Dimosthenis Nikolopoulos

The Relationship between Anatomy and Photosynthetic Performance of Heterobaric Leaves

Heterobaric leaves show heterogeneous pigmentation due to the occurrence of a network of transparent areas that are created from the bundle sheaths extensions (BSEs). Image analysis showed that the percentage of photosynthetically active leaf area (Ap) of the heterobaric leaves of 31 plant species was species dependent, ranging from 91% in Malva sylvestris to only 48% inGynerium sp. Although a significant portion of the leaf surface does not correspond to photosynthetic tissue, the photosynthetic capacity of these leaves, expressed per unit of projected area (Pmax), was not considerably affected by the size of their transparent leaf area (At). This means that the photosynthetic capacity expressed per Ap(P*max) should increase with At. Moreover, the expression of P*max could be allowing the interpretation of the photosynthetic performance in relation to some critical anatomical traits. The P*max, irrespective of plant species, correlated with the specific leaf transparent volume (λt), as well as with the transparent leaf area complexity factor (CFAt), parameters indicating the volume per unit leaf area and length/density of the transparent tissues, respectively. Moreover, both parameters increased exponentially with leaf thickness, suggesting an essential functional role of BSEs mainly in thick leaves. The results of the present study suggest that although the Ap of an heterobaric leaf is reduced, the photosynthetic performance of each areole is increased, possibly due to the light transferring capacity of BSEs. This mechanism may allow a significant increase in leaf thickness and a consequent increase of the photosynthetic capacity per unit (projected) area, offering adaptive advantages in xerothermic environments.

“Carbon gain vs. water saving, growth vs. defence”: Two dilemmas with soluble phenolics as a joker

Despite that phenolics are considered as a major weapon against herbivores and pathogens, the primal reason for their evolution may have been the imperative necessity for their UV-absorbing and antioxidant properties in order for plants to compensate for the adverse terrestrial conditions. In dry climates the choice concerning the first dilemma (carbon gain vs. water saving) needs the appropriate structural and metabolic modulations, which protect against stresses such as high UV and visible radiation or drought, but reduce photosynthesis and increase oxidative pressure. Thus, when water saving is chosen, priority is given to protection (including phenolic synthesis), instead of carbon gain and hence growth. At the global level, the different choices by the individual species are expressed by an interspecific negative relationship between total phenolics and photosynthesis. On the other hand, the accumulation of phenolics in water saving plants offers additional defensive functions because these multifunctional compounds can also act as pro-oxidant, antifeeding or toxic factors. Therefore phenolics, as biochemical jokers, can give the answer to both dilemmas: water saving involves high concentrations of phenolics which also offer high level of defence.

Alarm Photosynthesis: Calcium Oxalate Crystals as an Internal CO2 Source in Plants

A new photosynthetic path named “alarm photosynthesis” uses mesophyll calcium oxalate crystals as the CO2 source when stomata are closed, providing adaptive advantages under drought conditions. Calcium oxalate crystals are widespread among animals and plants. In land plants, crystals often reach high amounts, up to 80% of dry biomass. They are formed within specific cells, and their accumulation constitutes a normal activity rather than a pathological symptom, as occurs in animals. Despite their ubiquity, our knowledge on the formation and the possible role(s) of these crystals remains limited. We show that the mesophyll crystals of pigweed (Amaranthus hybridus) exhibit diurnal volume changes with a gradual decrease during daytime and a total recovery during the night. Moreover, stable carbon isotope composition indicated that crystals are of nonatmospheric origin. Stomatal closure (under drought conditions or exogenous application of abscisic acid) was accompanied by crystal decomposition and by increased activity of oxalate oxidase that converts oxalate into CO2. Similar results were also observed under drought stress in Dianthus chinensis, Pelargonium peltatum, and Portulacaria afra. Moreover, in A. hybridus, despite closed stomata, the leaf metabolic profiles combined with chlorophyll fluorescence measurements indicated active photosynthetic metabolism. In combination, calcium oxalate crystals in leaves can act as a biochemical reservoir that collects nonatmospheric carbon, mainly during the night. During the day, crystal degradation provides subsidiary carbon for photosynthetic assimilation, especially under drought conditions. This new photosynthetic path, with the suggested name “alarm photosynthesis,” seems to provide a number of adaptive advantages, such as water economy, limitation of carbon losses to the atmosphere, and a lower risk of photoinhibition, roles that justify its vast presence in plants.

Modification of water entry (xylem vessels) and water exit (stomata) orchestrates long term drought acclimation of wheat leaves

Immediate leaf functional responses to drought, such as stomatal closure and photosynthetic rate reduction, are already known from short-term studies. We tested the hypothesis that long-term acclimation of leaves to drought includes hydraulic and stomatal anatomical changes and that gas exchange and nitrogen allocation patterns are inevitably adjusted to the new structural status. 26 structural and functional traits in one sensitive cultivar (Simeto) and two drought resistant landraces (Ntopia Heraklion 184, Kontopouli 17) of field- grown wheat (Triticum turgidum L. var. durum) were examined under four water shortage levels. Drought acclimation responses were more intense in Simeto than in the two landraces. In accordance to the working hypothesis, drought-acclimated leaves showed lower hydraulic conductance due to narrower vessels and higher stomatal and vein densities than the control leaves, resulting in a safe mode of water transfer and consumption which is essential for the survival in water limiting conditions.Irrespectively of genotype and water regime, significant correlations among structural (hydraulic characteristics, stomatal and vein patterns) and functional (gas exchange, nitrogen content) parameters were found, indicating the functional adjustment to the new structural status. The Principal Component Analysis showed that these structure-function interactions reflected the trade-off between growth and protection against water losses (Axis 1), as well as the competition between different sinks (carbon gain vs structural reinforcement and reproductive effort) in N allocation (Axis 2). Drought acclimation in wheat leaf is integrally processed by the coordination of structural and functional parameters in order to compensate for the adverse effects of water shortage. This structure—function network that regulates the transition from normal growth-mode to protection- mode, includes at least two important “nodal points”: xylem conducting efficiency (water entry) and stomatal function (water exit). This transition also includes the redirection of nitrogen resources to different sinks.

Photosynthetic capacity is negatively correlated with the concentration of leaf phenolic compounds across a range of different species

This study reveals a negative relationship between leaf phenolic compounds and photosynthetic Amax among different plant species. This indicates a functional integration among carbon gain and the concentration of leaf phenolic compounds that reflects the trade-off between growth and defence/protection demands.

Cessation of photosynthesis in Lotus japonicus leaves leads to reprogramming of nodule metabolism

Symbiotic nitrogen fixation (SNF) involves global changes in gene expression and metabolite accumulation in both rhizobia and the host plant. In order to study the metabolic changes mediated by leaf–root interaction, photosynthesis was limited in leaves by exposure of plants to darkness, and subsequently gene expression was profiled by real-time reverse transcription–PCR (RT–PCR) and metabolite levels by gas chromatography–mass spectrometry in the nodules of the model legume Lotus japonicus. Photosynthetic carbon deficiency caused by prolonged darkness affected many metabolic processes in L. japonicus nodules. Most of the metabolic genes analysed were down-regulated during the extended dark period. In addition to that, the levels of most metabolites decreased or remained unaltered, although accumulation of amino acids was observed. Reduced glycolysis and carbon fixation resulted in lower organic acid levels, especially of malate, the primary source of carbon for bacteroid metabolism and SNF. The high amino acid concentrations together with a reduction in total protein concentration indicate possible protein degradation in nodules under these conditions. Interestingly, comparisons between amino acid and protein content in various organs indicated systemic changes in response to prolonged darkness between nodulated and non-nodulated plants, rendering the nodule a source organ for both C and N under these conditions.

Leaf anatomy affects the extraction of photosynthetic pigments by DMSO.

Dimethylsulfoxide (DMSO) is a widely used solvent for the extraction of chlorophylls (Chls) from leaves of higher plants. The method is preferred because the time-consuming steps of grinding and centrifuging are not required and the extracts are stable for a long time period. However, the extraction efficiency of this solvent is not comparable among plant species, whereas the particular leaf anatomical characteristics responsible for this unevenness remain unknown. In order to examine the influence of leaf anatomy on the extraction efficiency of DMSO (i.e. the concentration of Chls extracted with DMSO as % of the concentration of Chls extracted with 80% acetone), leaves of 19 plant species with different anatomical characteristics were incubated for 40min in DMSO at 65 degrees C. Under these conditions, heterobaric leaves, which are characterized by the occurrence of bundle sheath extensions in the mesophyll, showed lower extraction efficiency of DMSO compared to homobaric leaves and conifer needles. Microscopical observations of DMSO incubated leaf tissues showed that bundle sheath extensions behave as anatomical barriers which prevent the diffusion of DMSO within heterobaric leaves, even after prolonged incubation with the solvent. The effect was stronger in heterobaric leaves possessing thick bundle sheath extensions. The extraction efficiency of DMSO in these leaves was improved by vacuum infiltration of the samples in the presence of warm (65 degrees C) solvent.

Distribution profile of stomatal conductance and its interrelations to transpiration rate and water dynamics in young maize laminas under sulfate deprivation

Seven-day-old maize (Zea mays) plants were grown hydroponically for 10 days in S-deprived nutrient solution. The distribution profiles according to the position on the stem of the S-deprived laminas’ stomatal conductance, transpiration rate, photosynthetic rate, dry mass, water content, and specific surface area were monitored relative to control among others. Photochemical efficiency of photosystem II remained unaffected by the deprivation, as well as the specific surface area of all but the embryonic laminas after d2. In S-deficient plants, the embryonic (L0) and the uppermost lamina or the one below it presented mostly significant changes. The response ratios (Rr) of the L0 stomatal conductance oscillated; the oscillation started with an increase at d2. The corresponding Rr values of L0 transpiration and photosynthetic rates started oscillating at d4 in the same fashion. At d8, an increasing gradient appeared in water-content Rr values from L1 to the uppermost lamina. At d10, all but the embryonic laminas presented significantly reduced Rr values in water content. Changes in dry mass and surface area of laminas were synchronized. In control, the transpiration rate expressed per DM unit remained constant during the examined period, while under the deprivation it followed a power function of surface area.

Changes in the properties of calcium-carbon inclusions during leaf development and their possible relationship with leaf functional maturation in three inclusion-bearing species

In many plant species, carbon-calcium inclusion (calcium oxalate crystals or cystoliths containing calcium carbonate) formation is a fundamental part of their physiology even necessary for normal growth and development. Despite the long-standing studies on carbon-calcium inclusions, the alterations in their properties during leaf development and their possible association with the maturation of the photosynthetic machinery have not been previously examined. In order to acquire more insights into this subject, we examined three of the most common species bearing abundant inclusions of different types, i.e., Amaranthus hybridus, Vitis vinifera, and Parietaria judaica. Results of our study showed that, irrespective of species and type of inclusion, similar patterns in the alterations of their properties are observed during leaf maturation, except for some differences in cell differentiation and distribution between raphides and druses in Vitis vinifera. As expected, inclusion formation has taken place at very early developmental stages and maximum density was observed in very young leaves. Inclusion properties are changing in a coordinated way with leaf area and these modifications are compatible with the concept that each idioblast or lithocyst “services” a finite number/area of adjacent cells. This tight coordination is also evident at the whole leaf level. Moreover, we observed an association of the properties of carbon-calcium inclusions and gas exchange, suggesting a possible implication of these structures in photosynthesis.

Reevaluation of the plant “gemstones”: Calcium oxalate crystals sustain photosynthesis under drought conditions

ABSTRACT Land plants face the perpetual dilemma of using atmospheric carbon dioxide for photosynthesis and losing water vapors, or saving water and reducing photosynthesis and thus growth. The reason behind this dilemma is that this simultaneous exchange of gases is accomplished through the same minute pores on leaf surfaces, called stomata. In a recent study we provided evidence that pigweed, an aggressive weed, attenuates this problem exploiting large crystals of calcium oxalate as dynamic carbon pools. This plant is able to photosynthesize even under drought conditions, when stomata are closed and water losses are limited, using carbon dioxide from crystal decomposition instead from the atmosphere. Abscisic acid, an alarm signal that causes stomatal closure seems to be implicated in this function and for this reason we named this path “alarm photosynthesis.” The so-far “enigmatic,” but highly conserved and widespread among plant species calcium oxalate crystals seem to play a crucial role in the survival of plants.