) were irradiated with two doses (1 and 10 Gy) of C- and Ti-ions. Seedlings from irradiated seeds were compared with non-irradiated controls in terms of morpho-anatomical and biochemical traits. Results showed that the responses of eye bean plants to radiation are dose-specific and dependent on the type of ion. The information obtained from this study will be useful for evaluating the radio-resistance of eye bean seedlings, for their possible cultivation and utilization as food supplement in space environments.

Light Spectral Composition Influences Structural and Eco-Physiological Traits of Solanum lycopersicum L. cv. 'Microtom' in Response to High-LET Ionizing Radiation.

This study evaluated if specific light quality (LQ) regimes (white fluorescent, FL; full-spectrum, FS; red-blue, RB) during plant growth modified morphological and photosynthetic traits of Solanum lycopersicum L. 'Microtom' plants irradiated at the dry seed stage with 25 Gy 48Ca ions (IR). The irradiation reduced plant size while it increased leaf dry matter content (LDMC) and relative water content (RWC) compared to the control. FS and RB light regimes determined a decrease of plant height and a rise of RWC compared to FL plants. The irradiation under FS and RB regimes favoured the development of dwarf plants and improved the leaf water status. Under the FL regime, irradiated plants showed reduced photosynthesis and stomatal conductance. The opposite behavior was observed in RB irradiated plants in which gas exchanges were significantly stimulated. RB regime enhanced Rubisco expression in irradiated plants also inducing anatomical and functional adjustments (i.e., increase of leaf thickness and incidence of intercellular spaces). Finally, 48Ca ions did not prevent fruit ripening and the achievement of the 'seed-to seed' cycle, irrespective of the LQ regime. Overall, the present study evidenced that RB light regime was the most effective in optimising growth and photosynthetic efficiency of 'Microtom' irradiated plants. These outcomes may help to develop proper cultivation protocols for the growth of dwarf tomato in Controlled Ecological Life Support Systems (CELSS).

Isosmotic Macrocation Variation Modulates Mineral Efficiency, Morpho-Physiological Traits, and Functional Properties in Hydroponically Grown Lettuce Varieties (Lactuca sativa L.).

The management of mineral elements in agriculture is important for their nutritional role for plants and dietary value for humans, sparking interest in strategies that can increase mineral use efficiency and accumulation in plant food. In this work, we evaluated the effects of the isosmotic variations of the concentration on three macrocations (K, Ca, and Mg) in lettuce (Lactuca sativa L.). Our aim was to improve the nutritional components of this valuable dietary source of minerals. Using a full factorial design, we analyzed mineral utilization efficiency (UtE), leaf morphology, gas exchange parameters, phenolic profiles (through ultra-high performance liquid chromatography coupled to a quadrupole-time-of-flight (UHPLC-QTOF) mass spectrometry), and enzymatic activities in two phytochemically diverse butterhead lettuce varieties (red or green). Plants were fed in hydroponics with three nutrient solutions (NSs) with different ratios of K, Ca, and Mg. The variation of these minerals in the edible product was associated with alterations of the morphology and physiology of the leaves, and of the quality and functional properties of lettuce, with a trade-off between total accumulation and mineral UtE. Moreover, in non-limiting conditions of nutrient availability, significant mineral interactions were also present. The flexibility of the plant response to the different ratios of macrocations, and the observed large intraspecific variation, were adequate to provide mineral-specific phytochemical profiles to the edible product. Specifically, the full-red lettuce provided more interesting results in regard to the compositional and functional attributes of the leaves.

Light and Low Relative Humidity Increase Antioxidants Content in Mung Bean (Vigna radiata L.) Sprouts.

In the last decades, there has been a growing interest in the production of sprouts, since they are a highly nutritious food, particularly suitable for indoor farming in urban areas. Achieving sprout production in indoor systems requires an understanding of possible alterations induced by the microclimate. The aim of this study was to analyze the combined effect of presence/absence of light and high/low air relative humidity (RH) on mung bean sprouts. Morpho-anatomical development and functional anatomical traits in hypocotyl were quantified. The content of antioxidants, soluble sugars, and starch were measured for nutritional and functional purposes. Different RH regimes mainly induced morpho-anatomical modifications, while the presence/absence of light changed the content of antioxidant compounds. Increments in stele diameter at high RH suggest a higher water uptake and conductivity, compared to the low RH treatment; low RH and light induced anatomical traits improving plant water transport (reduced number of cortical layers) and increased the production of antioxidants. The overall results suggested that RH and light, already at the early stages of development, affect the plant's nutritional value. Therefore, the combination of light and low RH allows the production of antioxidant-rich mung bean sprouts to be used as a food supplement.

Understanding the Morpho-Anatomical, Physiological, and Functional Response of Sweet Basil to Isosmotic Nitrate to Chloride Ratios.

Sweet basil (Ocimum basilicum L.) is a leafy green with a short-production cycle that is emerging as a model species among aromatic plants. Modulating the mineral composition of the nutrient solution has proved to be a valuable tool to uncover the mechanisms and responses that higher plants adopt in relation to the availability of mineral nutrients. The aim of this work was to examine the effects on basil of four isosmotic nutrient solutions with different nitrate to chloride ratios. These two anions share uptake and transport mechanisms in plants and are often considered antagonist. To this goal, we analyzed morpho-anatomical and physiological parameters as well as quality-related traits, such as the antioxidant capacity, the leaf color, the mineral composition, and the aromatic profile in relation to the nutrient ratios. Moreover, using a full factorial design, we analyzed leaves in two consecutive harvests. The data indicated a broad, multifaceted plant response to the different nutritional ratios, with almost all the recorded parameters involved. Overall, the effects on basil can be explained by considering an interdependent combination of the nitrate and chloride roles in plant nutrition and physiology. Our work revealed the extent of the modification that can be achieved in basil through the modification of the nutrient solution. It also provided indications for more nutrient efficient growing conditions, because a moderate increase in chloride limits the expected negative impact of a sub-optimal nitrate fertilization.

Crop Management in Controlled Environment Agriculture (CEA) Systems Using Predictive Mathematical Models.

Proximal sensors in controlled environment agriculture (CEA) are used to monitor plant growth, yield, and water consumption with non-destructive technologies. Rapid and continuous monitoring of environmental and crop parameters may be used to develop mathematical models to predict crop response to microclimatic changes. Here, we applied the energy cascade model (MEC) on green- and red-leaf butterhead lettuce (Lactuca sativa L. var. capitata). We tooled up the model to describe the changing leaf functional efficiency during the growing period. We validated the model on an independent dataset with two different vapor pressure deficit (VPD) levels, corresponding to nominal (low VPD) and off-nominal (high VPD) conditions. Under low VPD, the modified model accurately predicted the transpiration rate (RMSE = 0.10 Lm-2), edible biomass (RMSE = 6.87 g m-2), net-photosynthesis (rBIAS = 34%), and stomatal conductance (rBIAS = 39%). Under high VPD, the model overestimated photosynthesis and stomatal conductance (rBIAS = 76-68%). This inconsistency is likely due to the empirical nature of the original model, which was designed for nominal conditions. Here, applications of the modified model are discussed, and possible improvements are suggested based on plant morpho-physiological changes occurring in sub-optimal scenarios.

Biochemical, Physiological and Anatomical Mechanisms of Adaptation of Callistemon citrinus and Viburnum lucidum to NaCl and CaCl2 Salinization.

Callistemon citrinus and Viburnum lucidum are appreciated and widespread ornamental shrubs for their abundant flowering and/or brilliant foliage. The intrinsic tolerance to drought/salinity supports their use in urban areas and in xeriscaping. Despite adaptive responses of these ornamental species to sodium chloride (NaCl) have been extensively explored, little is known on the effects of other salt solution, yet iso-osmotic, on their growth, mineral composition and metabolism. The present research was aimed to assess responses at the biochemical, physiological and anatomical levels to iso-osmotic salt solutions of NaCl and CaCl2 to discriminate the effects of osmotic stress and ion toxicity. The two ornamental species developed different salt-tolerance mechanisms depending on the salinity sources. The growth parameters and biomass production decreased under salinization in both ornamental species, independently of the type of salt, with a detrimental effect of CaCl2 on C. citrinus. The adaptive mechanisms adopted by the two ornamental species to counteract the NaCl salinity were similar, and the decline in growth was mostly related to stomatal limitations of net CO2 assimilation rate, together with the reduction in leaf chlorophyll content (SPAD index). The stronger reduction of C. citrinus growth compared to V. lucidum, was due to an exacerbated reduction in net photosynthetic rate, driven by both stomatal and non stomatal limitations. In similar conditions, V. lucidum exhibited other additional adaptive response, such as modification in leaf functional anatomical traits, mostly related to the reduction in the stomata size allowing plants a better control of stomata opening than in C. citrinus. However, C. citrinus plants displayed an increased ability to retain higher Cl- levels in leaves than in roots under CaCl2 salinity compared to V. lucidum, thus, indicating a further attempt to counteract chloride toxicity through an increased vacuolar compartmentalization and to take advantages of them as chip osmotica.

Wood Growth in Pure and Mixed Quercus ilex L. Forests: Drought Influence Depends on Site Conditions.

Climate response of tree-species growth may be influenced by intra- and inter-specific interactions. The different physiological strategies of stress response and resource use among species may lead to different levels of competition and/or complementarity, likely changing in space and time according to climatic conditions. Investigating the drivers of inter- and intra-specific interactions under a changing climate is important when managing mixed and pure stands, especially in a climate change hot spot such as the Mediterranean basin. Mediterranean tree rings show intra-annual density fluctuations (IADFs): the links among their occurrence, anatomical traits, wood growth and stable isotope ratios can help understanding tree physiological responses to drought. In this study, we compared wood production and tree-ring traits in Quercus ilex L. dominant trees growing in two pure and two mixed stands with Pinus pinea at two sites in Southern Italy, on the basis of the temporal variation of cumulative basal area, intrinsic water use efficiency (WUEi), δ18O and IADF frequency in long tree-ring chronologies. The general aim was to assess whether Q. ilex trees growing in pure or mixed stands have a different wood production through time, depending on climatic conditions and stand structure. The occurrence of dry climatic conditions triggered opposite complementarity interactions for Q. ilex growing with P. pinea trees at the two sites. Competitive reduction was experienced at the T site characterized by higher soil water holding capacity (WHC), lower stand density and less steep slope than the S site; on the opposite, high competition occurred at S site. The observed difference in wood growth was accompanied by a higher WUEi due to a higher photosynthetic rate at the T site, while by a tighter stomatal control in mixed stand of S site. IADF frequency in Q. ilex tree rings was linked to higher WUEi, thus to stressful conditions and could be interpreted as strategy to cope with dry periods, independently from the different wood growth. Considering the forecasted water shortage, inter-specific competition should be reduced in denser stands of Q. ilex mixed with P. pinea. Such findings have important implications for forest management of mixed and pure Q. ilex forests.

The Physiological Mechanisms Behind the Earlywood-To-Latewood Transition: A Process-Based Modeling Approach.

In extratropical ecosystems, the growth of trees is cyclic, producing tree rings composed of large-lumen and thin-walled cells (earlywood) alternating with narrow-lumen and thick-walled cells (latewood). So far, the physiology behind wood formation processes and the associated kinetics has rarely been considered to explain this pattern. We developed a process-based mechanistic model that simulates the development of conifer tracheids, explicitly considering the processes of cell enlargement and the deposition and lignification of cell walls. The model assumes that (1) wall deposition gradually slows down cell enlargement and (2) the deposition of cellulose and lignin is regulated by the availability of soluble sugars. The model reliably reproduces the anatomical traits and kinetics of the tracheids of four conifer species. At the beginning of the growing season, low sugar availability in the cambium results in slow wall deposition that allows for a longer enlargement time; thus, large cells with thin walls (i.e., earlywood) are produced. In late summer and early autumn, high sugar availability produces narrower cells having thick cell walls (i.e., latewood). This modeling framework provides a mechanistic link between plant ecophysiology and wood phenology and significantly contributes to understanding the role of sugar availability during xylogenesis.