Effects of Two Doses of Organic Extract-Based Biostimulant on Greenhouse Lettuce Grown Under Increasing NaCl Concentrations.
ABSTRACT: The enhancement of plant tolerance toward abiotic stresses is increasingly being supported by the application of biostimulants. Salinity represents a serious problem in the Mediterranean region. To verify the effects deriving from the application of biostimulants, trials on Romaine lettuce plants under salt exposure were performed, in greenhouse. Plants were subjected to three NaCl solutions with 0.8, 1.3, and 1.8 dS/m of electrical conductivity. The volume of the solution was 200 mL/plant and delivered every 3 days. Biostimulant treatments started after crop establishment and were: control (water) and two doses (0.1 or 0.2 mL/plant) of the commercial biostimulant Retrosal® (Valagro S.p.A), containing calcium, zinc, and specific active ingredients. Four Retrosal® treatments were applied, every 7 days, directly to the substrate. Non-destructive analyses were conducted to assess the effects on leaf photosynthetic efficiency. At harvest, plants fresh weight (FW) and dry weight were determined, as well as the concentration of chlorophylls, carotenoids, total sugars, nitrate, proline, and abscisic acid (ABA). The biostimulant tested increased significantly the FW of lettuce (+65% in the highest dose) compared to controls. Results indicate that treatments positively affected the chlorophyll content measured in vivo (+45% in the highest dose) and that a general positive effect was observable on net photosynthesis rate. Retrosal® seems to improve the gas exchanges under our experimental conditions. The total sugars levels were not affected by treatments. Biostimulant allowed maintaining nitrate concentration similar to the untreated and unstressed controls. The increasing levels of water salinity caused a raise in proline concentration in control plants (+85%); biostimulant treatments at 0.2 mL/plant dose kept lower the proline levels. All plants treated with the biostimulant showed lower value of ABA (-34%) compared to controls. Results revealed that Retrosal® is able to stimulate plant growth independently from the salinity exposure. However, treated plants reached faster the commercial maturity stage. The fresh biomass of control at the end of experiment, after 30 days, ranged from 15 to 42 g/head, while in biostimulant treated plants ranged from 45 to 94 g/head. The product applied at maximum dose seems to be the most effective in our experimental conditions.
Project description:Water deficit causes substantial yield losses that climate change is going to make even more problematic. Sustainable agricultural practices are increasingly developed to improve plant tolerance to abiotic stresses. One innovative solution amongst others is the integration of plant biostimulants in agriculture. In this work, we investigate for the first time the effects of the biostimulant -Leafamine<sup>®</sup>-a protein hydrolysate on greenhouse lettuce <i>(Lactuca sativa</i> L.) grown under well-watered and water-deficit conditions. We examined the physiological and metabolomic water deficit responses of lettuce treated with Leafamine<sup>®</sup> (0.585 g/pot) or not. Root application of Leafamine<sup>®</sup> increased the shoot fresh biomass of both well-watered (+40%) and deficit-irrigated (+20%) lettuce plants because the projected leaf area increased. Our results also indicate that Leafamine<sup>®</sup> application could adjust the nitrogen metabolism by enhancing the total nitrogen content, amino acid (proline) contents and the total protein level in lettuce leaves, irrespective of the water condition. Osmolytes such as soluble sugars and polyols, also increased in Leafamine<sup>®</sup>-treated lettuce. Our findings suggest that the protective effect of Leafamine is a widespread change in plant metabolism and could involve ABA, putrescine and raffinose.
Project description:Modern agriculture is facing many difficulties due to a rapidly changing climate, and environmental damage from agricultural production. The commitment of scientists and farmers to increase environmentally sustainable agricultural practices is one way to help mitigate environmental impacts. Among these practices, the use of biostimulants could be beneficial for increasing fertilizer efficiency and reducing excessive use in agriculture, and as plant growth regulators capable of increasing both production volume and quality of crops. In our study, rocket plants were grown in a greenhouse and treated with two biostimulants (protein hydrolysates or tropical plant extract), either individually or combined, to assess the effect on yield, dry biomass, mineral content, qualitative parameters as well as on economic profitability of foliar biostimulant applications. Total yield and dry biomass of the plants treated with the three biostimulant combinations on average increased by 48.1% and 37.2% respectively compared to untreated plants, without significant differences between treatments. Biostimulant application increased the content of chlorophyll, K, Mg and Ca, compared to the untreated plants. Furthermore, a biostimulant synergistic effect was detected concerning the content of total ascorbic acid. Our results confirmed that the biostimulants are eco-friendly products, able to boost plant growth and product quality and thus increase growers' profitability.
Project description:Microbial inoculants such as <i>Trichoderma</i>-based products are receiving great interest among researchers and agricultural producers for their potential to improve crop productivity, nutritional quality as well as resistance to plant pathogens/pests and numerous environmental stresses. Two greenhouse experiments were conducted to assess the effects of <i>Trichoderma</i>-based biostimulants under suboptimal, optimal and supraoptimal levels of nitrogen (N) fertilization in two leafy vegetables: Iceberg lettuce (<i>Lactuca sativa</i> L.) and rocket (<i>Eruca sativa</i> Mill.). The yield, nutritional characteristics, N uptake and mineral composition were analyzed for each vegetable crop after inoculation with <i>Trichoderma</i> strains <i>T. virens</i> (GV41) or <i>T. harzianum</i> (T22), and results were compared to non-inoculated plants. In addition, the effect of the <i>Trichoderma</i>-based biostimulants on microbes associated with the rhizosphere in terms of prokaryotic and eukaryotic composition and concentration using DGGE was also evaluated. <i>Trichoderma</i>-based biostimulants, in particular GV41, positively increased lettuce and rocket yield in the unfertilized plots. The highest marketable lettuce fresh yield was recorded with either of the biostimulant inoculations when plants were supplied with optimal levels of N. The inoculation of rocket with GV41, and to a lesser degree with T22, elicited an increase in total ascorbic acid under both optimal and high N conditions. <i>T. virens</i> GV41 increased N-use efficiency of lettuce, and favored the uptake of native N present in the soil of both lettuce and rocket. The positive effect of biostimulants on nutrient uptake and crop growth was species-dependent, being more marked with lettuce. The best biostimulation effects from the <i>Trichoderma</i> treatments were observed in both crops when grown under low N availability. The <i>Trichoderma</i> inoculation strongly influenced the composition of eukaryotic populations in the rhizosphere, in particularly exerting different effects with low N levels in comparison to the N fertilized plots. Overall, inoculations with <i>Trichoderma</i> may be considered as a viable strategy to manage the nutrient content of leafy horticulture crops cultivated in low fertility soils, and assist vegetable growers in reducing the use of synthetic fertilizers, developing sustainable management practices to optimize N use efficiency.
Project description:Microbial diversity is suggested as the key for plant and human health. However, how microbial diversity can be enriched is largely unknown but of great interest for health issues. Biostimulants offer the way to directly augment our main living areas by the healthy microbiome of indoor plants. Here, we investigated shifts of the microbiome on leaves of spider plants (Chlorophytum comosum) and its surrounding abiotic surfaces in the built environment after irrigation with a vermicompost-based biostimulant for 12 weeks. The biostimulant could not only promote plant growth, but changed the composition of the microbiome and abundance of intact microbial cells on plant leaves and even stronger on abiotic surfaces in close vicinity under constant conditions of the microclimate. Biostimulant treatments stabilized microbial diversity and resulted in an increase of Bacteroidetes and a surprising transient emerge of new phyla, e.g., Verrucomicrobia, Acidobacteria, and Thaumarchaeota. The proportion of potentially beneficial microorganisms like Brevibacillus, Actinoallomurus, Paenibacillus, Sphaerisporangium increased relatively; microbial diversity was stabilized, and the built environment became more plant-like. Detected metabolites like indole-3-acetic acid in the biostimulant were potentially contributed by species of Pseudomonas. Overall, effects of the biostimulant on the composition of the microbiome could be predicted with an accuracy of 87%. This study shows the potential of biostimulants not only for the plant itself, but also for other living holobionts like humans in the surrounding environment.
Project description:Environmental stresses have a significant effect on agricultural crop productivity worldwide. Exposure of seeds to abiotic stresses, such as salinity among others, results in lower seed viability, reduced germination, and poor seedling establishment. Alternative agronomic practices, e.g., the use of plant biostimulants, have attracted considerable interest from the scientific community and commercial enterprises. Biostimulants, i.e., products of biological origin (including bacteria, fungi, seaweeds, higher plants, or animals) have significant potential for (i) improving physiological processes in plants and (ii) stimulating germination, growth and stress tolerance. However, biostimulants are diverse, and can range from single compounds to complex matrices with different groups of bioactive components that have only been partly characterized. Due to the complex mixtures of biologically active compounds present in biostimulants, efficient methods for characterizing their potential mode of action are needed. In this study, we report the development of a novel complex approach to biological activity testing, based on multi-trait high-throughput screening (MTHTS) of <i>Arabidopsis</i> characteristics. These include the <i>in vitro</i> germination rate, early seedling establishment capacity, growth capacity under stress and stress response. The method is suitable for identifying new biostimulants and characterizing their mode of action. Representatives of compatible solutes such as amino acids and polyamines known to be present in many of the biostimulant irrespective of their origin, i.e., well-established biostimulants that enhance stress tolerance and crop productivity, were used for the assay optimization and validation. The selected compounds were applied through seed priming over a broad concentration range and the effect was investigated simultaneously under control, moderate stress and severe salt stress conditions. The new MTHTS approach represents a powerful tool in the field of biostimulant research and development and offers direct classification of the biostimulants mode of action into three categories: (1) plant growth promotors/inhibitors, (2) stress alleviators, and (3) combined action.
Project description:Microbial plant biostimulants have been successfully applied to improve plant growth, stress resilience and productivity. However, the mechanisms of action of biostimulants are still enigmatic, which is the main bottleneck for the fully realization and implementation of biostimulants into the agricultural industry. Here, we report the elucidation of a global metabolic landscape of maize (<i>Zea mays</i> L) leaves in response to a microbial biostimulant, under well-watered and drought conditions. The study reveals that the increased pool of tricarboxylic acid (TCA) intermediates, alterations in amino acid levels and differential changes in phenolics and lipids are key metabolic signatures induced by the application of the microbial-based biostimulant. These reconfigurations of metabolism gravitate toward growth-promotion and defense preconditioning of the plant. Furthermore, the application of microbial biostimulant conferred enhanced drought resilience to maize plants <i>via</i> altering key metabolic pathways involved in drought resistance mechanisms such as the redox homeostasis, strengthening of the plant cell wall, osmoregulation, energy production and membrane remodeling. For the first time, we show key molecular events, metabolic reprogramming, activated by a microbial biostimulant for plant growth promotion and defense priming. Thus, these elucidated metabolomic insights contribute to ongoing efforts in decoding modes of action of biostimulants and generating fundamental scientific knowledgebase that is necessary for the development of the plant biostimulants industry, for sustainable food security.
Project description:This review presents a comprehensive and systematic study of the field of plant biostimulants and considers the fundamental and innovative principles underlying this technology. The elucidation of the biological basis of biostimulant function is a prerequisite for the development of science-based biostimulant industry and sound regulations governing these compounds. The task of defining the biological basis of biostimulants as a class of compounds, however, is made more complex by the diverse sources of biostimulants present in the market, which include bacteria, fungi, seaweeds, higher plants, animals and humate-containing raw materials, and the wide diversity of industrial processes utilized in their preparation. To distinguish biostimulants from the existing legislative product categories we propose the following definition of a biostimulant as "a formulated product of biological origin that improves plant productivity as a consequence of the novel or emergent properties of the complex of constituents, and not as a sole consequence of the presence of known essential plant nutrients, plant growth regulators, or plant protective compounds." The definition provided here is important as it emphasizes the principle that biological function can be positively modulated through application of molecules, or mixtures of molecules, for which an explicit mode of action has not been defined. Given the difficulty in determining a "mode of action" for a biostimulant, and recognizing the need for the market in biostimulants to attain legitimacy, we suggest that the focus of biostimulant research and validation should be upon proof of efficacy and safety and the determination of a broad mechanism of action, without a requirement for the determination of a specific mode of action. While there is a clear commercial imperative to rationalize biostimulants as a discrete class of products, there is also a compelling biological case for the science-based development of, and experimentation with biostimulants in the expectation that this may lead to the identification of novel biological molecules and phenomenon, pathways and processes, that would not have been discovered if the category of biostimulants did not exist, or was not considered legitimate.
Project description:The ongoing unpredictability of climate changes is exponentially exerting a negative impact on crop production, further aggravating detrimental abiotic stress effects. Several research studies have been focused on the genetic modification of crop plants to achieve more crop resilience against such stress factors; however, there has been a paradigm shift in modern agriculture focusing on more organic, eco-friendly and long-lasting systems to improve crop yield. As such, extensive research into the use of microbial and nonmicrobial biostimulants has been at the core of agricultural studies to improve crop growth and development, as well as to attain tolerance against several biotic and abiotic stresses. However, the molecular mechanisms underlying the biostimulant activity remain enigmatic. Thus, this study is a liquid chromatography-mass spectrometry (LC-MS)-based untargeted metabolomics approach to unravel the hypothetical biochemical framework underlying effects of a nonmicrobial biostimulant (a silicon-based formulation) on tomato plants (<i>Solanum lycopersium</i>) under salinity stress conditions. This metabolomics study postulates that Si-based biostimulants could alleviate salinity stress in tomato plants through modulation of the primary metabolism involving changes in the tricarboxylic acid cycle, fatty acid and numerous amino acid biosynthesis pathways, with further reprogramming of several secondary metabolism pathways such as the phenylpropanoid pathway, flavonoid biosynthesis pathways including flavone and flavanol biosynthesis. Thus, the postulated hypothetical framework, describing biostimulant-induced metabolic events in tomato plants, provides actionable knowledge necessary for industries and farmers to, confidently and innovatively, explore, design, and fully implement Si-based formulations and strategies into agronomic practices for sustainable agriculture and food production.
Project description:The aim of this study was to examine the transcriptional changes that govern the primary nitrogen (N) limitation response in roots and the long-term effect of N limitation in the basal nodes, which may be associated with tiller suppression by N supply. The effect of biostimulant application on N limitation response was also examined by including plant treated and untreated with microalgae extract-based biostimulant. For this purpose, RNA-seq was conducted in roots of 15 days-old hydroponically grown wheat plants (treated and untreated with biostimulants) 24 h after the introduction of the plants to the N limitation and in basal nodes of 32 days-old wheat plants (treated and untreated with biostimulants) exposed to N limitation for 18 days. The two different N treatments were: High N (10 mM N) and Low N (0.1 mM N). In total, three biological replicates were included per treatment. RNA-seq was conducted in total RNA extracted from 3 basal node samples pooled together per biological replicate. In this study, the basal node is defined as the 0.5 cm of the main shoot base, which includes the apical meristem, lateral buds, leaf meristems etc.
Project description:Humic substance (HS)-based biostimulants show potentials as sustainable strategies for improved crop development and stress resilience. However, cellular and molecular mechanisms governing the agronomically observed effects of HS on plants remain enigmatic. Here, we report a global metabolic reprogramming of maize leaves induced by a humic biostimulant under normal and nutrient starvation conditions. This reconfiguration of the maize metabolism spanned chemical constellations, as revealed by molecular networking approaches. Plant growth and development under normal conditions were characterized by key differential metabolic changes such as increased levels of amino acids, oxylipins and the tricarboxylic acid (TCA) intermediate, isocitric acid. Furthermore, under starvation, the humic biostimulant significantly impacted pathways that are involved in stress-alleviating mechanisms such as redox homeostasis, strengthening of the plant cell wall, osmoregulation, energy production and membrane remodelling. Thus, this study reveals that the humic biostimulant induces a remodelling of inter-compartmental metabolic networks in maize, subsequently readjusting the plant physiology towards growth promotion and stress alleviation. Such insights contribute to ongoing efforts in elucidating modes of action of biostimulants, generating fundamental scientific knowledge that is necessary for development of the biostimulant industry, for sustainable food security.