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Microalgae are a promising feedstock with proven biostimulant activity that is enhanced by their biochemical components (e.g., amino acids and phytohormones), which turns them into an appealing feedstock to reduce the use of fertilisers in agriculture and improve crop productivity and resilience. Thus, this work aimed to isolate protein-rich microalgal mutants with increased biostimulant activity. Random mutagenesis was performed with Chlorella vulgaris, and a selection of protein-rich mutants were sorted through fluorescence-activated cell sorting (FACS), resulting in the isolation of 17 protein-rich mutant strains with protein contents 19-34% higher than that of the wildtype (WT). Furthermore, mutant F4 displayed a 38%, 22% and 62% higher biomass productivity, growth rate and chlorophyll content, respectively. This mutant was then scaled up to a 7 L benchtop reactor to produce biomass and evaluate the biostimulant potential of this novel strain towards garden cress seeds. Compared to water (control), the germination index and the relative total growth increased by 7% and 19%, respectively, after the application of 0.1 g L-1 of this bioproduct, which highlights its biostimulant potential.

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BACKGROUND: Cauliflower (Brassica oleracea L.) and globe artichoke (Cynara scolymus L.) are vegetables with a high waste index mainly related to stems and leaves. In this study, enzymatic hydrolysates obtained from these wastes were proposed to be used as plant biostimulants. Life cycle assessment methodology was also applied to evaluate environmental performances related to cauliflower and artichoke byproducts. RESULTS: Hydrolysates (HYs) were chemically and biologically characterized. Amino acids, organic acids, amines, polyols, mineral elements, phenols, tannins, flavonoids and sulfur compounds were identified and quantified by means of NMR, inductively coupled plasma mass spectrometry and UV-visible analyses. Cauliflower leaf and flower HYs showed the highest concentration of free amino acids, whereas stems showed the highest concentration of Ca. Regarding artichoke, asparagine, glutamine and aspartic acid were exclusively detected in stems, whereas artichoke leaves showed the highest Mg and Mn levels together with the highest antioxidant activity. The HYs diluted in water were tested as biostimulants. The impacts of five concentrations of HYs (0.00, 0.28, 0.84, 2.52 and 7.56 g L-1) on seed germination and early seedling growth of crimson clover, alfalfa, durum wheat and corn were investigated. CONCLUSIONS: The application of artichoke biostimulant (0.28 g L-1) positively influenced the coefficient of velocity of germination in alfalfa, crimson clover and durum wheat, whereas cauliflower biostimulant significantly improved corn germination speed. © 2024 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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The best environment for plant growth and development contains certain essential metabolites. A broad category of metabolites known as "plant biostimulants" (PBs) includes biomolecules such as proteins, carbohydrates, lipids, and other secondary metabolites related to groups of terpenes, specific nitrogen-containing compounds, and benzene ring-conjugated compounds. The formation of biomolecules depends on both biotic and abiotic factors, such as the release of PB by plants, animals, and microorganisms, or it can result from the control of temperature, humidity, and pressure in the atmosphere, in the case of humic substances (HSs). Understanding the genomic outputs of the concerned organism (may be plants or others than them) becomes crucial for identifying the underlying behaviors that lead to the synthesis of these complex compounds. For the purposes of achieving the objectives of sustainable agriculture, detailed research on PBs is essential because they aid in increasing yield and other growth patterns of agro-economic crops. The regulation of homeostasis in the plant-soil-microbe system for the survival of humans and other animals is mediated by the action of plant biostimulants, as considered essential for the growth of plants. The genomic size and gene operons for functional and regulation control have so far been revealed through technological implementations, but important gene annotations are still lacking, causing a delay in revealing the information. Next-generation sequencing techniques, such as nanopore, nanoball, and Illumina, are essential in troubleshooting the information gaps. These technical advancements have greatly expanded the candidate gene openings. The secondary metabolites being important precursors need to be studied in a much wider scale for accurate calculations of biochemical reactions, taking place inside and outside the synthesized living cell. The present review highlights the sequencing techniques to provide a foundation of opportunity generation for agricultural sustainability.

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Salinization poses a significant challenge in agriculture, exacerbated by anthropogenic global warming. Biostimulants, derived from living microorganisms or natural extracts, have emerged as valuable tools for conventional and organic agriculture. However, our understanding of the molecular mechanisms underlying the effects of biostimulants is very limited, especially in crops under real cultivation conditions. In this study, we adopted an integrative approach to investigate the effectiveness of the combined application of plant growth-promoting bacterium (Bacillus megaterium strain BM08) and a non-microbial biostimulant under control conditions (normal watering) and salt stress. After confirming the yield increase under both conditions, we investigated the molecular mechanisms underlying the observed effect by measuring a number of physiological parameters (i.e., lipid peroxidation, antioxidants, chlorophylls, total phenolics and phytohormone content), as well as RNA sequencing and primary metabolite analyses. Our findings reveal that the combined effect of the microbial and non-microbial biostimulants led to a decrease in the antioxidant response and an up-regulation of genes involved in cytokinin biosynthesis under salt stress conditions. This, in turn, resulted in a higher concentration of the bioactive cytokinin, isopentenyladenosine, in roots and leaves and an increase in γ-aminobutyric acid, a non-proteic amino acid related to abiotic stress responses. In addition, we observed a decrease in malic acid, along with an abscisic acid (ABA)-independent up-regulation of SR-kinases, a family of protein kinases associated with abiotic stress responses. Furthermore, we observed that the single application of the non-microbial biostimulant triggers an ABA-dependent response under salt stress; however, when combined with the microbial biostimulant, it potentiated the mechanisms triggered by the BM08 bacterial strain. This comprehensive investigation shows that the combination of two biostimulants is able to elicit a cytokinin-dependent response that may explain the observed yield increase under salt stress conditions.

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Smoke-water (SW) and Karrikinolide1 (KAR1) release dormancy and improve seed germination in many plant species. Therefore, we tested SW (1:2500 v/v) and KAR1 (10-7 M) to break the morphological dormancy of celery cultivar (Apium graveolens L.). In the first trial, seeds were subjected to a 21-day incubation period at 20 °C with SW and KAR1 applied as single treatments. KAR1 showed significantly improved germination (30.7%) as compared to SW (17.2%) and a water control (14.7%). In seed soaking experiments, SW, KAR1, and gibberellic acid (GA3) treatments showed higher germination percentages than the water control after 3 and 6 h of soaking. However, prolonged soaking (12 h) reduced germination percentages for all treatments, indicating a detrimental effect. Analysis of KAR1 content dynamics in 7-day- and 21-day-old celery seeds indicated its prolonged effects on germination and dormancy alleviation. Phytohormones, including auxins in 7-day-old and cytokinins in 7-day- and 21-day-old celery seedlings, along with their precursors and metabolites, were analyzed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) after treatment with KAR1 and SW. The analysis of auxin levels in 7-day-old seeds revealed a negative correlation between seed germination and auxin (indole-3-acetic acid, IAA) content. Notably, it was found that KAR1-treated seeds significantly reduced IAA levels in all treatments. SW and KAR1 did not significantly affect cytokinin levels during celery germination except for N6-Isopentenyladenine. Hence, further research is needed to understand their precise role in celery seed germination. This work will improve our understanding of the role of bioactive compounds from plant-derived smoke and how they regulate hormonal responses and improve germination efficiency in celery.

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In recent years, inorganic nanoparticles, including calcium hydroxide nanoparticles [Ca Ca(OH)2 NPs], have attracted significant interest for their ability to impact plant photosynthesis and boost agricultural productivity. In this study, the effects of 15 and 30 mg L-1 oleylamine-coated calcium hydroxide nanoparticles [Ca(OH)2@OAm NPs] on photosystem II (PSII) photochemistry were investigated on tomato plants at their growth irradiance (GI) (580 µmol photons m-2 s-1) and at high irradiance (HI) (1000 µmol photons m-2 s-1). Ca(OH)2@OAm NPs synthesized via a microwave-assisted method revealed a crystallite size of 25 nm with 34% w/w of oleylamine coater, a hydrodynamic size of 145 nm, and a ζ-potential of 4 mV. Compared with the control plants (sprayed with distilled water), PSII efficiency in tomato plants sprayed with Ca(OH)2@OAm NPs declined as soon as 90 min after the spray, accompanied by a higher excess excitation energy at PSII. Nevertheless, after 72 h, the effective quantum yield of PSII electron transport (ΦPSII) in tomato plants sprayed with Ca(OH)2@OAm NPs enhanced due to both an increase in the fraction of open PSII reaction centers (qp) and to the enhancement in the excitation capture efficiency (Fv'/Fm') of these centers. However, the decrease at the same time in non-photochemical quenching (NPQ) resulted in an increased generation of reactive oxygen species (ROS). It can be concluded that Ca(OH)2@OAm NPs, by effectively regulating the non-photochemical quenching (NPQ) mechanism, enhanced the electron transport rate (ETR) and decreased the excess excitation energy in tomato leaves. The delay in the enhancement of PSII photochemistry by the calcium hydroxide NPs was less at the GI than at the HI. The enhancement of PSII function by calcium hydroxide NPs is suggested to be triggered by the NPQ mechanism that intensifies ROS generation, which is considered to be beneficial. Calcium hydroxide nanoparticles, in less than 72 h, activated a ROS regulatory network of light energy partitioning signaling that enhanced PSII function. Therefore, synthesized Ca(OH)2@OAm NPs could potentially be used as photosynthetic biostimulants to enhance crop yields, pending further testing on other plant species.

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Integrating agricultural, chemical, and technological knowledge is crucial for developing bio-nanotechnologies to improve agricultural production. This study explores the innovative use of biopolymeric coatings, based on sodium alginate and sodium alginate + Laponite® (nanoclay), containing biostimulants (tryptophol and thymol) or not, on garlic cloves. These coatings were analyzed using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR-ATR), and scanning electron microscopy (SEM). Greenhouse bioassays showed improvements in garlic shoot plant biomass with both treatments: sodium alginate biopolymer and sodium alginate biopolymer plus Laponite®. In the field experiment, garlic plants treated with sodium alginate, in combination with conventional pesticide treatments, resulted in better quality garlic bulbs, where larger garlics were harvested in this treatment, reducing commercial losses. In tropical garlic crops, obtaining plants with greater initial vigor is essential. Our results highlight the potential of these bio-nanotechnological strategies to enhance garlic propagation, ensuring environmental protection and food security.

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Salinity poses a significant challenge to tomato plant development and metabolism. This study explores the use of biostimulants as eco-friendly strategies to enhance tomato plant tolerance to salinity. Conducted in a greenhouse, the research focuses on the Solanum lycopersicum L. behavior under saline conditions. Tomato seeds were treated with arbuscular mycorrhizal fungi (AMF), compost, and their combination under both non-saline and saline conditions (0 and 150 mM NaCl). Plant height, number of flowers and fruits, shoot fresh weight, and root dry weight were negatively impacted by salt stress. The supplementation with compost affected the colonization of AMF, but the application of stress had no effect on this trait. However, the use of compost and AMF separately or in combination showed positive effects on the measured parameters. At the physiological level, compost played a beneficial role in increasing photosynthetic efficiency, whether or not plants were subjected to salinity. In addition, the application of these biostimulants led to an increase in nitrogen content in the plants, irrespective of the stress conditions. AMF and compost, applied alone or in combination, showed positive effects on photosynthetic pigment concentrations and protein content. Under salt stress, characterized by an increase in lipid peroxidation and H2O2 content, the application of these biostimulants succeeded in reducing both these parameters in affected plants through exhibiting an increase in antioxidant enzyme activity. In conclusion, incorporating compost, AMF, or their combined application emerges as a promising approach to alleviate the detrimental impacts of salt stress on both plant performances. These findings indicate optimistic possibilities for advancing sustainable and resilient agricultural practices.

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Bio-stimulants, such as selenium nanoparticles and melatonin, regulate melon growth. However, the effects of individual and combined applications of selenium nanoparticles and melatonin on the growth of melon seedlings have not been reported. Here, two melon cultivars were sprayed with selenium nanoparticles, melatonin, and a combined treatment, and physiological and biochemical properties were analyzed. The independent applications of selenium nanoparticles, melatonin, and their combination had no significant effects on the plant heights and stem diameters of Jiashi and Huangmengcui melons. Compared with the controls, both selenium nanoparticle and melatonin treatments increased soluble sugars (6-63%) and sucrose (11-88%) levels, as well as the activity of sucrose phosphate synthase (171-237%) in melon leaves. The phenylalanine ammonia lyase (29-95%), trans cinnamate 4-hydroxylase (32-100%), and 4-coumaric acid CoA ligase (26-113%), as well as mRNA levels, also increased in the phenylpropanoid metabolism pathway. Combining the selenium nanoparticles and melatonin was more effective than either of the single treatments. In addition, the levels of superoxide dismutase (43-130%), catalase (14-43%), ascorbate peroxidase (44-79%), peroxidase (25-149%), and mRNA in melon leaves treated with combined selenium nanoparticles and melatonin were higher than in controls. The results contribute to our understanding of selenium nanoparticles and melatonin as bio-stimulants that improve the melon seedlings' growth by regulating carbohydrate, polyamine, and antioxidant capacities.

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Bacterial exopolysaccharides (EPS) are extracellular polymer-based substances recently defined as potential plant biostimulants, as they can increase nutrient uptake, water retention, and resistance to abiotic stress. As sugar-based substances, the bacteria producing them need to grow in a sugar-rich substrate. Hence, some agri-food by-products could be used as suitable carbon sources for EPS production as a cost-effective and more sustainable alternative to conventional substrates. Thus, this study aimed to produce EPS from specific bacterial strains through solid-state fermentation (SSF) using agri-food waste as a low-cost substrate. Six residues and five bacterial strains were tested in a lab-scale SSF system. From the assessed substrate-strain combinations, Burkholderia cepacia with ginger juice waste (GJW) resulted in the most promising considering several process parameters (EPS production, cumulative oxygen consumption, biomass growth, reducing sugars consumption). Also, dynamic monitoring of the system allowed for establishing 5 days as a suitable fermentation time. Then, using response surface methodology (Box-Behnken design), the process was optimised based on airflow rate (AF), inoculum size (IS), and micronutrient concentration (MN). In this stage, the best conditions found were at 0.049 (± 0.014) L h-1 per gram of dry matter (DM) for AF, 8.4 (± 0.9) E + 09 CFU g-1 DM for IS, and 0.07 (± 0.01) mL g-1 DM for MN, reaching up to 71.1 (± 3.2) mg crude EPS g-1 DM. Results show the potential of this approach to provide a new perspective on the value chain for the agri-food industry by introducing it to a circular economy framework.

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An experiment was performed to evaluate the effect of Trichoderma harzianum MVT801 combined with different ratios of nutrient solution (NR) (25%, 50%, and 100%) on the growth and physiological traits of Lactuca sativa "Lollo Rosso" plants cultivated in floating systems. Inoculation of lettuce plants with T. harzianum MVT801 (T1) in a floating system improves biometric properties, photosynthetic parameters, and nutrient uptake compared with uninoculated treatment (T0). The results clearly showed that in T1, despite a 50% reduction in the ratio of nutrient solution, no significant difference was observed in the growth and photosynthesis characteristics and nutrient uptake in L. sativa "Lollo Rosso" leaves compared with a complete nutrient solution treatment (100%), which is one of the notable results of this study. In this regard, the highest yield was observed in T1NR50 (inoculated with fungi and 50% of the nutrient solution) and T1NR100 (inoculated with fungi and complete nutrient solution) treatments. Also, the highest concentrations of phosphorus and potassium in "Lollo Rosso" leaves were observed in T1NR50 and T1NR100 treatments. Accordingly, the use of T. harzianum in floating lettuce cultivation could be recommended to increase crop productivity and reduce the use of chemical fertilizers.

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Several Acacia species are aggressive invaders outside their native range, often occupying extensive areas. Traditional management approaches have proven to be ineffective and economically unfeasible, especially when dealing with large infestations. Here, we explain a different approach to complement traditional management by using the waste from Acacia management activities. This approach can provide stakeholders with tools to potentially reduce management costs and encourage proactive management actions. It also prioritizes potential applications of Acacia waste biomass for agriculture and forestry as a way of sequestering the carbon released during control actions. We advocate the use of compost/vermicompost, green manure and charcoal produced from Acacia waste, as several studies have shown their effectiveness in improving soil fertility and supporting crop growth. The use of waste and derivatives as bioherbicides or biostimulants is pending validation under field conditions. Although invasive Acacia spp. are banned from commercialization and cultivation, the use of their waste remains permissible. In this respect, we recommend the collection of Acacia waste during the vegetative stage and its subsequent use after being dried or when dead, to prevent further propagation. Moreover, it is crucial to establish a legal framework to mitigate potential risks associated with the handling and disposal of Acacia waste.

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Biostimulants and other plant growth promoters can provide an effective solution to the challenge of urbanisation and climate change. Viburnum opulus 'Roseum' is a globally popular deciduous shrub species that can be made more resistant to urban influences by using natural growth-promoting substances. In our study, we investigated the effects of growth promoters Kelpak®, Bistep and Yeald Plus on the species, both histologically and physiologically (proline stress hormone measurement). Our measurements were complemented using the analysis of rhizosphere alkaline phosphatase, ß-glucosidase and ß-glucosaminidase enzymes, to obtain a more complete picture of the combined effect of biostimulants and species. We found that the Bistep biostimulant had an outstanding effect on the leaf tissue culture results of the variety. The transpiration and evapotranspiration findings also confirmed the efficacy of biostimulants. In the case of POD activity and rhizosphere enzyme measurements, Bistep and Yeald Plus obtained statistically higher values than the control group. Kelpak produced better results than the control group in several measurements (alkaline phosphatase levels; evapotranspiration results), but in other cases it resulted in lower values than the control treatment. The use of Bistep and Yeald Plus can greatly assist growers in the cultivation of V. opulus 'Roseum' in an urban environment.

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Plant biostimulants have received attention as sustainable alternatives to chemical fertilizers. Extracellular polymeric substances (EPSs), among the compounds secreted by plant growth-promoting rhizobacteria (PGPRs), are assumed to alleviate abiotic stress. This study aims to investigate the effect of purified EPSs on rice under abiotic stress and analyze their mechanisms. A pot experiment was conducted to elucidate the effects of inoculating EPSs purified from PGPRs that increase biofilm production in the presence of sugar on rice growth in heat-stress conditions. Since all EPSs showed improvement in SPAD after the stress, Enterobacter ludwigii, which was not characterized as showing higher PGP bioactivities such as phytohormone production, nitrogen fixation, and phosphorus solubilization, was selected for further analysis. RNA extracted from the embryos of germinating seeds at 24 h post-treatment with EPSs or water was used for transcriptome analysis. The RNA-seq analysis revealed 215 differentially expressed genes (DEGs) identified in rice seeds, including 139 up-regulated and 76 down-regulated genes. A gene ontology (GO) enrichment analysis showed that the enriched GO terms are mainly associated with the ROS scavenging processes, detoxification pathways, and response to oxidative stress. For example, the expression of the gene encoding OsAAO5, which is known to function in detoxifying oxidative stress, was two times increased by EPS treatment. Moreover, EPS application improved SPAD and dry weights of shoot and root by 90%, 14%, and 27%, respectively, under drought stress and increased SPAD by 59% under salt stress. It indicates that bacterial EPSs improved plant growth under abiotic stresses. Based on our results, we consider that EPSs purified from Enterobacter ludwigii can be used to develop biostimulants for rice.

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BACKGROUND: Protein hydrolysates (PHs) can enhance plant nitrogen nutrition and improve the quality of vegetables, depending on their bioactive compounds. A tomato greenhouse experiment was conducted under both optimal (14 mM) and suboptimal (2 mM) nitrogen (N-NO3) conditions. Tomatoes were treated with a new Malvaceae-derived PH (MDPH) and its molecular fractions (MDPH1, >10 kDa; MDPH2, 1-10 kDa and MDPH3, <1 kDa). RESULTS: Under optimal N conditions, the plants increased biomass and fruit yield, and showed a higher photosynthetic pigment content in leaves in comparison with suboptimal N, whereas under N-limiting conditions, an increase in dry matter, soluble solid content (SSC) and lycopene, a reduction in firmness, and changes in organic acid and phenolic compounds were observed. With 14 mM N-NO3, MDPH3 stimulated an increase in dry weight and increased yield components and lycopene in the fruit. The MDPH2 fraction also resulted in increased lycopene accumulation in fruit under 14 mM N-NO3. At a low N level, the PH fractions showed distinct effects compared with the whole MDPH and the control, with an increase in biomass for MDPH1 and MDPH2 and a higher pigment content for MDPH3. Regardless of N availability, all the fractions affected fruit quality by increasing SSC, whereas MDPH2 and MDPH3 modified organic acid content and showed a higher concentration of flavonols, lignans, and stilbenes. CONCLUSION: The molecular weight of the peptides modifies the effect of PHs on plant performance, with different behavior depending on the level of N fertilization, confirming the effectiveness of fractioning processes. © 2024 Society of Chemical Industry.

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Climate change is currently considered as one of the main concerns of the agriculture sector, as it limits crop production and quality. Furthermore, the current context of global crisis with international political instability and war conflicts over the world is pushing the agriculture sector even more to urgently boost productivity and yield and doing so in a sustainable way in the current frame of climate change. Biostimulants can be an effective tool in alleviating the negative effects of environmental stresses to which plants are exposed, such as drought, salinity, heavy metals and extreme temperatures etc. Biostimulants act through multiple mechanisms, modifying gene expression, metabolism and phytohormone production, promoting the accumulation of compatible solutes and antioxidants and mitigating oxidative stress. However, it is important to keep in mind that the use and effect of biostimulants has limitations and must be accompanied by other techniques to ensure crop yield and quality in the current frame of climate change, such as proper crop management and the use of other sustainable resources. Here, we will not only highlight the potential use of biostimulants to face future agricultural challenges, but also take a critical look at their limitations, underlining the importance of a broad vision of sustainable agriculture in the context of climate change.

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