Andrea Ertani

Effect of commercial lignosulfonate-humate on Zea mays L. metabolism.

Lignosulfonate-humate a and lignosulfonate-humate b, derived by an industrial process from lignin, were studied chemically and biologically, and their effects on maize metabolism compared with the responses induced by humic substances obtained from leonardite. Lignosulfonate-humate a and lignosulfonate-humate b elicited hormonelike activity and leonardite displayed giberellin properties. To improve our understanding of their biological action, lignosulfonate-humate a, lignosulfonate-humate b and leonardite were supplied to maize plants and their effect was studied on growth, nitrogen metabolism and photosynthesis. All products increased root and leaf growth. Glutamine-synthetase, glutamate-synthase enzyme activities and protein content were all increased. The treatments also increased chlorophyll content, glucose, fructose and rubisco enzyme activity, suggesting a positive role of lignosulfonate-humate a, lignosulfonate-humate b and leonardite in the photosynthetic process. In addition, an increase in phenol content was observed. In light of these results, being environmentally friendly products, lignosulfonate-humate a and lignosulfonate-humate b could be used to increase crop yield.

Effects of an alfalfa protein hydrolysate on the gene expression and activity of enzymes of the tricarboxylic acid (TCA) cycle and nitrogen metabolism in Zea mays L.

The effects through which an alfalfa protein hydrolysate (EM) possessing gibberellin- and auxin-like activity may promote plant nitrogen (N) nutrition have been investigated in Zea mays L. Treatment with 0.01 or 0.1 mg L(-1) EM for 48 h resulted in enhanced plant growth and leaf sugar accumulation. Concomitantly, the level of nitrates decreased, whereas total N percentage was unchanged. The activity of a number of enzymes involved in carbon (C) metabolism (malate dehydrogenase, MDH; isocitrate dehydrogenase, IDH; citrate synthase, CS) and N reduction and assimilation (nitrate reductase, NR; nitrite reductase, NiR; glutamine synthetase, GS; glutamate synthase, GOGAT; aspartate aminotransferase, AspAT) was significantly induced by EM supply to plants, and the transcription pattern of MDH, IDH, CS, and NR strongly correlated with data of enzyme activity. The transcript accumulation of asparagine synthetase (AS) was also induced by EM in the roots. The results suggest that EM might promote nitrogen assimilation in plants through a coordinate regulation of C and N metabolic pathways and open the way for further research on protein hydrolysates as a valid tool to improve N use efficiency and, as a consequence, to reduce the intensive use of inorganic N fertilizers in agriculture.

Plant biostimulants: physiological responses induced by protein hydrolyzed-based products and humic substances in plant metabolism

In recent years, the use of biostimulants in sustainable agriculture has been growing. Biostimulants can be obtained from different organic materials and include humic substances (HS), complex organic materials, beneficial chemical elements, peptides and amino acids, inorganic salts, seaweed extracts, chitin and chitosan derivatives, antitranspirants, amino acids and other N-containing substances. The application of biostimulants to plants leads to higher content of nutrients in their tissue and positive metabolic changes. For these reasons, the development of new biostimulants has become a focus of scientific interest. Among their different functions, biostimulants influence plant growth and nitrogen metabolism, especially because of their content in hormones and other signalling molecules. A significant increase in root hair length and density is often observed in plants treated with biostimulants, suggesting that these substances induce a “nutrient acquisition response” that favors nutrient uptake in plants via an increase in the absorptive surface area. Furthermore, biostimulants positively influence the activity and gene expression of enzymes functioning in the primary and secondary plant metabolism. This article reviews the current literature on two main classes of biostimulants: humic substances and protein-based biostimulants. The characteristic of these biostimulants and their effects on plants are thoroughly described.

Capsicum chinensis L. growth and nutraceutical properties are enhanced by biostimulants in a long-term period: chemical and metabolomic approaches

Two biostimulants, one derived from alfalfa plants (AH) and the other obtained from red grape (RG), were chemically characterized using enzyme linked immuno-sorbent assays, Fourier transform infrared (FT-IR) and Raman spectroscopies. Two doses (50 and 100 mL L−1 for RG, and 25 and 50 mL L−1 for AH) of biostimulants were applied to Capsicum chinensis L. plants cultivated in pots inside a tunnel. The experimental design consisted of the factorial combination of treatment (no biostimulant, plus AH, plus RG) at three doses (zero, low, and high) and two time-course applications (at the second and fourth week after transplantation) and the effects were recorded at flowering and maturity. Both biostimulants contained different amounts of indoleacetic acid and isopentenyladenosine; the AH spectra exhibited amino acid functional groups in the peptidic structure, while the RG spectra showed the presence of polyphenols, such as resveratrol. These results revealed that at flowering, RG and AH increased the weights of fresh leaves and fruits and the number of green fruits, whereas at maturity, the biostimulants most affected the fresh weight and number of red fruits. At flowering, the leaves of the treated plants contained high amounts of epicatechin, ascorbic acid, quercetin, and dihydrocapsaicin. At maturity, the leaves of the treated plants exhibited elevated amounts of fructose, glucose, chlorogenic, and ferulic acids. Moreover, green fruits exhibited a high content of chlorogenic acid, p-hydroxybenzoic acid, p-coumaric acid and antioxidant activity, while both AH- and RG-treated red fruits were highly endowed in capsaicin. The 1H high-resolution magic-angle spinning (HRMAS)-nuclear magnetic resonance (NMR) spectra of red fruits revealed that both products induced a high amount of NADP+, whereas RG also increased glucose, fumarate, ascorbate, thymidine and high molecular weight species. Our results suggested that AH and RG promoted plant growth and the production of secondary metabolites, such as phenols.

Spontaneous aggregation of humic acid observed with AFM at different pH

Atomic force microscopy in contact (AFM-C) mode was used to investigate the molecular dynamics of leonardite humic acid (HA) aggregate formed at different pH values. HA nanoparticles dispersed at pH values ranging from 2 to 12 were observed on a mica surface under dry conditions. The most clearly resolved and well-resulted AFM images of single particle were obtained at pH 5, where HA appeared as supramolecular particles with a conic shape and a hole in the centre. Those observations suggested that HA formed under these conditions exhibited a pseudo-amphiphilic nature, with secluded hydrophobic domains and polar subunits in direct contact with hydrophilic mica surface. Based on molecular simulation methods, a lignin-carbohydrate complex (LCC) model was proposed to explain the HA ring-like morphology. The LCC model optimized the parameters of β-O-4 linkages between 14 units of 1-4 phenyl propanoid, and resulted in an optimized structure comprising 45-50 linear helical molecules looped spirally around a central cavity. Those results added new insights on the adsorption mechanism of HA on polar surfaces as a function of pH, which was relevant from the point of view of natural aggregation in soil environment.

Chromium in Agricultural Soils and Crops: A Review

The mobility and distribution of metals in the environment is related not only to their concentration but also to their availability in the environment. Most chromium (Cr) exists in oxidation states ranging from 0 to VI in soils but the most stable and common forms are Cr(0), Cr(III), and Cr(VI) species. Chromium can have positive and negative effects on health, according to the dose, exposure time, and its oxidation state. The last is highly soluble; mobile; and toxic to humans, animals, and plants. On the contrary, Cr(III) has relatively low toxicity and mobility and it is one of the micronutrients needed by humans. In addition, Cr(III) can be absorbed on the surface of clay minerals in precipitates or complexes. Thus, the approaches converting Cr(VI) to Cr(III) in soils and waters have received considerable attention. The Cr(III) compounds are sparingly soluble in water and may be found in water bodies as soluble Cr(III) complexes, while the Cr(VI) compounds are readily soluble in water. Chromium is absorbed by plants through carriers of essential ions such as sulfate. Chromium uptake, accumulation, and translocation, depend on its speciation. Chromium shortage can cause cardiac problems, metabolic dysfunctions, and diabetes. Symptoms of Cr toxicity in plants comprise decrease of germination, reduction of growth, inhibition of enzymatic activities, impairment of photosynthesis and oxidative imbalances. This review provides an overview of the chemical characteristics of Cr, its behavior in the environment, the relationships with plants and aspects of the use of fertilizers.

Transcriptome-Wide Identification of Differentially Expressed Genes in Solanum lycopersicon L. in Response to an Alfalfa-Protein Hydrolysate Using Microarrays

An alfalfa-based protein hydrolysate (EM) has been tested in tomato (Solanum lycopersicon L.) plants at two different concentrations (0.1 and 1 mL L-1) to get insight on its efficacy as biostimulant in this species and to unravel possible metabolic targets and molecular mechanisms that may shed light on its mode of action. EM was efficient in promoting the fresh biomass and content in chlorophyll and soluble sugars of tomato plants, especially when it was applied at the concentration of 1 mL L-1. This effect on plant productivity was likely related to the EM-dependent up-regulation of genes identified via microarray and involved in primary carbon and nitrogen metabolism, photosynthesis, nutrient uptake and developmental processes. EM also up-regulated a number of genes implied in the secondary metabolism that leads to the synthesis of compounds (phenols and terpenes) functioning in plant development and interaction with the environment. Concomitantly, phenol content was enhanced in EM-treated plants. Several new genes have been identified in tomato as potential targets of EM action, like those involved in detoxification processes from reactive oxygen species and xenobiotic (particularly glutathione/ascorbate cycle-related and ABC transporters), and defense against abiotic and biotic stress. The model hypothesized is that elicitors present in the EM formulation like auxins, phenolics, and amino acids, may trigger a signal transduction pathway via modulation of the intracellular levels of the hormones ethylene, jasmonic acid and abscissic acid, which then further prompt the activation of a cascade events requiring the presence and activity of many kinases and transcription factors to activate stress-related genes. The genes identified suggest these kinases and transcription factors as players involved in a complex crosstalk between biotic and abiotic stress signaling pathways. We conclude that EM acts as a biostimulant in tomato due to its capacity to stimulate plant productivity and up-regulate stress-related responses. Its use in agricultural practices may reduce the need of inorganic fertilizers and pesticides, thereby reducing the environmental impact of productive agriculture.

Biological Activity of Vegetal Extracts Containing Phenols on Plant Metabolism.

The influence of vegetal extracts derived from red grape, blueberry fruits and hawthorn leaves on Zea mays L. plant growth and the activity of phenylalanine ammonia-lyase (PAL), a key enzyme of the phenylpropanoid pathway, was investigated in laboratory experiments. The extracts were characterized using FT-IR and Raman spectroscopies in order to obtain a pattern of the main functional groups. In addition, phenols content was determined by HPLC, whereas the content of indoleacetic acid and isopentenyladenosine hormones was determined by ELISA test and the auxin and gibberellin-like activities by plant-bioassays. The treated maize revealed increased root and leaf biomass, chlorophyll and sugars content with respect to untreated plants. Hawthorn, red grape skin and blueberry at 1.0 mL/L induced high p-coumaric content values, whilst hawthorn also showed high amounts of gallic and p-hydroxybenzoic acids. PAL activity induced by hawthorn at 1.0 mL/L had the highest values (11.1-fold UNT) and was strongly and linearly related with the sum of leaf phenols. Our results suggest that these vegetal extracts contain more than one group of plant-promoting substances.

Topsoil organic matter properties in contrasted hedgerow vegetation types

Background and aimsHedges, semi-natural landscape components, have the ability to integrate both agronomic and environmental functions and to provide several ecosystem services. The aim of this study was to test whether hedgerow vegetation is a determinant of soil organic matter properties in ancient agricultural lands.MethodsWe complemented cluster analysis and ordination to determine the extent to which two types of hedges that were distinct in character-plant species also differed between each other in concentration and composition of two major constituents of soil organic matter, namely humic substances and dissolved organic matter.ResultsThe two types of hedges were associated with significant differences in humic carbon content, hormone-like activity and molecular size of humic substances, which, in general, were more similar to those typical of forest than of agricultural soils. Moreover, we detected between-group differences in several phenolic acids.ConclusionsVariation of the topsoil biochemical properties of hedges may be explained by variation in their vegetation characteristics, similar to other ecosystems. Spontaneous vegetation in hedges perform an important role in controlling the variability of surface soil properties that influence the evolution of soil organic matter and nutrient availability in agricultural lands.

Evaluation of Seaweed Extracts From Laminaria and Ascophyllum nodosum spp. as Biostimulants in Zea mays L. Using a Combination of Chemical, Biochemical and Morphological Approaches

Seaweed extracts can be employed as biostimulants during crop cultivation owing to their positive effects on plant performance. Therefore, in this study one extract from Laminaria (A) and five extracts from Ascophyllum nodosum (B–F) were assayed on maize (Zea mays L.) plants supplied for 2 days with 0.5 mL L−1 of single products to evaluate their capacity to stimulate root growth and morphology, nutrition, and sugars accumulation. Firstly, extracts were chemically characterized via Fourier transform infrared (FT-IR) and FT-Raman spectroscopies, and their content in carbon, nitrogen, phenolic acids and hormones (indole-3-acetic acid, IAA, and Isopentenyladenosine, IPA) was quantified. The auxin like- and gibberellic acid -like activities of all extracts were also determined. FT-IR and FT-Raman spectra provided complementary information depicting distinct spectral pattern for each extract. Bands assigned to alginic and uronic acids were dominant in FT-IR spectra, while those corresponding to polyaromatic rings were evident in FT-Raman spectra. In general, extracts stimulated root growth, nutrition, esterase activity, and sugar content. However, they showed high variation in chemical features, which may explain their different capacity in triggering physiological responses in maize. Among A. nodosum extracts for instance, E was the most efficient in promoting root morphology traits, likely because of its elevate content in IAA (32.43 nM), while F extract was the highest in phenol content (1,933 mg L−1) and the most successful in improving plant nutrition. On the other hand, C extract was very effective in stimulating root elongation, but did not influence plant nutrition. B and D extracts induced similar positive effects on plants, although they greatly varied in chemical composition. Laminaria extract (A) differed from A. nodosum extracts, because of its low content in total phenols and the presence of both IAA- and GA-like activity. We conclude that all seaweed extracts acted as biostimulants in maize, but their chemical properties appeared crucial in predicting the physiological response preferentially elicited by individual seaweed extracts.