Cellulose Nanocrystals and Lignin Nanoparticles Extraction from Lemna minor L.: Acid Hydrolysis of Bleached and Ionic Liquid-Treated Biomass.

Using biomass to develop and obtain environmentally friendly and industrially applicable biomaterials is increasingly attracting global interest. Herein, cellulose nanocrystals (CNCs) and lignin nanoparticles (LNPs) were extracted from Lemna minor L., a freshwater free-floating aquatic species commonly called duckweed. To obtain CNCs and LNPs, two different procedures and biomass treatment processes based on bleaching or on the use of an ionic liquid composed of triethylammonium and sulfuric acid ([TEA][HSO4]), followed by acid hydrolysis, were carried out. Then, the effects of these treatments in terms of the thermal, morphological, and chemical properties of the CNCs and LNPs were assessed. The resulting nanostructured materials were characterized by using Fourier-transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD) spectroscopy, thermo-gravimetric analysis (TGA), and scanning electron microscopy (SEM). The results showed that the two methodologies applied resulted in both CNCs and LNPs. However, the bleaching-based treatment produced CNCs with a rod-like shape, length of 100-300 nm and width in the range of 10-30 nm, and higher purity than those obtained with ILs that were spherical in shape. In contrast, regarding lignin, IL made it possible to obtain spherical nanoparticles, as in the case of the other treatment, but they were characterized by higher purity and thermal stability. In conclusion, this research highlights the possibility of obtaining nanostructured biopolymers from an invasive aquatic species that is largely available in nature and how it is possible, by modifying experimental procedures, to obtain nanomaterials with different morphological, purity, and thermal resistance characteristics.

The effect of common duckweed (Lemna minor L.) extract on the shelf-life of beef burgers stored in modified atmosphere packs: A metabolomics approach.

The impact of duckweed extracts (DEs) on the shelf-life of packaged beef burgers was evaluated through classical assays and untargeted metabolomics. Beef burgers were formulated with an antioxidants-free control (CON), 1 g/kg sodium ascorbate (ASC), and increasing levels of a DEs, namely 1 (DE1), 5 (DE5), and 10 (DE10) g/kg, packaged under modified atmosphere and stored at 4 °C for 19 days. The DEs, abundant in phytochemicals, determined no issues with the hygienic status of the product. DEs modulated the redox status, being ineffective in preserving linolenic acid from peroxidation. However, the oxidation marker 2-nonenoic acid was down-accumulated in the DE10 sample following 19 days of storage, recording a lower glutathione:glutathione disulfide ratio. The accumulation of adipate semialdehyde revealed the inefficiency of DEs in coping with protein oxidation, while DEs prevented the accumulation of biogenic amines. Therefore, this work suggests a potential pro-oxidant role of the formulated DEs.

Biobased: Biostimulants and biogenic nanoparticles enter the scene.

Anthropogenic activities generally consume non-renewable resources and release polluting substances into the environment. Concerning agriculture, the cropping systems are almost based on exploiting non-renewable resources. In recent years, increasing interest has been devoted to reusing agricultural, food and other biomass wastes, considered relevant as they can be seen as resources rich in compounds that can find numerous applications. Biomass biorefining has been successfully applied and has opened up sustainable alternatives to the disposal of agricultural, agroindustrial and food wastes. In this area, an emerging, smart and environmentally friendly way to reduce the impact of waste on the environment is to obtain innovative materials for agriculture. Therefore, as part of biobased strategies, the use of waste biomass to obtain biostimulants and biogenic nanoparticles for crops has recently been proposed. Some research has shown that appropriate biostimulants and biobased nanomaterials have the potential to counteract some of the problems that plague agriculture. The above materials can improve crop performance, enhance crop tolerance to biotic and abiotic stresses, and improve plant nutrition. In light of the above, this review aspires, in the first part, to provide an overview of the ideas and central points that characterize the concept of bioeconomy and circular economy. In the second part, on the other hand, the most recent studies related to the valorization of various types of wastes leading to innovative biobased materials and their application in agriculture are presented. In particular, biostimulants and nanoparticles obtained through biogenic synthesis using agroindustrial and plant residues have been considered. In conclusion, the studies reported in this review show that the use of some biomasses to obtain the above materials represents a sustainable way of waste management and valorization, enabling innovative biobased materials for agriculture.

The Combination of Untargeted Metabolomics with Response Surface Methodology to Optimize the Functional Potential of Common Duckweed (Lemna minor L.).

The present study was designed to evaluate the functional potential of common duckweed (Lemna minor L.) as a source of bioactive compounds of nutraceutical interest. The untargeted profiling of the bioactive components of common duckweed was carried out through ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS), in parallel with assessing in vitro antioxidant and enzymatic inhibition properties. The optimization of extraction parameters was determined using the response surface methodology (RSM) through a 3-factor central composite design. The process parameters included extraction temperature, % of ethanol, and ultrasound power, while the response variables were the phenolic content (considering each main phenolic class), total glucosinolates, total carotenoids, the antioxidant potential, and enzyme inhibition activities. The results revealed that common duckweed was a rich source of carotenoids and total flavonoids (mainly flavones and flavonols), followed by phenolic acids, low-molecular-weight phenolics, and glucosinolates. Interestingly, the total flavones, total flavonols and total carotenoid equivalents showed the highest and most positive correlation values with the bioactive properties measured. Finally, the combined RSM approach and unsupervised statistics allowed us to point out the pivotal impact of ethanol percentage in the extraction solvent to recover the highest amounts of bioactive compounds efficiently.

Biochemical Insights into the Ability of Lemna minor L. Extract to Counteract Copper Toxicity in Maize.

Metal trace elements (MTE) can damage crops if present in excessive amounts in the environment. This research investigated the effect of a plant extract of an aquatic species, Lemna minor L. (duckweed) (LE), on the ability of maize to cope with copper (Cu) toxicity. LE reversed the effects of Cu2+ on photosynthetic activity (Pn), evapotranspiration (E), stomatal conductance (gs), sub-stomatal CO2 concentration (Ci) and biomass which did not differ from the untreated controls. LE did not regulate the amount of copper in maize leaves, but compared to Cu-treated samples, the extract decreased the hydrogen peroxide (H2O2; -26% on average) and malondialdehyde (MDA; -47% on average) content, regardless of the dosage applied. Furthermore, the activity of antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT) was significantly increased by LE compared to samples treated with Cu alone. Untargeted metabolomic profiling revealed that LE activated maize secondary metabolism, eliciting the content of non-enzymatic antioxidants (flavonoids, glutathione and glutathione-related compounds, tocopherols and tocotrienols) and modulating plant stress-related hormones (brassinosteroids and ABA derivatives). The results of this study are promising and pave the way for using duckweed as a biostimulant to trigger beneficial effects in maize and increase its resistance to MTEs.

Use of Biostimulants as a New Approach for the Improvement of Phytoremediation Performance-A Review.

Environmental pollution is one of the most pressing global issues, and it requires priority attention. Environmental remediation techniques have been developed over the years and can be applied to polluted sites, but they can have limited effectiveness and high energy consumption and costs. Bioremediation techniques, on the other hand, represent a promising alternative. Among them, phytoremediation is attracting particular attention, a green methodology that relies on the use of plant species to remediate contaminated sites or prevent the dispersion of xenobiotics into the environment. In this review, after a brief introduction focused on pollution and phytoremediation, the use of plant biostimulants (PBs) in the improvement of the remediation effectiveness is proposed. PBs are substances widely used in agriculture to raise crop production and resistance to various types of stress. Recent studies have also documented their ability to counteract the deleterious effects of pollutants on plants, thus increasing the phytoremediation efficiency of some species. The works published to date, reviewed and discussed in the present work, reveal promising prospects in the remediation of polluted environments, especially for heavy metals, when PBs derived from humic substances, protein and amino acid hydrolysate, inorganic salts, microbes, seaweed, plant extracts, and fungi are employed.

Synthesis of a Lignin/Zinc Oxide Hybrid Nanoparticles System and Its Application by Nano-Priming in Maize.

Nanotechnologies are attracting attention in various scientific fields for their technological and application potential, including their use as bio-activators and nanocarriers in agriculture. This work aimed to synthesize a hybrid material (ZnO@LNP) consisting of lignin nanoparticles containing zinc oxide (4 wt %). The synthesized ZnO hybrid material showed catalytic effect toward thermal degradation, as evidenced by the TGA investigation, while both spectroscopic and contact angle measurements confirmed a modification of surface hydrophilicity for the lignin nanoparticles due to the presence of hydrophobic zinc oxide. In addition, the antioxidant activity of the ZnO@LNP and the zinc release of this material were evaluated. At the application level, this study proposes for the first time the use of such a hybrid system to prime maize seeds by exploiting the release characteristics of this material. Concerning the dosage applied, ZnO@LNP promoted inductive effects on the early stages of seed development and plant growth and biomass development of young seedlings. In particular, the ZnO@LNP stimulated, in the primed seeds, a higher content of chlorophyll, carotenoids, anthocyanins, total phenols, and a better antioxidant activity, as supported by the lower levels of lipid peroxidation found when compared to the control samples.

Biogenic ZnO Nanoparticles Synthesized Using a Novel Plant Extract: Application to Enhance Physiological and Biochemical Traits in Maize.

The need to increase crop productivity and resistance directs interest in nanotechnology. Indeed, biogenic metal oxide nanoparticles can promote beneficial effects in plants, while their synthesis avoids the environmental impacts of conventional synthetic procedures. In this context, this research aimed to synthesize biogenic zinc oxide nanoparticles (ZnO-NPs) using, for the first time, an extract of a wild and spontaneous aquatic species, Lemna minor (duckweed). The effectiveness of this biogenic synthesis was evidenced for comparison with non-biogenic ZnO-NPs (obtained without using the plant extract), which have been synthesized in this research. XRD (X-ray diffraction), FE-SEM (field emission gun electron scanning microscopy), EDX (energy dispersive x-ray spectroscopy), TEM (transmission electron microscope) and UV-vis (ultraviolet-visible spectrophotometry) showed the biogenic approach effectiveness. The duckweed extract was subjected to UHPLC-ESI/QTOF-MS (ultra high-pressure liquid chromatography quadrupole time of flight mass spectrometry) phenolic profiling. This untargeted characterization highlighted a high and chemically diverse content in the duckweed extract of compounds potentially implicated in nanoparticulation. From an application standpoint, the effect of biogenic nanoparticles was investigated on some traits of maize subjected to seed priming with a wide range of biogenic ZnO-NPs concentrations. Inductive effects on the shoot and root biomass development were ascertained concerning the applied dosage. Furthermore, the biogenic ZnO-NPs stimulated the content of chlorophylls, carotenoids, and anthocyanin. Finally, the study of malondialdehyde content (MDA) as a marker of the oxidative status further highlighted the beneficial and positive action of the biogenic ZnO-NPs on maize.

Phytodepuration of Nitrate Contaminated Water Using Four Different Tree Species.

Water pollution by excessive amounts of nitrate (NO3-) has become a global issue. Technologies to clean up nitrate-contaminated water bodies include phytoremediation. In this context, this research aimed to evaluate four tree species (Salix alba L., Populus alba L., Corylus avellana L. and Sambucus nigra L.) to remediate nitrate-contaminated waters (100 and 300 mg L-1). Some physiological parameters showed that S. alba L. and P. alba L. increased particularly photosynthetic activity, chlorophyll content, dry weight, and transpired water, following the treatments with the above NO3- concentrations. Furthermore, these species were more efficient than the others studied in the phytodepuration of water contaminated by the two NO3- levels. In particular, within 15 days of treatment, S. alba L. and P. alba L. removed nitrate quantities ranging from 39 to 78%. Differently, C. avellana L. and S. nigra L. did not show particular responses regarding the physiological traits studied. Nonetheless, these species removed up to 30% of nitrate from water. In conclusion, these data provide exciting indications on the chance of using S. alba L. and P. alba L. to populate buffer strips to avoid NO3- environmental dispersion in agricultural areas.

Lignin Nanoparticles: A Promising Tool to Improve Maize Physiological, Biochemical, and Chemical Traits.

Lignin, and its derivatives, are the subject of current research for the exciting properties shown by this biomass. Particularly attractive are lignin nanoparticles for their eco- and biocompatibility compared to other nanomaterials. In this context, the effect of nanostructured lignin microparticles (LNP), obtained from alkaline lignin by acid treatment, on maize plants was investigated. To this end, maize seeds were primed with LNP at five concentrations: 80 mg L-1 (T80), 312 mg L-1 (T312), 1250 mg L-1 (T1250), 5000 mg L-1 (T5000) and 20,000 mg L-1 (T20000). Concerning the dose applied, LNP prompted positive effects on the first stages of maize development (germination and radicle length). Furthermore, the study of plant growth, biochemical and chemical parameters on the developed plants indicated that concerning the dose applied. LNP stimulated beneficial effects on the seedlings (fresh weight and length of shoots and roots). Besides, specific treatments increased the content of chlorophyll (a and b), carotenoid, and anthocyanin. Finally, the soluble protein content showed a positive trend in response to specific dosages. These effects are significant, given the essential biological function performed by these biomolecules. In conclusion, this research indicates as the nanostructured lignin microparticles can be used, at appropriate dosages, to induce positive biological responses in maize. This beneficial action deserves attention as it candidates LNP for biostimulating a crop through seed priming.

Can biostimulants be used to mitigate the effect of anthropogenic climate change on agriculture? It is time to respond.

Anthropogenic climate change, namely climate alterations induced by human activities, is causing some issues to agricultural systems for their vulnerability to extreme events. Forecasts predict a global population increase in the near years that will exacerbate this situation, elevating the global demand for food. It will pose severe concerns in terms of natural resource usage and availability. Agriculture is one of the anthropogenic activities that will be more affected in the future. Climate extremes menace to affect the quantity and quality of crop production severely. Drought, water and soil salinity are considered among the most problematic factors that anthropogenic climate change will increase. This complex and worrying scenario requires the urgent implementation of sustainable measures which are capable of improving crop yield and quality, fostering the robustness and resilience of cropping systems. Among the more current methodology, the use of natural plant biostimulants (PBs) has been proposed to improve plant resistance to abiotic environmental stresses. The advantage of using these substances is due to their effectiveness in improving crop productivity and quality. Therefore, in this review, the most recent researches dealing with the use of natural PBs for improving plant resistance to drought and salinity, in an anthropogenic climate change scenario, have been reported and critically discussed.

Blue:Red LED Light Proportion Affects Vegetative Parameters, Pigment Content, and Oxidative Status of Einkorn (Triticum monococcum L. ssp. monococcum) Wheatgrass.

This work aimed to study the effect of some light spectra on the growth, oxidative state, and stress of einkorn wheatgrass (Triticum monococcum L. ssp. monococcum). To this end, six light treatments, having the same total incident photon flux density (PFD) of 200 µmol m-2 s-1, were applied to einkorn and compared: only blue light; only red; three blue:red combinations, at different proportions of total PFD (75:25%, 50:50%, and 25:75%, respectively); and a wide spectrum, taken as a control treatment, composed of blue (18% of PFD), red (18%), and intermediate wavelengths (64%). Light treatments affected the contents of pigments (chlorophylls and carotenes), hydrogen peroxide (H2O2), and malondialdehyde (MDA). These results revealed the changes in the oxidative status of wheatgrass, in response to the different light treatments. However, the dichromatic light with blue ≥50% of the total PFD appeared to be the best combination, guarantying good wheatgrass yield, increasing pigment content, and reducing H2O2 and MDA when compared to the other light treatments. Our findings also contribute to explaining the available literature on the effect of these kinds of light on the increase in phenolic compounds and antioxidant activity in einkorn wheatgrass.

Phytoremediation and detoxification of xenobiotics in plants: herbicide-safeners as a tool to improve plant efficiency in the remediation of polluted environments. A mini-review.

Phytoremediation is a widely studied and applied technology, based on the use of plants and their associated microorganisms to decontaminate polluted sites. In recent years, different strategies have been investigated to improve the phytoremediation efficiency of the selected plants. In this context, some studies have shown that herbicide-safeners, chemicals applied to crops to enhance their tolerance to herbicides, can increase the phytoremediation of soils and water polluted by organic and inorganic contaminants. Safeners, by inducing the xenobiotic detoxification and the antioxidant metabolism in plants, can enhance their removal potential in the cleaning process. In this review, after a short survey of phytoremediation technologies and the biochemical mechanisms activated by plants to tolerate and detoxify heavy metals and herbicides, the use of herbicide-safeners as a tool to increase the phytoremediation performance is reviewed and discussed.

Effects of Megafol on the Olive Cultivar 'Arbequina' Grown Under Severe Saline Stress in Terms of Physiological Traits, Oxidative Stress, Antioxidant Defenses, and Cytosolic Ca2.

Salinity is one of the most impacting abiotic stresses regarding crop productivity and quality. Among the strategies that are attracting attention in the protection of crops from abiotic stresses, there is the use of plant biostimulants. In this study, Megafol (Meg), a commercial plant biostimulant, was tested on olive plants subjected to severe saline stress. Plants treated with salt alone showed substantial reductions in biomass production, leaf net photosynthesis (Pn), leaf transpiration rate (E), stomatal conductance (gs), and relative water content (RWC). In addition, samples stressed with NaCl showed a higher sodium (Na+) content in the leaves, while those stressed with NaCl and biostimulated with Meg increased the potassium (K+) content in the leaves, thus showing a higher K+/Na+ ratio. Salinity caused the accumulation of significant quantities of hydrogen peroxide (H2O2) and malondialdehyde (MDA) due to decreases in the activity of antioxidant enzymes, namely superoxide dismutase (SOD - EC 1.15.1.1), ascorbate peroxidase (APX - EC 1.11.1.11), guaiacol peroxidase (GPX - EC 1.11.1.9), and catalase (CAT - EC 1.11.1.6). When olive plants under saline stress were biostimulated with Meg, the plants recovered and showed physiological and biochemical traits much improved than salt stressed samples. Finally, Meg exhibited Ca2+-chelating activity in olive pollen grains, which allowed the biostimulant to exert this beneficial effect also by antagonizing the undesirable effects of hydrogen peroxide on Ca2+ metabolism.

Application of a Plant Biostimulant To Improve Maize (Zea mays) Tolerance to Metolachlor.

Plant biostimulants (PBS) increase crop productivity and induce beneficial processes in plants. Although PBS can stimulate plant tolerance to some abiotic stresses, their effect in improving crop resistance to herbicide injuries has barely been investigated. Therefore, a study on the effect of a biostimulant (Megafol) on maize (Zea mays L.) tolerance to a chloro-acetanilide herbicide (metolachlor) was carried out. We found that Megafol reduced the negative effects of metolachlor on maize. Indeed, biostimulated samples showed increases in germination, biomass production, Vigor index, and EC50 (effective concentration causing 50% reductions to roots and aerial biomass) with respect to the samples treated with metolachlor alone. Furthermore, plants treated with the herbicide in combination with Megafol showed lower levels of malondialdehyde (MDA). Antioxidant enzymes, namely, ascorbate peroxidase (APX), guaiacol peroxidase (GPX), and catalase (CAT), were assayed in samples treated with metolachlor alone or in combination with Megafol, and higher enzymes activities were found in biostimulated plants. The results of this study open the perspective of using Megafol, as well as other suitable plant biostimulants, in improving the crop's capacity to cope with injuries and unwanted effects that herbicide could cause to these species.

The treatment of duckweed with a plant biostimulant or a safener improves the plant capacity to clean water polluted by terbuthylazine.

Water pollution is becoming alarming since thousands contaminants are dispersed in the aquatic environments, and agricultural practices, for the massive use of pesticides, are contributing to exacerbating this problem. In this context, a research aimed at investigating the ability of duckweed (Lemna minor), a free-floating aquatic species widespread throughout the world, to remediate water polluted with five different concentrations of a herbicide - terbuthylazine (TBA) - was carried out. In addition, duckweed was treated with a plant biostimulant and a safener with the aim of increasing the plant's capacity to tolerate and remove the TBA from the water. The results evidenced that the herbicide affected the duckweed already at the lower concentrations, reducing its capacity to proliferate and the area of its fronds. On the contrary, when the TBA treatments were performed in combination with the biostimulant or the safener the average area of the fronds was affected of lesser extents, compared to the plants treated with the herbicide only. Antioxidant enzymes, namely ascorbate peroxidases (APX) and catalases (CAT), were investigated and it was found that the biostimulated and safened duckweed showed increased activities of these enzymes, compared to the plants treated with TBA only. At last, some phytofiltration experiments were planned. The biostimulated and safened duckweed removed more TBA from polluted water than the plants treated with the herbicide alone. In conclusion, this research showed that duckweed is suitable for cleaning water polluted with TBA and this potential can be successfully improved by treating the species with a biostimulant or a safener.

Interference of three herbicides on iron acquisition in maize plants.

The use of herbicides to control weed species could lead to environmental threats due to their persistence and accumulation in the ecosystems and cultivated fields. Nonetheless, the effect of these compounds on plant mineral nutrition in crops has been barely investigated. This study aimed at ascertaining the effect of three herbicides (S-metolachlor, metribuzin and terbuthylazine) on the capacity of maize to acquire iron (Fe). Interferences on plant growth and reductions on the Fe contents were found in the plants treated. Furthermore, root cell viability and functionality losses were ascertained following the treatments, which, in turn, decreased the amount of phytosiderophores (PSs) released by the roots. An investigation carried out in greater depth on root apices of treated plants using an FE-SEM (Scanning Electron Microscope) coupled with EDX (Energy Dispersive X-ray) indicated that the reductions on Fe content started in this part of the roots. Lastly, decreases were found also in copper (Cu+2), zinc (Zn+2) and manganese (Mn+2) content in root apices.

Combination of aquatic species and safeners improves the remediation of copper polluted water.

In the last decades, many anthropogenic activities have resulted in heavy metal contamination of freshwaters and surrounding environments. This poses serious threats to human health. Phytoremediation is a cost-effective technology which is useful for remediating polluted soils and water. Recently, the use of aquatic free-floating plants has been proposed to remediate polluted water. In this context, a study on the capacity of two aquatic plants, Lemna minor (duckweed) and Salvinia auriculata (salvinia), to remediate Cu+2 (Cu) polluted water was carried out. Initially, the species were exposed to different copper concentrations (1, 5, 10, 20 and 50µmolL-1) in order to assess Cu+2 toxicity to the plants. In addition, plants were treated with two safeners (benoxacor and dichlormid), with the aim of pointing out any safening effect of these compounds on the aquatic species. Toxicity tests showed that safened plants had a greater Cu resistance, especially at the higher Cu doses. Finally, unsafened and safened plants were tested in the decontamination of water polluted by copper (1.2mgL-1). In general, duckweed removed higher amounts of Cu from polluted water than salvinia, and, surprisingly, for both the species the safeners significantly increased the plants' capacity to remove the metal from the polluted waters. Lastly, an HPLC-based method was developed and standardized to monitor the residual amounts of the two safeners in the water. While dichlormid was completely absorbed by duckweed within few days after the treatments, some residual amounts of both safeners were found in salvinia vegetated water after two weeks. In conclusion, the results of this research show that the use of aquatic species in combination with safeners is an attractive and reliable tool to make plants more effective in phytoremediation of water polluted with metals (or other toxic compounds).

Effect of three safeners on sulfur assimilation and iron deficiency response in barley (Hordeum vulgare) plants.

BACKGROUND: Safeners are agrochemicals used in agriculture to protect crops from herbicide injuries. They act by stimulating herbicide metabolism. As graminaceous plants, to cope with iron (Fe) deficiency, activate sulfur (S) metabolism and release huge amounts of Fe-chelating compounds, or phytosiderophores (PSs), we investigated, in barley plants (Hordeum vulgare, L.) grown in Fe deficiency, the effects of three safeners on two enzymes of S assimilation, cysteine (Cys) and glutathione (GSH), and PS release. Finally, we monitored the root Fe content in plants treated with the most effective safener. RESULTS: Generally, all the safeners activated S metabolism and increased Cys and GSH contents. In addition, the safened plants excreted higher levels of PSs. Given that mefenpyr-diethyl (Mef) was the most effective in causing these effects, we assessed the Fe concentration in Mef-treated barley and found higher Fe levels than those in untreated plants. CONCLUSION: The three safeners, in different ways but specifically, activated S reductive metabolism and regulated Cys and GSH contents, PS release rate and Fe content (Mef-treated barley). The results of this research provide new indications of the biochemical and physiological mechanisms involved in the safening action. © 2016 Society of Chemical Industry.