Seed Priming with Single-Walled Carbon Nanotubes Grafted with Pluronic P85 Preserves the Functional and Structural Characteristics of Pea Plants.

The engineering of carbon nanotubes in the last decades resulted in a variety of applications in electronics, electrochemistry, and biomedicine. A number of reports also evidenced their valuable application in agriculture as plant growth regulators and nanocarriers. In this work, we explored the effect of seed priming with single-walled carbon nanotubes grafted with Pluronic P85 polymer (denoted P85-SWCNT) on Pisum sativum (var. RAN-1) seed germination, early stages of plant development, leaf anatomy, and photosynthetic efficiency. We evaluated the observed effects in relation to hydro- (control) and P85-primed seeds. Our data clearly revealed that seed priming with P85-SWCNT is safe for the plant since it does not impair the seed germination, plant development, leaf anatomy, biomass, and photosynthetic activity, and even increases the amount of photochemically active photosystem II centers in a concentration-dependent manner. Only 300 mg/L concentration exerts an adverse effect on those parameters. The P85 polymer, however, was found to exhibit a number of negative effects on plant growth (i.e., root length, leaf anatomy, biomass accumulation and photoprotection capability), most probably related to the unfavorable interaction of P85 unimers with plant membranes. Our findings substantiate the future exploration and exploitation of P85-SWCNT as nanocarriers of specific substances promoting not only plant growth at optimal conditions but also better plant performance under a variety of environmental stresses.

Allelopathic Potential of the Cyanotoxins Microcystin-LR and Cylindrospermopsin on Green Algae.

Allelopathic interactions are widespread in all aquatic habitats, among all groups of aquatic primary biomass producers, including cyanobacteria. Cyanobacteria are producers of potent toxins called cyanotoxins, whose biological and ecological roles, including their allelopathic influence, are still incompletely understood. The allelopathic potential of the cyanotoxins microcystin-LR (MC-LR) and cylindrospermopsin (CYL) on green algae (Chlamydomonas asymmetrica, Dunaliella salina, and Scenedesmus obtusiusculus) was established. Time-dependent inhibitory effects on the growth and motility of the green algae exposed to cyanotoxins were detected. Changes in their morphology (cell shape, granulation of the cytoplasm, and loss of flagella) were also observed. The cyanotoxins MC-LR and CYL were found to affect photosynthesis to varying degrees in the green algae Chlamydomonas asymmetrica, Dunaliella salina, and Scenedesmus obtusiusculus, affecting chlorophyll fluorescence parameters such as the maximum photochemical activity (Fv/Fm) of photosystem II (PSII), the non-photochemical quenching of chlorophyll fluorescence (NPQ), and the quantum yield of the unregulated energy dissipation Y(NO) in PSII. In the context of ongoing climate change and the associated expectations of the increased frequency of cyanobacterial blooms and released cyanotoxins, our results demonstrated the possible allelopathic role of cyanotoxins on competing autotrophs in the phytoplankton communities.

Isoprene-Emitting Tobacco Plants Are Less Affected by Moderate Water Deficit under Future Climate Change Scenario and Show Adjustments of Stress-Related Proteins in Actual Climate.

Isoprene-emitting plants are better protected against thermal and oxidative stresses, which is a desirable trait in a climate-changing (drier and warmer) world. Here we compared the ecophysiological performances of transgenic isoprene-emitting and wild-type non-emitting tobacco plants during water stress and after re-watering in actual environmental conditions (400 ppm of CO2 and 28 °C of average daily temperature) and in a future climate scenario (600 ppm of CO2 and 32 °C of average daily temperature). Furthermore, we intended to complement the present knowledge on the mechanisms involved in isoprene-induced resistance to water deficit stress by examining the proteome of transgenic isoprene-emitting and wild-type non-emitting tobacco plants during water stress and after re-watering in actual climate. Isoprene emitters maintained higher photosynthesis and electron transport rates under moderate stress in future climate conditions. However, physiological resistance to water stress in the isoprene-emitting plants was not as marked as expected in actual climate conditions, perhaps because the stress developed rapidly. In actual climate, isoprene emission capacity affected the tobacco proteomic profile, in particular by upregulating proteins associated with stress protection. Our results strengthen the hypothesis that isoprene biosynthesis is related to metabolic changes at the gene and protein levels involved in the activation of general stress defensive mechanisms of plants.

Origin, evolution, and future of isoprene and nitric oxide interactions within leaves.

Photolytic generation of nitric oxide (NO), isoprene, and reactive oxygen species (ROS) pre-dated life on Earth (~4 billion years ago). However, isoprene-ROS-NO interactions became relevant to climate chemistry ~50 million years ago, after aquatic and terrestrial ecosystems became dominated by isoprene-emitting diatoms and angiosperms. Today, NO and NO2 (together referred to as NOx) are dangerous biogenic gaseous atmospheric pollutants. In plants, NO, with its multiple sources and sinks, acts as a secondary messenger that regulates development at low doses and induces cell death at high doses. Likewise, biogenic isoprene is a putative antioxidant and hormone 'enabler' that hastens plant (and leaf) growth and reproduction, and improves plant tolerance to transient abiotic stresses. Using examples from controlled-chamber simulation and field studies of isoprene oxidation, we discuss the likely nature and extent of isoprene oxidation within leaves. We argue that isoprene-NO interactions vary greatly among plant species, driven by differences in isoprene emission rate and nitrate assimilation capacity (i.e. NO sink strength), ROS availability, and the within-leaf ratio between free-NO and isoprene. In a warmer and CO2-fertilized future climate, antagonism between isoprene and NO within leaves will probably occur in a NO-rich (relative to present) environment, yielding a greater proportion of isoprene oxidation products, and inducing major changes in NO-mediated growth and stress responses.

Manipulation of light quality is an effective tool to regulate photosynthetic capacity and fruit antioxidant properties of Solanum lycopersicum L. cv. 'Microtom' in a controlled environment.

Light quality plays an essential role in setting plant structural and functional traits, including antioxidant compounds. This paper aimed to assess how manipulating the light spectrum during growth may regulate the photosynthetic activity and fruit bioactive compound synthesis in Solanum lycopersicum L. cv. 'Microtom' to improve plant physiological performance and fruit nutritional value. Plants were cultivated under three light quality regimes: red-green-blue LEDs (RGB), red-blue LEDs (RB) and white fluorescent lamps (FL), from sowing to fruit ripening. Leaf functional traits, photosynthetic efficiency, Rubisco and D1 protein expression, and antioxidant production in fruits were analyzed. Compared to FL, RGB and RB regimes reduced height and increased leaf number and specific leaf area, enhancing plant dwarf growth. The RGB regime improved photosynthesis and stomatal conductance despite lower biomass, favoring Rubisco synthesis and carboxylation rate than RB and FL regimes. The RB light produced plants with fewer flowers and fruits with a lower ascorbic acid amount but the highest polyphenol content, antioxidant capacity and SOD and CAT activities. Our data indicate that the high percentage of the green wavelength in the RGB regime promoted photosynthesis and reduced plant reproductive capacity compared to FL and RB. Conversely, the RB regime was the best in favoring the production of health-promoting compounds in tomato berries.

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.

Adaptive divergence generates distinct plastic responses in two closely related Senecio species.

The evolution of plastic responses to external cues allows species to maintain fitness in response to the environmental variations they regularly experience. However, it remains unclear how plasticity evolves during adaptation. To test whether distinct patterns of plasticity are associated with adaptive divergence, we quantified plasticity for two closely related but ecologically divergent Sicilian daisy species (Senecio, Asteraceae). We sampled 40 representative genotypes of each species from their native range on Mt. Etna and then reciprocally transplanted multiple clones of each genotype into four field sites along an elevational gradient that included the native elevational range of each species, and two intermediate elevations. At each elevation, we quantified survival and measured leaf traits that included investment (specific leaf area), morphology, chlorophyll fluorescence, pigment content, and gene expression. Traits and differentially expressed genes that changed with elevation in one species often showed little changes in the other species, or changed in the opposite direction. As evidence of adaptive divergence, both species performed better at their native site and better than the species from the other habitat. Adaptive divergence is, therefore, associated with the evolution of distinct plastic responses to environmental variation, despite these two species sharing a recent common ancestor.

Polymer-Modified Single-Walled Carbon Nanotubes Affect Photosystem II Photochemistry, Intersystem Electron Transport Carriers and Photosystem I End Acceptors in Pea Plants.

Single-walled carbon nanotubes (SWCNT) have recently been attracting the attention of plant biologists as a prospective tool for modulation of photosynthesis in higher plants. However, the exact mode of action of SWCNT on the photosynthetic electron transport chain remains unknown. In this work, we examined the effect of foliar application of polymer-grafted SWCNT on the donor side of photosystem II, the intersystem electron transfer chain and the acceptor side of photosystem I. Analysis of the induction curves of chlorophyll fluorescence via JIP test and construction of differential curves revealed that SWCNT concentrations up to 100 mg/L did not affect the photosynthetic electron transport chain. SWCNT concentration of 300 mg/L had no effect on the photosystem II donor side but provoked inactivation of photosystem II reaction centres and slowed down the reduction of the plastoquinone pool and the photosystem I end acceptors. Changes in the modulated reflection at 820 nm, too, indicated slower re-reduction of photosystem I reaction centres in SWCNT-treated leaves. We conclude that SWCNT are likely to be able to divert electrons from the photosynthetic electron transport chain at the level of photosystem I end acceptors and plastoquinone pool in vivo. Further research is needed to unequivocally prove if the observed effects are due to specific interaction between SWCNT and the photosynthetic apparatus.

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).

Single-Walled Carbon Nanotubes Modify Leaf Micromorphology, Chloroplast Ultrastructure and Photosynthetic Activity of Pea Plants.

Single-walled carbon nanotubes (SWCNTs) emerge as promising novel carbon-based nanoparticles for use in biomedicine, pharmacology and precision agriculture. They were shown to penetrate cell walls and membranes and to physically interact and exchange electrons with photosynthetic complexes in vitro. Here, for the first time, we studied the concentration-dependent effect of foliar application of copolymer-grafted SWCNTs on the structural and functional characteristics of intact pea plants. The lowest used concentration of 10 mg L-1 did not cause any harmful effects on the studied leaf characteristics, while abundant epicuticular wax generation on both leaf surfaces was observed after 300 mg L-1 treatment. Swelling of both the granal and the stromal regions of thylakoid membranes was detected after application of 100 mg L-1 and was most pronounced after 300 mg L-1. Higher SWCNT doses lead to impaired photosynthesis in terms of lower proton motive force generation, slower generation of non-photochemical quenching and reduced zeaxanthin content; however, the photosystem II function was largely preserved. Our results clearly indicate that SWCNTs affect the photosynthetic apparatus in a concentration-dependent manner. Low doses (10 mg L-1) of SWCNTs appear to be a safe suitable object for future development of nanocarriers for substances that are beneficial for plant growth.

The Interplay between Light Quality and Biostimulant Application Affects the Antioxidant Capacity and Photosynthetic Traits of Soybean (Glycine max L. Merrill).

This paper evaluates the combined effect of biostimulant and light quality on bioactive compound production and seedling growth of soybean (Glycine max L. Merrill) plants. Germinated seeds pre-treated with different concentrations (0.01%, 0.05%, 0.5%) of an amino acid-based biostimulant were grown for 4 days at the dark (D), white fluorescent light (FL), full-spectrum LED (FS), and red-blue (RB) light. Potential changes in the antioxidant content of sprouts were evaluated. Part of the sprouts was left to grow at FL, FS, and RB light regimes for 24 days to assess modifications in plants' anatomical and physiological traits during the early developmental plant stage. The seed pre-treatment with all biostimulant concentrations significantly increased sprout antioxidant compounds, sugar, and protein content compared to the control (seeds treated with H2O). The positive effect on bioactive compounds was improved under FS and RB compared to D and FL light regimes. At the seedling stage, 0.05% was the only concentration of biostimulant effective in increasing the specific leaf area (SLA) and photosynthetic efficiency. Compared to FL, the growth under FS and RB light regimes significantly enhanced the beneficial effect of 0.05% on SLA and photosynthesis. This concentration led to leaf thickness increase and shoot/root ratio reduction. Our findings demonstrated that seed pre-treatment with proper biostimulant concentration in combination with specific light regimes during plant development may represent a useful means to modify the bioactive compound amount and leaf structural and photosynthetic traits.

The effects of putrescine pre-treatment on osmotic stress responses in drought-tolerant and drought-sensitive wheat seedlings.

Recent studies have demonstrated that exogenous polyamines have protective effects under various stress condition. A broader understanding of the role of the polyamine pool fine regulation and the alterations of polyamine-related physiological processes could be obtained by comparing the stress effects in different genotypes. In this study, the impact of pre-treatment with putrescine in response to osmotic stress was investigated in the drought-tolerant Katya and drought-sensitive Zora wheat (Triticum aestivum) cultivars. Photosynthetic performance, in vivo thermoluminescence emission from leaves, leaf temperature, polyamine and salicylic acid levels, contents of osmoprotectants, and activities of antioxidant enzymes in the leaves were investigated not only to reveal differences in the physiological processes associated to drought tolerance, but to highlight the modulating strategies of polyamine metabolism between a drought-tolerant and a drought-sensitive wheat genotype. Results showed that the tolerance of Katya under osmotic stress conditions was characterized by higher photosynthetic ability, stable charge separation across the thylakoid membrane in photosystem II, higher proline accumulation and antioxidant activity. Thermoluminescence also revealed differences between the two varieties - a downshift of the B band and an increase of the afterglow band under osmotic stress in Zora, providing original complementary information to leaf photosynthesis. Katya variety exhibited higher constitutive levels of the signaling molecules putrescine and salicylic acid compared to the sensitive Zora. However, responses to exogenous putrescine were more advantageous for the sensitive variety under PEG treatment, which may be in relation with the decreased catabolism of polyamines, suggesting the increased need for polyamine under stress conditions.

Functional and Structural Leaf Plasticity Determine Photosynthetic Performances during Drought Stress and Recovery in Two Platanus orientalis Populations from Contrasting Habitats.

In the context of climatic change, more severe and long-lasting droughts will modify the fitness of plants, with potentially worse consequences on the relict trees. We have investigated the leaf phenotypic (anatomical, physiological and biochemical) plasticity in well-watered, drought-stressed and re-watered plants of two populations of Platanus orientalis, an endangered species in the west of the Mediterranean area. The two populations originated in contrasting climate (drier and warmer, Italy (IT) population; more humid and colder, Bulgaria (BG) population). The IT control plants had thicker leaves, enabling them to maintain higher leaf water content in the dry environment, and more spongy parenchyma, which could improve water conductivity of these plants and may result in easier CO2 diffusion than in BG plants. Control BG plants were also characterized by higher photorespiration and leaf antioxidants compared to IT plants. BG plants responded to drought with greater leaf thickness shrinkage. Drought also caused substantial reduction in photosynthetic parameters of both IT and BG plants. After re-watering, photosynthesis did not fully recover in either of the two populations. However, IT leaves became thicker, while photorespiration in BG plants further increased, perhaps indicating sustained activation of defensive mechanisms. Overall, our hypothesis, that plants with a fragmented habitat (i.e., the IT population) lose phenotypic plasticity but acquire traits allowing better resistance to the climate where they became adapted, remains confirmed.

Physiological and structural adjustments of two ecotypes of Platanus orientalis L. from different habitats in response to drought and re-watering.

Platanus orientalis covers a very fragmented area in Europe and, at the edge of its natural distribution, is considered a relic endangered species near extinction. In our study, it was hypothesized that individuals from the edge of the habitat, with stronger climate constrains (drier and warmer environment, Italy, IT ecotype), developed different mechanisms of adaptation than those growing under optimal conditions at the center of the habitat (more humid and colder environment, Bulgaria, BG ecotype). Indeed, the two P. orientalis ecotypes displayed physiological, structural and functional differences already under control (unstressed) conditions. Adaptation to a dry environment stimulated constitutive isoprene emission, determined active stomatal behavior, and modified chloroplast ultrastructure, ultimately allowing more effective use of absorbed light energy for photochemistry. When exposed to short-term acute drought stress, IT plants showed active stomatal control that enhanced instantaneous water use efficiency, and stimulation of isoprene emission that sustained photochemistry and reduced oxidative damages to membranes, as compared to BG plants. None of the P. orientalis ecotypes recovered completely from drought stress after re-watering, confirming the sensitivity of this mesophyte to drought. Nevertheless, the IT ecotype showed less damage and better stability at the level of chloroplast membrane parameters when compared to the BG ecotype, which we interpret as possible adaptation to hostile environments and improved capacity to cope with future, likely more recurrent, drought stress.

In Planta Recapitulation of Isoprene Synthase Evolution from Ocimene Synthases.

Isoprene is the most abundant biogenic volatile hydrocarbon compound naturally emitted by plants and plays a major role in atmospheric chemistry. It has been proposed that isoprene synthases (IspS) may readily evolve from other terpene synthases, but this hypothesis has not been experimentally investigated. We isolated and functionally validated in Arabidopsis the first isoprene synthase gene, AdoIspS, from a monocotyledonous species (Arundo donax L., Poaceae). Phylogenetic reconstruction indicates that AdoIspS and dicots isoprene synthases most likely originated by parallel evolution from TPS-b monoterpene synthases. Site-directed mutagenesis demonstrated invivo the functional and evolutionary relevance of the residues considered diagnostic for IspS function. One of these positions was identified by saturating mutagenesis as a major determinant of substrate specificity in AdoIspS able to cause invivo a dramatic change in total volatile emission from hemi- to monoterpenes and supporting evolution of isoprene synthases from ocimene synthases. The mechanism responsible for IspS neofunctionalization by active site size modulation by a single amino acid mutation demonstrated in this study might be general, as the very same amino acidic position is implicated in the parallel evolution of different short-chain terpene synthases from both angiosperms and gymnosperms. Based on these results, we present a model reconciling in a unified conceptual framework the apparently contrasting patterns previously observed for isoprene synthase evolution in plants. These results indicate that parallel evolution may be driven by relatively simple biophysical constraints, and illustrate the intimate molecular evolutionary links between the structural and functional bases of traits with global relevance.

Phenotypic differences determine drought stress responses in ecotypes of Arundo donax adapted to different environments.

Arundo donax has been identified as an important biomass and biofuel crop. Yet, there has been little research on photosynthetic and metabolic traits, which sustain the high productivity of A. donax under drought conditions. This study determined phenotypic differences between two A. donax ecotypes coming from stands with contrasting adaptation to dry climate. We hypothesized that the Bulgarian (BG) ecotype, adapted to drier conditions, exhibits greater drought tolerance than the Italian (IT) ecotype, adapted to a more mesic environment. Under well-watered conditions the BG ecotype was characterized by higher photosynthesis, mesophyll conductance, intrinsic water use efficiency, PSII efficiency, isoprene emission rate and carotenoids, whereas the IT ecotype showed higher levels of hydroxycinnamates. Photosynthesis of water-stressed plants was mainly limited by diffusional resistance to CO2 in BG, and by biochemistry in IT. Recovery of photosynthesis was more rapid and complete in BG than in IT, which may indicate better stability of the photosynthetic apparatus associated to enhanced induction of volatile and non-volatile isoprenoids and phenylpropanoid biosynthesis. This study shows that a large phenotypic plasticity among A. donax ecotypes exists, and may be exploited to compensate for the low genetic variability of this species when selecting plant productivity in constrained environments.

How Does the Amount and Composition of PM Deposited on Platanus acerifolia Leaves Change Across Different Cities in Europe?

Particulate matter (PM) deposited on Platanus acerifolia tree leaves has been sampled in the urban areas of 28 European cities, over 20 countries, with the aim of testing leaf deposited particles as indicator of atmospheric PM concentration and composition. Leaves have been collected close to streets characterized by heavy traffic and within urban parks. Leaf surface density, dimensions, and elemental composition of leaf deposited particles have been compared with leaf magnetic content, and discussed in connection with air quality data. The PM quantity and size were mainly dependent on the regional background concentration of particles, while the percentage of iron-based particles emerged as a clear marker of traffic-related pollution in most of the sites. This indicates that Platanus acerifolia is highly suitable to be used in atmospheric PM monitoring studies and that morphological and elemental characteristics of leaf deposited particles, joined with the leaf magnetic content, may successfully allow urban PM source apportionment.

Physiological significance of isoprenoids and phenylpropanoids in drought response of Arundinoideae species with contrasting habitats and metabolism.

Physiological, biochemical and morpho-anatomical traits that determine the phenotypic plasticity of plants under drought were tested in two Arundinoideae with contrasting habitats, growth traits and metabolism: the fast-growing Arundo donax, which also is a strong isoprene emitter, and the slow-growing Hakonechloa macra that does not invest on isoprene biosynthesis. In control conditions, A. donax displayed not only higher photosynthesis but also higher concentration of carotenoids and lower phenylpropanoid content than H. macra. In drought-stressed plants, photosynthesis was similarly inhibited in both species, but substantially recovered only in A. donax after rewatering. Decline of photochemical and biochemical parameters, increased concentration of CO2 inside leaves, and impairment of chloroplast ultrastructure were only observed in H. macra indicating damage of photosynthetic machinery under drought. It is suggested that volatile and non-volatile isoprenoids produced by A. donax efficiently preserve the chloroplasts from transient drought damage, while H. macra invests on phenylpropanoids that are less efficient in preserving photosynthesis but likely offer better antioxidant protection under prolonged stress.