Salt-tolerant plant growth-promoting bacteria as a versatile tool for combating salt stress in crop plants.

Soil salinization poses a great threat to global agricultural ecosystems, and finding ways to improve the soils affected by salt and maintain soil health and sustainable productivity has become a major challenge. Various physical, chemical and biological approaches are being evaluated to address this escalating environmental issue. Among them, fully utilizing salt-tolerant plant growth-promoting bacteria (PGPB) has been labeled as a potential strategy to alleviate salt stress, since they can not only adapt well to saline soil environments but also enhance soil fertility and plant development under saline conditions. In the last few years, an increasing number of salt-tolerant PGPB have been excavated from specific ecological niches, and various mechanisms mediated by such bacterial strains, including but not limited to siderophore production, nitrogen fixation, enhanced nutrient availability, and phytohormone modulation, have been intensively studied to develop microbial inoculants in agriculture. This review outlines the positive impacts and growth-promoting mechanisms of a variety of salt-tolerant PGPB and opens up new avenues to commercialize cultivable microbes and reduce the detrimental impacts of salt stress on plant growth. Furthermore, considering the practical limitations of salt-tolerant PGPB in the implementation and potential integration of advanced biological techniques in salt-tolerant PGPB to enhance their effectiveness in promoting sustainable agriculture under salt stress are also accentuated.

Do Lignin Nanoparticles Pave the Way for a Sustainable Nanocircular Economy? Biostimulant Effect of Nanoscaled Lignin in Tomato Plants.

Agriculture has a significant environmental impact and is simultaneously called to major challenges, such as responding to the need to develop more sustainable cropping systems with higher productivity. In this context, the present study aimed to obtain lignin nanoparticles (LNs) from pomace, a waste product of the olive oil chain, to be used as a nanobiostimulant in tomato plants. The biostimulant effect of this biopolymer is known, but its reduction to nanometer size can emphasize this property. Tomato plants were subjected to different LN dosages (25, 50, and 100 mg L-1) by foliar application, and inductive effects on photosynthetic machinery, aerial and root biomass production, and root morphology were observed. The treated plants showed increased efficiency in catching and using light, while they reduced the fraction dissipated as heat or potentially toxic to cells for the possibility of creating reactive oxygen species (ROS). Finally, this benefit was matched by increased pigment content and a stimulatory action on the content of nitrogen (NBI) and antioxidant substances such as flavonoids. In conclusion, the present study broadens the horizon of substances with biostimulant action by demonstrating the validity and efficacy of nanobiostimulants obtained from biological residues from the olive oil production chain.

Rosmarinus officinalis L. essential oil enhances salt stress tolerance of durum wheat seedlings through ROS detoxification and stimulation of antioxidant defense.

Salt-induced stress poses a significant barrier to agricultural productivity by impeding crop growth. Presently, environmentalists are dedicated to safeguarding food security by enhancing agricultural yields in challenging environments. Biostimulants play a crucial role in mitigating abiotic stresses in crop production, and among these, plant essential oils (EOs) stand out as organic substances with diverse biological effects on living organisms. Among the natural promoters of plant growth, Rosmarinus officinalis L. essential oil (RoEO) has gained considerable attention. Although the manifold effects of essential oils (EOs) on plant growth have been extensively demonstrated, their impact on salt stress tolerance in durum wheat seedlings remains unexplored. This investigation was undertaken to evaluate the biostimulatory capabilities of RoEO on the durum wheat cultivar "Mahmoudi." The effects of three RoEO concentrations (1, 2.5, and 5 ppm) on seed germination, growth establishment, and the induction of salt resistance under salinity conditions (150 mM NaCl) were tested. At 5 ppm, RoEO enhanced seedlings' tolerance to salinity by improving growth and reducing membrane deterioration and oxidative stress-induced damage. The expression profile analyses of seven stress-related genes (TdNHX1, TdSOS1, TdSOD, TdCAT, TdGA20-ox1, TdNRT2.1, and TdGS) using RT-qPCR showed enhancement of several important genes in durum wheat seedlings treated with 5 ppm RoEO, even under control conditions, which may be related to salt stress tolerance. The results indicate that the application of RoEO suggests a possible alternative strategy to increase salt tolerance in durum wheat seedlings towards better growth quality, thus increasing ROS scavenging and activation of antioxidant defense.

Enhancing Lettuce Yield through Innovative Foliar Spray of Biopolymers Derived from Municipal Biowastes.

Municipal waste biomass could be valorized as an alternative feedstock to produce compounds beneficial for agricultural applications. The foliar spray application of biostimulants emerges as a promising and innovative technique due to its environmental safety and ability to enhance crop yields. In recent years, the exploitation of biopolymers obtained through alkaline hydrolysis of the solid anaerobic digestate from municipal biowastes has attracted researchers' interest. The aim of this study is to investigate the effects on lettuce growth of a product obtained through alkaline hydrolysis from municipal biowaste, Biopolymers (BPs), and of a derivate subjected to a further oxidation process, Biopolymers Oxidate (BPs OX). The effects of the treatments at various concentrations were evaluated by monitoring plant growth and observing the trends in the activities of the main enzymes involved in the nitrogen metabolic pathway of lettuce. Results suggest that the best treatments in terms of fresh weight were achieved by using BPs at 10 mg/L and BPs OX at 100 mg/L, increasing yield by around 28% and 34%, respectively. The innovative aspect of this work was to make easier for farmers the biopolymers application by testing a foliar spray methodology for BPs and BPs OX, which has never been tested before in any crop.

Isoptericola haloaureus sp. nov., a dimorphic actinobacterium isolated from mangrove sediments of southeast India, implicating biosaline agricultural significance through nitrogen fixation and salt tolerance genes.

Strain MP-1014T, an obligate halophilic actinobacterium, was isolated from the mangrove soil of Thandavarayancholanganpettai, Tamil Nadu, India. A polyphasic approach was utilized to explore its phylogenetic position completely. The isolate was Gram-positive, filamentous, non-motile, and coccoid in older cultures. Ideal growth conditions were seen at 30 °C and pH 7.0, with 5% NaCl (W/V), and the DNA G + C content was 73.3%. The phylogenic analysis of this strain based upon 16S rRNA gene sequence revealed 97-99.8% similarity to the recognized species of the genus Isoptericola. Strain MP-1014T exhibits the highest similarity to I. sediminis JC619T (99.7%), I. chiayiensis KCTC19740T (98.9%), and subsequently to I. halotolerans KCTC19646T (98.6%), when compared with other members within the Isoptericola genus (< 98%). ANI scores of strain MP-1014T are 86.4%, 84.2%, and 81.5% and dDDH values are 59.7%, 53.6%, and 34.8% with I. sediminis JC619T, I. chiayiensis KCTC19740T and I. halotolerans KCTC19646T respectively. The major polar lipids of the strain MP-1014T were phosphatidylinositol, phosphatidylglycerol, diphosphotidylglycerol, two unknown phospholipids, and glycolipids. The predominant respiratory menaquinones were MK9 (H4) and MK9 (H2). The major fatty acids were anteiso-C15:0, anteiso-C17:0, iso-C14:0, C15:0, and C16:0. Also, initial genome analysis of the organism suggests it as a biostimulant for enhancing agriculture in saline environments. Based on phenotypic and genetic distinctiveness, the strain MP-1014 T represents the novel species of the genus Isoptericola assigned Isoptericola haloaureus sp. nov., is addressed by the strain MP-1014 T, given its phenotypic, phylogenetic, and hereditary uniqueness. The type strain is MP-1014T [(NCBI = OP672482.1 = GCA_036689775.1) ATCC = BAA 2646T; DSMZ = 29325T; MTCC = 13246T].

Dataset on effect of biostimulants on growth and rooting of kiwifruit (Actinidia deliciosa) cuttings: from morphological and gene regulation approach.

Actinidia chinensis Planch. and A. deliciosa (A. Chev.) C.F. Liang et A.R. Ferguson are the botanical names for the two main closely related kiwifruit species that are cultivated worldwide [1]. According to the Food and Agriculture Organisation (FAO) of the United Nation, kiwifruit is produced on 268,788 hectares of land worldwide, yielding 4,348,011 metric tonnes of fruit per year. China is the world's top producer, followed by Italy, New Zealand, Chile, and Greece, with a cumulative valuation of 2,907,580 thousand US dollars for export (http://www.fao.org/faostat/en/#data/QC). Several research using nutrient medium and other inorganic treatments on softwood cuttings for micro-propagation techniques have shown promising outcomes [2,3]. Several agricultural and horticultural crops have demonstrated significantly improved crop growth, quality, and reproduction when treated with seaweed extracts [4]. It is possible to utilise seaweed extracts to encourage cuttings from perennial fruit species, such as kiwifruit (Actinidia deliciosa), to root and flourish. Absence of a growth regulator permitted by organic methods is one of the main obstacles in kiwifruit production. Hardwood cuttings are the most popular technique of clonally reproducing kiwifruit, and the cuttings' ability to root depends on the application of synthetic auxins, which is not allowed in organic agriculture. Therefore, alternative biostimulants have been used to promote the rooting of kiwifruit cuttings in this study. For six hours, the cuttings in this investigation were submerged in base dipping solutions containing 1, 5, 10, and 50 % of G Sap (Gracilaria edulis), K Sap (Kappaphycus alvarezii), AN (Ascophyllum nodosum), EM (Ecklonia maxima), HA (Humic acid), and control (water). After that, for a period of six months, the treatments of G Sap, K Sap, AN, EM, HA, and control were applied (at the rate of 50 ml of solutions) to the potted cuttings at intervals of fifteen days. The dataset provided the data of the rooting percent in all the kiwifruit cultivars, namely 'Monty', 'Abott', 'Hayward', 'Allison' and 'Bruno' (P ≤ 0.01), shoot and root growth parameters including leaf number per cutting, number of roots per cutting, number of branches, plant height, shoot diameter, root length, root diameter and root weight with the application of seaweed extracts. Also data of pigments (chlorophyll a, chlorophyll b and total carotenoids), metabolites (total carbohydrates and soluble phenols) and electrolyte leakage were collected after the treatments. Data of four root promoting candidate genes (GH3-3, LBD16, LBD29 and LRP1) were also described which indicated the influence of the biostimulants on the cuttings. The application of seaweed extracts resulted in a positive increase in all shoot and root growth parameters, including the number of leaves per cutting, the number of roots per cutting, the number of branches, plant height, shoot diameter, root length, root diameter, and root weight (P ≤ 0.05). In comparison to the control cuttings, the seaweed extract-treated cuttings showed significantly greater levels of pigments (such as chlorophyll a, chlorophyll b, and total carotenoids), metabolites (such as total carbohydrates and soluble phenols), and reduced electrolyte leakage. Various treatments (1, 5 and 10% solutions of G Sap, K Sap, AN, EM, HA and control) gave positive impact on nutrient parameters of kiwifruit cultivar 'Hayward'. Moreover, the relative positive expressions of root inducing genes (GH3-3, LBD16, LBD29 and LRP1) was observed in leaves and roots of cultivar 'Hayward' by qRT-PCR after treatment with G Sap, K Sap, AN, EM, HA @ 10 % and control. Thus, it can be said that seaweed extract and humic acid are good substitutes for synthetic hormones in encouraging kiwifruit cuttings to root and flourish.

The cepacian-like exopolysaccharide of Paraburkholderia ultramafica STM10279T enhances growth and metal adaptation of Tetraria comosa on New Caledonian ultramafic soil.

Paraburkholderia ultramafica STM10279T is a metal-tolerant rhizobacterium that promotes plant growth. It was isolated from the roots of Tetraria arundinaceae, a pioneer endemic tropical herb growing on ultramafic soils in New Caledonia. We have recently shown that the main mechanism of metal tolerance of P. ultramafica is related to the production of an acidic exopolysaccharide (EPS). To explore the potential role of this EPS in the plant's environmental adaptation, we first elucidated its structure by employing a combination of chromatography and mass spectrometry techniques. These analyses revealed that the EPS is highly branched and composed of galactosyl (35.8%), glucosyl (33.2%), rhamnosyl (19.5%), mannosyl (7.2%), and glucuronosyl residues (4.4%), similar to the EPS of the Burkholderia cepacia complex known as cepacian. We subsequently conducted greenhouse experiments on Tetraria comosa plantlets inoculated with P. ultramafica or a solution of its EPS during transplanting onto ultramafic substrate. The data showed that the dry weight of T. comosa shoots was 2.5 times higher in the plants treated with the EPS compared to the unexposed plants. In addition, inductively coupled plasma-optical emission spectrometry (ICP-OES) analysis revealed that exposure to the EPS significantly increased Ca, Mg, K, and P uptake as well as K content in roots. In vitro experiments using the Pikovskaya method showed that the EPS was able to solubilize phosphorus. Consistent with the retention of metals in roots and a reduction in shoots, our data revealed a significant decrease in metal translocation factors (TFs) in the plants inoculated with the EPS. These results suggest a beneficial effect of the rhizobacterial EPS on plant growth and abiotic stress mitigation. In addition, the data suggest that the reduced levels of trace metals in plants exposed to P. ultramafica STM10279T are due to metal chelation by the EPS. Further investigations are needed to firmly demonstrate whether this EPS could be used as a biostimulant for plant growth and adaptation to ultramafic soils.

A new biostimulant derived from soybean by-products enhances plant tolerance to abiotic stress triggered by ozone.

BACKGROUND: Tropospheric ozone is an air pollutant that causes negative effects on vegetation, leading to significant losses in crop productivity. It is generated by chemical reactions in the presence of sunlight between primary pollutants resulting from human activity, such as nitrogen oxides and volatile organic compounds. Due to the constantly increasing emission of ozone precursors, together with the influence of a warming climate on ozone levels, crop losses may be aggravated in the future. Therefore, the search for solutions to mitigate these losses becomes a priority. Ozone-induced abiotic stress is mainly due to reactive oxygen species generated by the spontaneous decomposition of ozone once it reaches the apoplast. In this regard, compounds with antioxidant activity offer a viable option to alleviate ozone-induced damage. Using enzymatic technology, we have developed a process that enables the production of an extract with biostimulant properties from okara, an industrial soybean byproduct. The biostimulant, named as OEE (Okara Enzymatic Extract), is water-soluble and is enriched in bioactive compounds present in okara, such as isoflavones. Additionally, it contains a significant fraction of protein hydrolysates contributing to its functional effect. Given its antioxidant capacity, we aimed to investigate whether OEE could alleviate ozone-induced damage in plants. For that, pepper plants (Capsicum annuum) exposed to ozone were treated with a foliar application of OEE. RESULTS: OEE mitigated ozone-induced damage, as evidenced by the net photosynthetic rate, electron transport rate, effective quantum yield of PSII, and delayed fluorescence. This protection was confirmed by the level of expression of genes associated with photosystem II. The beneficial effect was primarily due to its antioxidant activity, as evidenced by the lipid peroxidation rate measured through malondialdehyde content. Additionally, OEE triggered a mild oxidative response, indicated by increased activities of antioxidant enzymes in leaves (catalase, superoxide dismutase, and guaiacol peroxidase) and the oxidative stress index, providing further protection against ozone-induced stress. CONCLUSIONS: The present results support that OEE protects plants from ozone exposure. Taking into consideration that the promotion of plant resistance against abiotic damage is an important goal of biostimulants, we assume that its use as a new biostimulant could be considered.

Biostimulant Response of Foliar Application of Rare Earth Elements on Physiology, Growth, and Yield of Rice.

Rare earth elements (REEs) have been intentionally used in Chinese agriculture since the 1980s to improve crop yields. Around the world, REEs are also involuntarily applied to soils through phosphate fertilizers. These elements are known to alleviate damage in plants under abiotic stresses, yet there is no information on how these elements act in the physiology of plants. The REE mode of action falls within the scope of the hormesis effect, with low-dose stimulation and high-dose adverse reactions. This study aimed to verify how REEs affect rice plants' physiology to test the threshold dose at which REEs could act as biostimulants in these plants. In experiment 1, 0.411 kg ha-1 (foliar application) of a mixture of REE (containing 41.38% Ce, 23.95% La, 13.58% Pr, and 4.32% Nd) was applied, as well as two products containing 41.38% Ce and 23.95% La separately. The characteristics of chlorophyll a fluorescence, gas exchanges, SPAD index, and biomass (pot conditions) were evaluated. For experiment 2, increasing rates of the REE mix (0, 0.1, 0.225, 0.5, and 1 kg ha-1) (field conditions) were used to study their effect on rice grain yield and nutrient concentration of rice leaves. Adding REEs to plants increased biomass production (23% with Ce, 31% with La, and 63% with REE Mix application) due to improved photosynthetic rate (8% with Ce, 15% with La, and 27% with REE mix), favored by the higher electronic flow (photosynthetic electron transport chain) (increase of 17%) and by the higher Fv/Fm (increase of 14%) and quantum yield of photosystem II (increase of 20% with Ce and La, and 29% with REE Mix), as well as by increased stomatal conductance (increase of 36%) and SPAD index (increase of 10% with Ce, 12% with La, and 15% with REE mix). Moreover, adding REEs potentiated the photosynthetic process by increasing rice leaves' N, Mg, K, and Mn concentrations (24-46%). The dose for the higher rice grain yield (an increase of 113%) was estimated for the REE mix at 0.72 kg ha-1.

Yield and Fruit Characteristics of Tomato Crops Grown with Mineral Macronutrients: Impact of Organo-Mineral Fertilizers through Foliar or Soil Applications.

The utilization of plant biostimulants has gained importance as a strategy by which to improve plant productivity and soil health. Two independent trials were conducted across two seasons (2021 and 2023) to evaluate the effects of foliar or soil applications of various commercial organo-mineral fertilizers (Futuroot®, Radicon® Amifort®) with biostimulant action that is exerted on yield and fruit characteristics of processing tomato crops (cv Taylor F1) that have been exposed to mineral macronutrients. These treatments were administered three times during the season: at the transplanting, pre-flowering and berry development stages. In the first trial, conducted in two fields characterized respectively by low and high fertility, foliar applications of Radicon®, which is based on humic acid and amino acids, increased the leaf greenness index SPAD compared with the control. Furthermore, the leaf green colour intensity (SPAD index), measured during the reproductive phases of the tomato exhibited a positive correlation (R2 = 0.726) with the marketable yield obtained. This increase in marketable yield was significant in the biostimulant treatment compared with the control in both soils, especially in the soil characterized by lower fertility (16.1%), when compared with the more fertile soil (6.8%). In the second trial, conducted in the low-fertility field mentioned above, soil applications of all biostimulants (Futuroot®, Radicon® and the combinations [Radicon® + Amifort-Plus®]) significantly increased the marketable yield by 27.8%, 13.5% and 27.7%, respectively, compared with the control. The most significant beneficial effects of both Futuroot® and [Radicon® + Amifort®] could be attributed to the combination of humic acids and auxins, cytokinins or microelements (Zn, Mn, MgO) present in the formulation of these products. Furthermore, the increase in marketable yield obtained when Radicon® was applied to leaves was higher (16.1%) than that observed with soil application (13.5%). In both trials, no relevant effects of biostimulant products were observed on most of the physicochemical characteristics of tomato fruits. In conclusion, the biostimulants based on humic acid and amino acids combined with chemical fertilizers tested in the present study and applied by fertigation were more effective in improving tomato yield, and therefore they can be recommended for efficient agricultural production.

Chitosan derivatives act as a bio-stimulants in plants: A review.

Chitosan has been considered an eco-friendly biopolymer. Chitosan is a natural polycationic linear polysaccharide composed of D-glucosamine and N-acetyl-D-glucosamine linked by ß-1,4-glycosidic bonds. Chitosan has been used as an eco-friendly biopolymer for so many agricultural applications. Unfortunately, the relatively poor solubility and poor antimicrobial properties limit its widespread applications in agriculture sciences. Hence, chitosan derivatives are produced via various chemical approaches such as cross-linking, carboxylation, ionic binding, and so on. As an alternative to chemical fertilizers, chitosan derivatives, chitosan conjugates, nanostructures, semisynthetic derivatives, oligo mixes, chitosan nanoparticles, and chitosan nano-carriers are synthesized for various agricultural applications. Its several chemical and physical properties such as biocompatibility, biodegradability, permeability, cost-effectiveness, low toxicity, and environmental friendliness make it useful for many agricultural applications. Hence, popularizing its use as an elicitor molecule for different host-pathogen interaction studies. Thus, the versatile and plethora of chitosan derivatives are gaining momentum in agricultural sciences. Bio-stimulant properties and multifunctional benefits are associated with further prospective research. Therefore, in the present review, we decipher the potential pros and cons of chitosan derivatives in plants.

Cytokinin oxidase/dehydrogenase inhibitors: progress towards agricultural practice.

Cytokinin oxidase/dehydrogenase (CKX) inhibitors reduce the degradation of cytokinins in plants and thereby may improve the efficiency of agriculture and plant tissue culture-based practices. Here, we report a synthesis and structure-activity relationship study of novel urea derivatives concerning their CKX inhibitory activity. The best compounds showed sub-nanomolar IC50 values with maize ZmCKX1, the lowest value yet documented. Other CKX isoforms of maize (Zea mays) and Arabidopsis were also inhibited very effectively. The binding mode of four compounds was characterized based on high-resolution crystal complex structures. Using the soil nematode Caenorhabditis elegans, and human skin fibroblasts, key CKX inhibitors with low toxicity were identified. These compounds enhanced the shoot regeneration of Lobelia, Drosera, and Plectranthus, as well as the growth of Arabidopsis and Brassica napus. At the same time, a key compound (namely 82), activated a cytokinin primary response gene ARR5:GUS and cytokinin sensor TCSv2:GUS, without activating the Arabidopsis cytokinin receptors AHK3 and AHK4. This strongly implies that the effect of compound 82 is due to the upregulation of cytokinin signalling. Overall, this work presents highly effective and easily prepared CKX inhibitors with a low risk of environmental toxicity for further investigation of their potential in agriculture and biotechnology.

Enhancing high-density microalgae cultivation via exogenous supplementation of biostimulant derived from onion peel waste for sustainable biodiesel production.

Microalgae demonstrate significant potential as a source of liquid-based biofuels. However, increasing biomass productivity in existing cultivation systems is a critical prerequisite for their successful integration into large-scale operations. Thus, the current work aimed to accelerate the growth of C. vulgaris via exogenous supplementation of biostimulant derived from onion peel waste. Under the optimal growth conditions, which entailed a biostimulant dosage of 37.5% v/v, a pH of 3, an air flow rate of 0.4 L/min, and a 2% v/v inoculum harvested during the mid-log phase, yielded a maximum biomass concentration of 1.865 g/L. Under the arbitrarily optimized parameters, a comparable growth pattern was evident in the upscaled cultivation of C. vulgaris, underscoring the potential commercial viability of the biostimulant. The biostimulant, characterized through gas chromatography-mass spectrometry (GC-MS) analysis, revealed a composition rich in polyphenolic and organo-sulphur compounds, notably including allyl trisulfide (28.13%), methyl allyl trisulfide (23.04%), and allyl disulfide (20.78%), showcasing potent antioxidant properties. Additionally, microalgae treated with the biostimulant consistently retained their lipid content at 18.44% without any significant reduction. Furthermore, a significant rise in saturated fatty acid (SFA) content was observed, with C16:0 and C18:1 dominating both bench-scale (44.08% and 14.01%) and upscaled (51.12% and 13.07%) microalgae cultures, in contrast to the control group where C18:2 was prevalent. Consequently, SFA contents reached 54.35% and 65.43% in bench-scale and upscaled samples respectively, compared to 33.73% in the control culture. These compositional characteristics align well with the requirements for producing high-quality crude biodiesel.

Integrated multi-omic approach reveals the effect of a Graminaceae-derived biostimulant and its lighter fraction on salt-stressed lettuce plants.

Plant biostimulants are widely applied in agriculture for their ability to improve plant fitness. In the present work, the impact of Graminaceae-derived protein hydrolysate (P) and its lighter molecular fraction F3 (< 1 kDa) on lettuce plants, subjected to either no salt or high salt conditions, was investigated through the combination of metabolomics and transcriptomics. The results showed that both treatments significantly modulated the transcriptome and metabolome of plants under salinity stress, highlighting an induction of the hormonal response. Nevertheless, P and F3 also displayed several peculiarities. F3 specifically modulated the response to ethylene and MAPK signaling pathway, whereas P treatment induced a down-accumulation of secondary metabolites, albeit genes controlling the biosynthesis of osmoprotectants and antioxidants were up-regulated. Moreover, according with the auxin response modulation, P promoted cell wall biogenesis and plasticity in salt-stressed plants. Notably, our data also outlined an epigenetic control of gene expression induced by P treatment. Contrarily, experimental data are just partially in agreement when not stressed plants, treated with P or F3, were considered. Indeed, the reduced accumulation of secondary metabolites and the analyses of hormone pathways modulation would suggest a preferential allocation of resources towards growth, that is not coherent with the down-regulation of the photosynthetic machinery, the CO2 assimilation rate and leaves biomass. In conclusion, our data demonstrate that, although they might activate different mechanisms, both the P and F3 can result in similar benefits, as far as the accumulation of protective osmolytes and the enhanced tolerance to oxidative stress are concerned. Notably, the F3 fraction exhibits slightly greater growth promotion effects under high salt conditions. Most importantly, this research further corroborates that biostimulants' mode of action is dependent on plants' physiological status and their composition, underscoring the importance of investigating the bioactivity of the different molecular components to design tailored applications for the agricultural practice.

Transformation of agricultural wastes into functional oligosaccharides using enzymes and emerging technologies.

INTRODUCTION: Pectin-oligosaccharides (POS) serve diverse purposes as a food ingredient, antimicrobial and biostimulant in plants, and their functionality is linked to the degree of esterification. Grape and broccoli wastes emerge as environmentally friendly alternatives to obtaining pectin, serving as a sustainable source to producing POS. For example, microwaves have proven to be an effective and sustainable method to extract polysaccharides from plant matrices. OBJECTIVE: This work aims to use grape and broccoli wastes as alternative sources for obtaining pectin by microwave-assisted extraction and biotransformation into POS, which possess biological properties. MATERIAL AND METHODS: The extraction conditions were identified at a power of 400 W, 300 s for the extraction of pectin from grape pomace and broccoli waste. Biotransformation of pectins into POS, using commercial enzyme preparations (Viscozyme L and Pectinase). Characterisation was carried out by Fourier-transform infrared spectroscopy, thermogravimetric analysis, and scanning electron microscopy. RESULTS: Physicochemical analysis indicated grape pomace and broccoli waste pectins had galacturonic acid content of 63.81 ± 1.67 and 40.83 ± 2.85 mg 100 mg-1, low degree of esterification of 34.89% and 16.22%, respectively. Biotransformation of pectins into POS resulted in a 20% hydrolysis rate. The main enzymatic activity was polygalacturonase for the degradation of the main structure of the pectin. CONCLUSION: Production of POS from agro-industrial wastes by emerging technologies, such as the combined use of microwave-assisted extraction and enzymatic processes, represents an alternative method for the generation of bioactive compounds with distinctive properties suitable for different applications of interest.

Investigation into the role of carboxylic acid and phenolic hydroxyl groups in the plant biostimulant activity of a humic acid purified from an oxidized sub-bituminous coal.

Introduction: Humic substances (HS) are increasingly being applied as crop plant biostimulants because they have been shown to increase plant productivity, especially under environmentally stressful conditions. There has been intense interest in elucidating the HS molecular structures responsible for eliciting the plant biostimulant response (PBR). The polar and weakly acidic carboxylic (COOH) and phenolic hydroxyl (ArOH) functional groups play major roles in the acid nature, pH dependent solubilities, conformation, and metal- and salt-binding capabilities of HS. Reports on the role played by these groups in the PBR of HS found growth parameters being both positively and negatively correlated with COOH and ArOH functionalities. Materials and methods: To investigate the role of COOH and ArOH in HS biostimulant activity we used a humic acid (HA), purified from an oxidized sub bituminous coal to prepare HAs with COOH groups methylated (AHA), ArOH groups acetylated (OHA), and with both COOH and ArOH groups methylated (FHA). The original HA was designated (NHA). The four HAs were subjected to elemental, 13C-NMR, FTIR, and EPR analyses and their antioxidant properties were assessed using the trolox equivalents antioxidant capacity assay (TEAC). 13C-NMR and FTIR analysis revealed significant alkylation/acetylation. To determine the effects of alkylating/acetylating these functional groups on the HA elicited PBR, the HAs were evaluated in a plant bioassay on corn (Zea mays L.) seedling under nutrient and non-nutrient stressed conditions. Treatments consisted of the four HAs applied to the soil surface at a concentration of 80 mg C L-1, in 50 ml DI H2O with the control plants receiving 50ml DI H2O. Results: The HA-treated plants, at both fertilization rates, were almost always significantly larger than their respective control plants. However, the differences produced under nutrient stress were always much greater than those produced under nutrient sufficiency, supporting previous reports that HA can reduce the effects of stress on plant growth. In addition, for the most part, the HAs with the alkylated/acetylated groups produced plants equal to or larger than plants treated with NHA. Conclusion: These results suggests that COOH and ArOH groups play a limited or no role in the HA elicited PBR. Alternatively, the HA pro-oxidant to antioxidant ratio may play a role in the magnitude of the biostimulant response.

Phenotypical and biochemical characterization of tomato plants treated with triacontanol.

Biostimulants are heterogeneous products designed to support plant development and to improve the yield and quality of crops. Here, we focused on the effects of triacontanol, a promising biostimulant found in cuticle waxes, on tomato growth and productivity. We examined various phenological traits related to vegetative growth, flowering and fruit yield, the metabolic profile of fruits, and the response of triacontanol-treated plants to salt stress. Additionally, a proteomic analysis was conducted to clarify the molecular mechanisms underlying triacontanol action. Triacontanol application induced advanced and increased blooming without affecting plant growth. Biochemical analyses of fruits showed minimal changes in nutritional properties. The treatment also increased the germination rate of seeds by altering hormone homeostasis and reduced salt stress-induced damage. Proteomics analysis of leaves revealed that triacontanol increased the abundance of proteins related to development and abiotic stress, while down-regulating proteins involved in biotic stress resistance. The proteome of the fruits was not significantly affected by triacontanol, confirming that biostimulation did not alter the nutritional properties of fruits. Overall, our findings provide evidence of the effects of triacontanol on growth, development, and stress tolerance, shedding light on its mechanism of action and providing new insights into its potential in agricultural practices.

Whole-genome sequence of the plant-associated bacterium Pseudomonas granadensis CT364 isolated in Seville, Spain.

Pseudomonas sp. CT364 was isolated from olive tree rhizosphere in Seville (Spain). We report its complete genome sequence, acquired by co-assembling Illumina and Nanopore reads. The genome comprises a circular chromosome of 6.2 Mbp and a G + C content of 60.0%. Taxonomic analyses confirmed it to be Pseudomonas granadensis.

A practical guide to the discovery of biomolecules with biostimulant activity.

The growing demand for sustainable solutions in agriculture, which are critical for crop productivity and food quality in the face of climate change and the need to reduce agrochemical usage, has brought biostimulants into the spotlight as valuable tools for regenerative agriculture. With their diverse biological activities, biostimulants can contribute to crop growth, nutrient use efficiency, and abiotic stress resilience, as well as to the restoration of soil health. Biomolecules include humic substances, protein lysates, phenolics, and carbohydrates have undergone thorough investigation because of their demonstrated biostimulant activities. Here, we review the process of the discovery and development of extract-based biostimulants, and propose a practical step-by-step pipeline that starts with initial identification of biomolecules, followed by extraction and isolation, determination of bioactivity, identification of active compound(s), elucidation of mechanisms, formulation, and assessment of effectiveness. The different steps generate a roadmap that aims to expedite the transfer of interdisciplinary knowledge from laboratory-scale studies to pilot-scale production in practical scenarios that are aligned with the prevailing regulatory frameworks.

Deficit Irrigation with Silicon Application as Strategy to Increase Yield, Photosynthesis and Water Productivity in Lettuce Crops.

In regions where water is a limited resource, lettuce production can be challenging. To address this, water management strategies like deficit irrigation are used to improve water-use efficiency in agriculture. Associating this strategy with silicon (Si) application could help maintain adequate levels of agricultural production even with limited water availability. Two lettuce crop cycles were conducted in a completely randomized design, with a factorial scheme (2 × 3), with three irrigation levels (60%, 80% and 100%) of crop evapotranspiration (ETc), and with and without Si application. To explore their combined effects, morphological, productive, physiological and nutritional parameters were evaluated in the crops. The results showed that deficit irrigation and Si application had a positive interaction: lettuce yield of the treatment with 80% ETc + Si was statistically similar to 100% ETc without Si in the first cycle, and the treatment with 60% ETc + Si was similar to 100% ETc without Si in the second cycle. Photosynthetic rate, stomatal conductance, intercellular CO2 concentration, transpiration rate and total chlorophyll content increased under water-stress conditions with Si application; in the first cycle, the treatment with 80% ETc + Si increased by 30.1%, 31.3%, 7.8%, 28.46% and 50.3% compared to the same treatment without Si, respectively. Si application in conditions of water deficit was also beneficial to obtain a cooler canopy temperature and leaves with higher relative water content. In conclusion, we found that Si applications attenuate water deficit effects and provide a strategy to ameliorate the yield and water productivity in lettuce crops, contributing to more sustainable practices in agriculture.