Photosynthetic Efficiency of Marchantia polymorpha L. in Response to Copper, Iron, and Zinc.

Metal micronutrients are essential for plant nutrition, but their toxicity threshold is low. In-depth studies on the response of light-dependent reactions of photosynthesis to metal micronutrients are needed, and the analysis of chlorophyll a fluorescence transients is a suitable technique. The liverwort Marchantia polymorpha L., a model organism also used in biomonitoring, allowed us to accurately study the effects of metal micronutrients in vivo, particularly the early responses. Gametophytes were treated with copper (Cu), iron (Fe) or zinc (Zn) for up to 120 h. Copper showed the strongest effects, negatively affecting almost the entire light phase of photosynthesis. Iron was detrimental to the flux of energy around photosystem II (PSII), while the acceptor side of PSI was unaltered. The impact of Fe was milder than that of Cu and in both cases the structures of the photosynthetic apparatus that resisted the treatments were still able to operate efficiently. The susceptibility of M. polymorpha to Zn was low: although the metal affected a large part of the electron transport chain, its effects were modest and short-lived. Our results may provide a contribution towards achieving a more comprehensive understanding of response mechanisms to metals and their evolution in plants, and may be useful for supporting the development of biomonitoring techniques.

Selenium Biofortification Impacts the Tomato Fruit Metabolome and Transcriptional Profile at Ripening.

In the present work, the effects of enriching tomatoes with selenium were studied in terms of physiological, metabolic, and molecular processes in the last stages of fruit development, particularly during ripening. A selenium concentration of 10 mg L-1 with sodium selenate and selenium nanoparticles was used in the spray treatments on the whole plants. No significant effects of selenium enrichment were detected in terms of ethylene production or color changes in the ripening fruit. However, selenium enrichment had an influence on both the primary and secondary metabolic processes and thus the biochemical composition of ripe tomatoes. Selenium decreased the amount of ß-carotene, increased the accumulation of naringenin and chlorogenic acid, and decreased the coumaric acid level. Selenium also affected the volatile organic compound profile, with changes in the level of specific apocarotenoid compounds, such as ß-ionone. These metabolomic changes may, to some extent, be due to the impact of selenium treatment on the transcription of genes involved in the metabolism of these compounds. RNA-seq analysis showed that the selenium application mostly impacted the expression of the genes involved in hormonal signaling, secondary metabolism, flavonoid biosynthesis, and glycosaminoglycan degradation.

Early evidence of the impacts of microplastic and nanoplastic pollution on the growth and physiology of the seagrass Cymodocea nodosa.

Microplastics (MPs) and nanoplastics (NPs) are ubiquitous in natural habitats and the risks their presence poses to marine environments and organisms are of increasing concern. There is evidence that seagrass meadows are particularly prone to accumulate plastic debris, including polystyrene particles, but the impacts of this pollutant on seagrass performance are currently unknown. This is a relevant knowledge gap as seagrasses provide multiple ecosystem services and are declining globally due to anthropogenic impact and climate-change-related stressors. Here, we explored the potential effects of a 12 day-exposure of seagrasses to one concentration (68 µg/L) of polystyrene MPs and NPs on the growth, oxidative status, and photosynthetic efficiency of plants using the foundation species Cymodocea nodosa as a model. Among plant organs, adventitious roots were particularly affected by MPs and NPs showing complete degeneration. The number of leaves per shoot was lower in MPs- and NPs-treated plants compared to control plants, and leaf loss exceeded new leaf production in MPs-treated plants. MPs also reduced photochemical efficiency and increased pigment content compared to control plants. Shoots of NPs-treated plants showed a greater oxidative damage and phenol content than those of control plants and MPs-treated plants. Biochemical data about oxidative stress markers were consistent with histochemical results. The effects of MPs on C. nodosa could be related to their adhesion to plant surface while those of NPs to entering tissues. Our study provides the first experimental evidence of the potential harmful effects of MPs/NPs on seagrass development. It also suggests that the exposure of seagrasses to MPs/NPs in natural environments could have negative consequences on the functioning of seagrass ecosystems. This stresses the importance of implementing cleaning programs to remove all plastics already present in marine habitats as well as of undertaking specific actions to prevent the introduction of these pollutants within seagrass meadows.

Effect of Zinc Priming on Salt Response of Wheat Seedlings: Relieving or Worsening?

In an attempt to alleviate salt-induced damage, the application of ZnO nanoparticles has been suggested. As the use of these particles has also been associated with phytotoxicity, to better clarify the effect of zinc and its possible mitigation of salt stress, we treated wheat seedlings with ZnO (nanoparticles or their bulk-scale counterparts, amended either in the growth medium, NPs and B, or sprayed on the leaves, SPNPs and SPB) with or without subsequent treatment with salt. Growth, photosynthetic parameters, zinc and ion concentration, and in situ and biochemical determination of oxidative stress in wheat leaves and/or in roots were considered. Both Zn and NaCl significantly inhibited growth and induced severe alterations in root morphology. Oxidative stress and damage decreased or increased under ZnO treatment and in saline conditions depending on the organ and on the size and mode of application of particles. In spite of the higher stress conditions often recorded in treated leaves, neither pigment concentration nor photochemical efficiency were decreased. A large variability in the effects of ZnO treatment/priming on seedling salt response was recorded; however, the presence of a cumulative negative effect of priming and salt stress sometimes observed calls for caution in the use of ZnO in protection from saline stress.

The Moss Leptodictyum riparium Counteracts Severe Cadmium Stress by Activation of Glutathione Transferase and Phytochelatin Synthase, but Slightly by Phytochelatins.

In the present work, we investigated the response to Cd in Leptodictyum riparium, a cosmopolitan moss (Bryophyta) that can accumulate higher amounts of metals than other plants, even angiosperms, with absence or slight apparent damage. High-performance liquid chromatography followed by electrospray ionization tandem mass spectrometry of extracts from L. riparium gametophytes, exposed to 0, 36 and 360 µM Cd for 7 days, revealed the presence of γ-glutamylcysteine (γ-EC), reduced glutathione (GSH), and traces of phytochelatins. The increase in Cd concentrations progressively augmented reactive oxygen species levels, with activation of both antioxidant (catalase and superoxide dismutase) and detoxifying (glutathione-S-transferase) enzymes. After Cd treatment, cytosolic and vacuolar localization of thiol peptides was performed by means of the fluorescent dye monochlorobimane and subsequent observation with confocal laser scanning microscopy. The cytosolic fluorescence observed with the highest Cd concentrations was also consistent with the formation of γ-EC-bimane in the cytosol, possibly catalyzed by the peptidase activity of the L. riparium phytochelatin synthase. On the whole, activation of phytochelatin synthase and glutathione-S-transferase, but minimally phytochelatin synthesis, play a role to counteract Cd toxicity in L. riparium, in this manner minimizing the cellular damage caused by the metal. This study strengthens previous investigations on the L. riparium ability to efficiently hinder metal pollution, hinting at a potential use for biomonitoring and phytoremediation purposes.

TiO2 nanoparticles may alleviate cadmium toxicity in co-treatment experiments on the model hydrophyte Azolla filiculoides.

The hydrophyte Azolla filiculoides can be a useful model to assess if TiO2 NPs may in some way alleviate the Cd injuries and improve the ability of the plant to cope with this metal. With this mechanistic hypothesis, after a pre-treatment with TiO2 NPs, A. filiculoides plants were transferred to cadmium-contaminated water with or without TiO2 nanoparticles. After 5 days of treatment, cadmium uptake, morpho-anatomical, and physiological aspects were studied in plants. The continuous presence of TiO2 nanoparticles, though not increasing the uptake of cadmium in comparison with a priming treatment, induced a higher translocation of this heavy metal to the aerial portion. Despite the translocation factor was always well below 1, cadmium contents in the fronds, generally greater than 100 ppm, ranked A. filiculoides as a good cadmium accumulator. Higher cadmium contents in leaves did not induce damages to the photosynthetic machinery, probably thanks to a compartmentalization strategy aimed at confining most of this pollutant to less metabolically active peripheral cells. The permanence of NPs in growth medium ensured a better efficiency of the antioxidant apparatus (proline and glutathione peroxidase and catalase activities) and induced a decrease in H2O2 content, but did not suppress TBARS level.

Modulation of the defence responses against Cd in willow species through a multifaceted analysis.

Willow, due to the extensive root system, high transpiration rates and ability to accumulate large amounts of cadmium, is considered particularly useful for green remediation practices. In this study two different willow species, Salix viminalis and Salix alba, were used to assess possible differences in their ability of cadmium accumulation and to analyse in detail the physiology of their response to treatments with this metal using a multidisciplinary approach. Plants were grown in hydroponics and treated with 0, 50 and 100 µM Cd2+ (CdCl2) for 7 and 14 days. Cadmium content, oxidative stress, both evaluated by biochemical and histochemical techniques, antioxidant response, leaf stomatal conductance and photosynthetic efficiency were measured in control and treated roots and/or leaves. The two willow species removed cadmium with a high efficiency from the growth solution; however, the highest contents of Cd recorded in plants grown in the presence of the lower Cd concentrations suggest a limited capacity of metal accumulation. No photochemical limitation characterised treated plants, probably due to the ability to store large amounts of Cd in the root compartment, with reduction of damage to the photosynthetic machinery. S. viminalis, able to uptake cadmium also in the root apical region, seemed to be a more efficient accumulator than S. alba and, thanks to a relatively higher antioxidant response, did not show a higher level of oxidative stress. On the basis of the above, the two plant species, in particular S. viminalis, are confirmed as useful for cadmium phytostabilisation/phytoextraction.

Soyasaponins from Zolfino bean as aldose reductase differential inhibitors.

Seven triterpenoid saponins were identified in methanolic extracts of seeds of the Zolfino bean landrace (Phaseolus vulgaris L.) by HPLC fractionation, revealing their ability to inhibit highly purified human recombinant aldose reductase (hAKR1B1). Six of these compounds were associated by MS analysis with the following saponins already reported in different Phaseolus vulgaris varieties: soyasaponin Ba (V), soyasaponin Bb, soyasaponin Bd (sandosaponin A), soyasaponin αg, 3-O-[R-l-rhamnopyranosyl(1 → 2)-α-d-glucopyranosyl(1 → 2)-α-d-glucuronopyranosyl]olean-12-en-22-oxo-3α,-24-diol, and soyasaponin ßg. The inhibitory activity of the collected fractions containing the above compounds was tested for hAKR1B1-dependent reduction of both l-idose and 4-hydroxynonenal, revealing that some are able to differentially inhibit the enzyme. The present work also highlights the difficulties in the search for aldose reductase differential inhibitors (ARDIs) in mixtures due to the masking effect on ARDIs exerted by the presence of conventional aldose reductase inhibitors. The possibility of differential inhibition generated by a different inhibitory model of action of molecules on different substrates undergoing transformation is also discussed.

Can sugar metabolism in the cambial region explain the water deficit tolerance in poplar?

Drought dramatically affects wood production by adversely impacting cambial cells and their derivatives. Photosynthesis and assimilate transport are also affected by drought conditions. Two poplar genotypes, Populus deltoides 'Dvina' and Populus alba 'Marte', demonstrated contrasting growth performance and water-carbon balance strategies; a mechanistic understanding of the water deficit response was provided by these poplar species. 'Marte' was found to be more anisohydric than 'Dvina'. This characteristic was associated with the capacity to reallocate carbohydrates during water deficits. In contrast, 'Dvina' displayed more conservative water management; carbohydrates were preferably stored or used for cellulose production rather than to achieve an osmotic balance between the phloem and the xylem. Data confirmed that the more 'risk-taking' characteristic of 'Marte' allowed a rapid recovery following water deficit and was connected to a different carbohydrate metabolism.

Growth and physiological features of Chaetomorpha linum (Müller) Kütz. in high density mats.

Seasonal changes of high density Chaetomorpha linum mats were studied in an eutrophic coastal lagoon. We measured biomass and specific growth rate and analyzed water quality, sediment labile organic matter (LOM), photosynthetic efficiency and pigments in two areas: one subjected to sediment resuspension and algal mass shuffling through specific boats and the other left undisturbed. Low disturbance enhanced algal survival under critical summer conditions, by reducing LOM and promoting growth through thalli fragmentation. Chaetomorpha linum grew fast and quickly acclimated its physiology to adverse conditions, although the mat underlayer showed low photosynthetic efficiency. Nevertheless, the ability to respond to the adverse environment was not sufficient to prevent the sudden algal decay, which occurred following a prolonged summer stress. Present results may help to plan the lagoon management through boat operations, to attain a more effective control over the algal growth and a more efficient removal of the mat.

Edible vegetables as a source of aldose reductase differential inhibitors.

The hyperactivity of aldose reductase (AR) on glucose in diabetic conditions or on glutathionyl-hydroxynonenal in oxidative stress conditions, the source of cell damage and inflammation, appear to be balanced by the detoxifying action exerted by the enzyme. This detoxification acts on cytotoxic hydrophobic aldehydes deriving from membrane peroxidative processes. This may contribute to the failure in drug development for humans to favorably intervene in diabetic complications and inflammation, despite the specificity and high efficiency of several available aldose reductase inhibitors. This paper presents additional features to a previously proposed approach, on inhibiting the enzyme through molecules able to preferentially inhibit the enzyme depending on the substrate the enzyme is working on. These differential inhibitors (ARDIs) should act on glucose reduction catalyzed by AR without little or no effect on the reduction of alkenals or alkanals. The reasons why AR may be an eligible enzyme for differential inhibition are considered. These mainly refer to the evidence that, although AR is an unspecific enzyme that recognizes different substrates such as aldoses and hydrophobic aldehydes, it nevertheless displays a certain degree of specificity among substrates of the same class. After screening on edible vegetables, indications of the presence of molecules potentially acting as ARDIs are reported.

Zolfino landrace (Phaseolus vulgaris L.) from Pratomagno: general and specific features of a functional food.

BACKGROUND: The Zolfino bean is a variety of Phaseolus vulgaris, which is cultivated in a limited area of Tuscany, Italy, and is widely appreciated for its flavor and culinary uses. OBJECTIVES: A yellow Zolfino landrace cultivated in the Leccio-Reggello area was characterized and compared with three other varieties of Phaseolus vulgaris (i.e. the Borlotto, Cannellino, and Corona beans) in terms of its general features and potential as an antioxidant/anti-inflammatory agent. DESIGN: The length, width, thickness, equatorial section surface, weight, volume, and seed coat section were measured in all the beans. The seed surface area was also estimated by an original empirical method. The ability of the different beans to interfere with the enzymes of the polyol pathway (that is, aldose reductase (AR) and sorbitol dehydrogenase) was tested using the supernatant after soaking the beans at room temperature and after thermal treatment, which simulated the bean-cooking process in a controlled fashion. RESULTS: Concerning the general features, Zolfino was comparable with other beans, except Corona, in terms of surface-volume ratio, which possesses the lowest tegument thickness. Moreover, Zolfino appears the most effective in inhibiting AR activity. The inhibitory ability is unaffected by thermal treatment and appears to be associated with compound(s) present in the coat of the bean. CONCLUSIONS: The ability of Zolfino to inhibit AR, thus reducing the flux of glucose through the polyol pathway, highlights the features of Zolfino as a functional food, potentially useful in treating the dysfunctions linked to the hyperactivity of AR, such as diabetic complications or inflammatory responses.

A new approach to macroalgal bloom control in eutrophic, shallow-water, coastal areas.

In summer 2012, an experiment was conducted in a shallow eutrophic lagoon with poor water exchange to determine the consequences of harvesting algae on the algal mat itself, which was traversed and repeatedly disturbed by large harvester boats. Four areas with high macroalgal density, measuring half a hectare each, were selected. Two were subjected to frequent disturbance of the algal mat and sediment (12 two-hour operations over a 38-day period) and the other two were left undisturbed as control. The following variables were determined: 1) water column physical chemistry and nutrients; 2) redox potential, nutrients and organic load in sediments; 3) C, N and P content of algal thalli; 4) macroalgal biomass. In 2013, a further experiment was conducted on a larger scale. Biomass was estimated in a high-density mat measuring 235 ha, where macroalgae were harvested and stirred up by four harvesting boats, and in two high-density mats measuring 150 and 120 ha, left undisturbed as control (9.15, 9.92 and 3.68 kg/m(2), respectively). In the first experiment, no significant changes were observed in the water column. In sediment the main variation was a significant reduction in labile organic matter in the disturbed areas and a significant increase mainly in refractory organic matter in the undisturbed areas. Biomass showed a significant drastic reduction in disturbed areas and substantial stability in undisturbed areas. In the large-scale experiment, the biomass of the disturbed mat declined by about 63%, only 6.5% of which was due to harvesting. On the other hand, the undisturbed mat with higher density underwent a natural decline in biomass of about 23% and the other increased by about 50%. These results demonstrate that disturbance of high-density mat in shallow water by boats can cause decay of the mat.

Hormonal signals involved in the regulation of cambial activity, xylogenesis and vessel patterning in trees.

The radial growth of plant stem is based on the development of cribro-vascular cambium tissues. It affects the transport efficiency of water, mineral nutrients and photoassimilates and, ultimately, also plant height. The rate of cambial cell divisions for the assembly of new xylem and phloem tissue primordia and the rate of differentiation of the primordia into mature tissues determine the amount of biomass produced and, in the case of woody species, the wood quality. These complex physiological processes proceed at a rate which depends on several factors, acting at various levels: growth regulators, resource availability and environmental factors. Several hormonal signals and, more recently, further regulatory molecules, have been shown to be involved in the induction and maintenance of cambium and the formation of secondary vascular tissues. The control of xylem cell patterning is of particular interest, because it determines the diameter of xylem vessels, which is central to the efficiency of water and nutrient transport from roots to leaves through the stem and may strongly influence the growth in height of the tree. Increasing scientific evidence have proved the role of other hormones in cambial cell activities and the study of the hormonal signals and their crosstalking in cambial cells may foster our understanding of the dynamics of xylogenesis and of the mechanism of vessel size control along the stem. In this article, the role of the hormonal signals involved in the control of cambium and xylem development in trees and their crosstalking are reviewed.

Nitric oxide and hydrogen peroxide involvement during programmed cell death of Sechium edule nucellus.

The nucellus is a maternal tissue that feeds the developing embryo and the secondary endosperm. During seed development the cells of the nucellus suffer a degenerative process early after fertilization as the cellular endosperm expands and accumulates reserves. Nucellar cell degeneration has been characterized as a form of developmentally programmed cell death (PCD). In this work we show that nucellus PCD is accompanied by a considerable production of both nitric oxide and hydrogen peroxide (NO and H(2)O(2)). Interestingly, each of the two molecules is able to induce the production of the other and to cause cell death when applied to a living nucellus. We show that the induced cell death has features of a PCD, accompanied by profound changes in the morphology of the nuclei and by a massive degradation of nuclear DNA. Moreover, we report that NO and H(2)O(2) cause an induction of caspase-like proteases previously characterized in physiological nucellar PCD.

Indoleacetic acid concentration and metabolism changes during bud development in tubers of two potato (Solanum tuberosum) cultivars.

Plant growth regulators are involved in the control of potato (Solanum tuberosum) tuber dormancy. Evidence concerning the role of IAA is controversial; we therefore investigated its role by analyzing two cultivars with varying lengths of dormancy. We examined the time course of free and conjugated IAA in tuber tissue isolates from the final stages of tuber growth to the end of dormancy, the distribution of free IAA in tuber tissues by in situ analysis, and the biosynthesis of the hormone by feeding experiments. The time course of free IAA showed marked differences between the examined cultivars, although the concentration of the auxin generally was the highest at the early stages of tuber dormancy. Immunodetection showed a similar pattern of IAA distribution in both genotypes: in dormant buds from freshly harvested tubers, the free hormone accumulated mostly in apical meristem, leaf and lateral bud primordia, and differentiating vascular tissues underlying the apical meristem, while at the end of the storage period only axillary bud primordia from growing buds displayed appreciable auxin levels. Feeding experiments indicated that changes in IAA biosynthesis rate were a major cause of auxin variation in buds. In both cultivars, dormancy apparently ceased when free IAA fell below a threshold value. Despite this, our data led us to conclude that IAA would not be directly responsible for inhibiting sprouting. Instead, auxin might shorten dormancy, in a cultivar-dependent manner, by enhancing early developmental processes in buds, ultimately leading to dormancy termination.

The involvement of indole-3-acetic acid in the control of stem elongation in dark- and light-grown pea (Pisum sativum) seedlings.

We investigated the role of auxin on stem elongation in pea (Pisum sativum L.) grown for 10d in continuous darkness or under low-irradiance blue, red, far red and white light. The third internode of treated seedlings was peeled and the tissues (epidermis and cortex+central cylinder) were separately analyzed for the concentration of free and conjugated indole-3-acetic acid (IAA). Under red, far red and white light internode elongation was linearly related with the free IAA content of all internode tissues, suggesting that phytochrome-dependent inhibition of stem growth may be mediated by a decrease of free IAA levels in pea seedlings. The correlation between IAA and internode elongation, however, did not hold for blue light-grown seedlings. The hypothesis that the growth response under low-irradiance blue light might be correlated with the lack of phytochrome B signalling and changes in gibberellin metabolism is discussed in view of current knowledge on hormonal control of stem growth.

Secretory proteins as potential semiochemical carriers in the horse.

Two soluble proteins were isolated as major secretory products of horse sweat and of the parotid gland and characterized for structural and functional properties. The first protein, lipocalin allergen EquC1, was characterized for its glycosylation sites and bound glycosidic moieties. Only one (Asn53) of the two putative glycosylation sites within the sequence was post-translationally modified; a different glycosylation pattern was determined with respect to data previously reported. When purified from horse sweat, this protein contained oleamide and other organic molecules as natural ligands. Ligand binding experiments indicated good protein selectivity toward volatile compounds having a straight chain structure of 9-11 carbon atoms, suggesting a role of this lipocalin in chemical communication. The second protein, here reported for the first time in the horse, belongs to the group of parotid secretory proteins, part of a large superfamily of binding proteins whose function in most cases is still unclear. This protein was sequenced and characterized for its post-translational modifications. Of the three cysteine residues present, two were involved in a disulfide bridge (Cys155-Cys198). A model, built up on the basis of similar proteins, indicated a general fold characterized by the presence of a long hydrophobic barrel. Binding experiments performed with a number of different organic compounds failed to identify ligands for this protein with a well-defined physiological role.

Are xylem radial development and hydraulic conductivity in downwardly-growing grapevine shoots influenced by perturbed auxin metabolism?

Downwardly-growing grapevine shoots have smaller and more frequent vessels than upwardly-growing ones and, as a consequence, a lower hydraulic conductivity. Here, grapevine (Vitis vinifera L.) shoot growth orientation was manipulated to test whether downward shoot orientation negatively affects vessel growth in the apex via a shortage of water and nutrients. The orientation of the central vine shoot portion was inverted by two consecutive 135 degrees bends, resulting in double-bent N-shaped vines; the central downward shoot portion was of different lengths in the experimental treatments to induce increasing reductions of shoot conductivity. These treatments reduced shoot conductivity and water flow, but had no effects on vessel development and frequency in the apex. In a second experiment, auxin concentration was assessed in shoots of upwardly- and downwardly-growing plants. IAA concentration at the apical internodes was higher in downwardly-oriented shoots than in shoots growing upwards. In addition, a higher density and a lower vessel diameter were observed in the lower, than the upper side, of the downwardly-oriented shoot, suggesting increased accumulation of auxin in the lower side. These results suggest that the downward orientation induces accumulation of auxin in the apex, which in turn affects the density and the size of the xylem vessels, causing reduction of hydraulic conductivity.