Effects of vegetal- versus animal-derived protein hydrolysate on sweet basil morpho-physiological and metabolic traits

Despite scientific evidence supporting the biostimulant activity of protein hydrolysates (PHs) derived from vegetal or animal sources, the morpho-physiological and biochemical mechanisms underlying the biostimulant action of PHs from plant biomass or animal by-products are still poorly explored. Accordingly, we performed a greenhouse experiment for assessing the morphological, physiological and biochemical responses of sweet basil (Ocimum basilicum L.) to three nitrogen equivalent rates (0.05, 0.15, and 0.25 g N/kg) of an animal-derived protein hydrolysate (A-PH) and a vegetal-derived protein hydrolysate (V-PH). The V-PH and A-PH applications determined a quadratic-dose response regarding the number and area of leaves and the shoot fresh and dry weight, with the best results obtained using V-PH at the N equivalent rates of 0.05 and 0.15 g N/kg. Improvement of shoot fresh weight with V-PH foliar application at the rate of 0.15 g N/kg was associated with a higher leaf CO2 assimilation and water use efficiency, with a concomitant higher uptake and translocation of K, Mg, and S in leaf tissue. The excessive accumulation of Na, Cl, and some amino acids (e.g., proline) under A-PH applications above 0.05 g N/kg induced a rapid decrease in plant photosynthetic performance, growth, and biomass production. The plants treated with A-PH at a higher dosage appeared to activate an alternative pathway involving the synthesis of alanine and GABA for storing excess ammonia, buffering cytoplasmic acidosis, and counteracting the negative effects of Na and Cl at toxic levels. The above findings demonstrated the potential benefits of protein hydrolysate application in agriculture, especially of vegetal-derived PHs, and highlighted the need to understand dose-dependent effects in order to optimize crop response

Genome Sequencing of <i>Pantoea agglomerans</i> C1 Provides Insights into Molecular and Genetic Mechanisms of Plant Growth-Promotion and Tolerance to Heavy Metals

Distinctive strains of Pantoea are used as soil inoculants for their ability to promote plant growth. Pantoea agglomerans strain C1, previously isolated from the phyllosphere of lettuce, can produce indole-3-acetic acid (IAA), solubilize phosphate, and inhibit plant pathogens, such as Erwinia amylovora. In this paper, the complete genome sequence of strain C1 is reported. In addition, experimental evidence is provided on how the strain tolerates arsenate As (V) up to 100 mM, and on how secreted metabolites like IAA and siderophores act as biostimulants in tomato cuttings. The strain has a circular chromosome and two prophages for a total genome of 4,846,925-bp, with a DNA G+C content of 55.2%. Genes related to plant growth promotion and biocontrol activity, such as those associated with IAA and spermidine synthesis, solubilization of inorganic phosphate, acquisition of ferrous iron, and production of volatile organic compounds, siderophores and GABA, were found in the genome of strain C1. Genome analysis also provided better understanding of the mechanisms underlying strain resistance to multiple toxic heavy metals and transmission of these genes by horizontal gene transfer. Findings suggested that strain C1 exhibits high biotechnological potential as plant growth-promoting bacterium in heavy metal polluted soils.6s

Protein Hydrolysates from Animal or Vegetal Sources Affect Morpho-Physiological Traits, Ornamental Quality, Mineral Composition, and Shelf-Life of Chrysanthemum in a Distinctive Manner

Protein hydrolysates (PHs) are a prominent category of plant biostimulants, mainly constituted of amino acids, oligopeptides and polypeptides, obtained by partial hydrolysis of animal or plant protein sources. Despite scientific evidence supporting the biostimulant action of PHs on vegetables, the morphological, physiological, and shelf-life performances underlying the PH action on cut flowers are still poorly explored. Accordingly, the aim of this research is to assess the effects of three commercial biostimulants, one animal PH (PH A, Hicure®) and two plant PHs (PH V1, Trainer® and PH V2, Vegamin©), on two chrysanthemum (Chrysanthemum morifolium) cultivars (Pinacolada and Radost). In both cultivars, only the plant-derived PH (V1 and V2) treatments recorded significantly higher fresh plant biomass than the control (on average +18%, in both cultivars). The foliar application of the vegetal-derived PHs but not the animal one, particularly in Pinacolada, improved the status of plants, stimulating stem elongation and the apical flower diameter. In Pinacolada, applications with PH V1 resulted in a significant increase in nitrate and P concentration in leaves and Ca content in flowers compared with the control (+43%, +27%, and +28% for nitrate, P, and Ca, respectively). In Radost, PH A and PH V2 applications caused a significant reduction in nitrate concentration in both leaves and flowers compared with the control. One week after harvest, in both cultivars, PH A applications caused flower stems to wilt faster than the control. In contrast, plants treated with PH V1 revealed significantly slower flower stem senescence compared to the control. Flower wilting during vase life was correlated to a decrease in the K-to-Na ratio in flowers due to an inability to transport K to the flowers from the leaves rather than an increase in Na in the flowers themselves

Improving Nitrogen Use Efficiency in Melon by Grafting

Identification of rootstocks capable of improving the nitrogen use efficiency (NUE) of the scion could reduce N fertilization and nitrate leaching; however, screening different graft combinations under field conditions can be costly and time-consuming. This study evaluated a rapid and economical methodology for screening of melon rootstocks for NUE. Two experiments were designed. In the first, melon plants (Cucumis melo L. cv. Proteo) either ungrafted or grafted onto four commercial rootstocks: 'Dinero' and 'Jador' (Cucumis melo L.), 'P360', and 'PS1313' (Cucurbita maxima Duchesne × Cucurbita moschata Duchesne) grown in hydroponics were compared in terms of shoot dry biomass, leaf area, root-to-shoot ratio, SPAD index, shoot N uptake, and nitrate reductase (NR) activity at the early developmental stage in response to nitrate availability (0.5, 2.5, 5, 10, or 15 mm of NO3–). The second experiment was aimed to confirm whether the use of a selected rootstock with high NUE ('P360') could improve crop performance and NUE of grafted melon plants under field conditions. In the first experiment, carried out under greenhouse conditions, melon plants grafted onto 'Dinero', 'Jador', and 'P360' rootstocks needed 5.7, 5.2, and 6.1 mm of NO3–, respectively, to reach half-maximum shoot dry weight, whereas plants grafted onto 'PS1313' rootstock and the control treatment (ungrafted plants) needed 9.1 and 13.1 mm of NO3–, respectively. Total leaf area, SPAD index, and shoot N uptake increased linearly and quadratically in response to an increase of the N concentration in the nutrient solution. At 2.5 mm of NO3–, melon plants grafted onto both C. melo and Cucurbita maxima × Cucurbita moschata rootstocks had the highest NR activity, whereas no significant difference was observed at 10 mm of NO3–. In the second experiment, carried out under open field conditions, increasing the N fertilization rates from 0 to 120 kg·ha−1 increased the total and marketable yields of melon plants, whereas the NUE decreased. When averaged over N levels, the marketable yield, NUE, and N uptake efficiency were higher by 9%, 11.8%, and 16.3%, respectively, in 'Proteo' grafted onto 'P360' than in ungrafted 'Proteo' plants

Protein Hydrolysates from Animal or Vegetal Sources Affect Morpho-Physiological Traits, Ornamental Quality, Mineral Composition, and Shelf-Life of Chrysanthemum in a Distinctive Manner

Protein hydrolysates (PHs) are a prominent category of plant biostimulants, mainly constituted of amino acids, oligopeptides and polypeptides, obtained by partial hydrolysis of animal or plant protein sources. Despite scientific evidence supporting the biostimulant action of PHs on vegetables, the morphological, physiological, and shelf-life performances underlying the PH action on cut flowers are still poorly explored. Accordingly, the aim of this research is to assess the effects of three commercial biostimulants, one animal PH (PH A, Hicure®) and two plant PHs (PH V1, Trainer®and PH V2, Vegamin©), on two chrysanthemum (Chrysanthemum morifolium) cultivars (Pinacolada and Radost). In both cultivars, only the plant-derived PH (V1 and V2) treatments recorded significantly higher fresh plant biomass than the control (on average +18%, in both cultivars). The foliar application of the vegetal-derived PHs but not the animal one, particularly in Pinacolada, improved the status of plants, stimulating stem elongation and the apical flower diameter. In Pinacolada, applications with PH V1 resulted in a significant increase in nitrate and P concentration in leaves and Ca content in flowers compared with the control (+43%, +27%, and +28% for nitrate, P, and Ca, respectively). In Radost, PH A and PH V2 applications caused a significant reduction in nitrate concentration in both leaves and flowers compared with the control. One week after harvest, in both cultivars, PH A applications caused flower stems to wilt faster than the control. In contrast, plants treated with PH V1 revealed significantly slower flower stem senescence compared to the control. Flower wilting during vase life was correlated to a decrease in the K-to-Na ratio in flowers due to an inability to transport K to the flowers from the leaves rather than an increase in Na in the flowers themselves

Combining Molecular Weight Fractionation and Metabolomics to Elucidate the Bioactivity of Vegetal Protein Hydrolysates in Tomato Plants

The comprehension of the bioactive fractions involved in the biostimulant activity of plant derived protein hydrolysates (PH) is a complex task, but it can also lead to significant improvements in the production of more effective plant biostimulants. The aim of this work is to shed light onto the bioactivity of different PH dialysis fractions (PH1 &lt; 0.5\u20131 kDa; PH2 &gt; 0.5\u20131 kDa; PH3 &lt; 8\u201310 kDa; PH4 &gt; 8\u201310 kDa) of a commercial PH-based biostimulant through a combined in vivo bioassay and metabolomics approach. A first tomato rooting bioassay investigated the auxin-like activity of PH and its fractions, each of them at three nitrogen levels (3, 30, and 300 mg L 121 of N) in comparison with a negative control (water) and a positive control (indole-3-butyric acid, IBA). Thereafter, a second experiment was carried out where metabolomics was applied to elucidate the biochemical changes imposed by the PH and its best performing fraction (both at 300 mg L 121 of N) in comparison to water and IBA. Overall, both the PH and its fractions increased the root length of tomato cuttings, compared to negative control. Moreover, the highest root length was obtained in the treatment PH1 following foliar application. Metabolomics allowed highlighting a response to PH1 that involved changes at phytohormones and secondary metabolite level. Notably, such metabolic reprogramming supported the effect on rooting of tomato cuttings, being shared with the response induced by the positive control IBA. Taken together, the outcome of in vivo assays and metabolomics indicate an auxin-like activity of the selected PH1 fraction

Metabolomic Responses of Maize Shoots and Roots Elicited by Combinatorial Seed Treatments With Microbial and Non-microbial Biostimulants

Microbial and non-microbial plant biostimulants have been successfully used to improve agriculture productivity in a more sustainable manner. Since the mode of action of biostimulants is still largely unknown, the present work aimed at elucidating the morpho-physiological and metabolomic changes occurring in maize (Zea mays L.) leaves and roots following seed treatment with (i) a consortium of two beneficial fungi [arbuscular mycorrhizal fungi (AMF) and Trichoderma koningii TK7] and rhizobacteria, (ii) a protein hydrolyzate-based biostimulant (PH) alone, or (iii) in combination with a consortium of T. koningii TK7 and rhizobacteria. The application of PH alone or in combination with Trichoderma elicited significant increases (+16.6%) in the shoot biomass compared to untreated maize plants, whereas inoculation with AMF + Trichoderma elicited significant increases in root dry biomass (+48.0%) compared to untreated plants. Distinctive metabolomic signatures were achieved from the different treatments, hence suggesting that different molecular processes were involved in the plants response to the biostimulants. The metabolic reprogramming triggered by the treatments including the protein hydrolyzate was hierarchically more pronounced than the application of microorganisms alone. Most of the differential metabolites could be ascribed to the secondary metabolism, with phenylpropanoids and terpenes being the most represented compounds. The application of PH triggered an accumulation of secondary metabolites, whereas the opposite trend of accumulation was seen in the case of microorganisms alone. The increase in biomass could be related to two processes, namely the modulation of the multilayer phytohormone interaction network and a possible increase in nitrogen use efficiency via the GS-GOGAT system

Understanding the biostimulant action of vegetal-derived protein hydrolysates by high-throughput plant phenotyping and metabolomics: A case study on tomato

Designing and developing new biostimulants is a crucial process which requires an accurate testing of the product effects on the morpho-physiological traits of plants and a deep understanding of the mechanism of action of selected products. Product screening approaches using omics technologies have been found to be more efficient and cost effective in finding new biostimulant substances. A screening protocol based on the use of high-throughput phenotyping platform for screening new vegetal-derived protein hydrolysates (PHs) for biostimulant activity followed by a metabolomic analysis to elucidate the mechanism of the most active PHs has been applied on tomato crop. Eight PHs (A–G, I) derived from enzymatic hydrolysis of seed proteins of Leguminosae and Brassicaceae species were foliarly sprayed twice during the trial. A non-ionic surfactant Triton X-100 at 0.1% was also added to the solutions before spraying. A control treatment foliarly sprayed with distilled water containing 0.1% Triton X-100 was also included. Untreated and PH-treated tomato plants were monitored regularly using high-throughput non-invasive imaging technologies. The phenotyping approach we used is based on automated integrative analysis of photosynthetic performance, growth analysis, and color index analysis. The digital biomass of the plants sprayed with PH was generally increased. In particular, the relative growth rate and the growth performance were significantly improved by PHs A and I, respectively, compared to the untreated control plants. Kinetic chlorophyll fluorescence imaging did not allow to differentiate the photosynthetic performance of treated and untreated plants. Finally, MS-based untargeted metabolomics analysis was performed in order to characterize the functional mechanisms of selected PHs. The treatment modulated the multi-layer regulation process that involved the ethylene precursor and polyamines and affected the ROS-mediated signaling pathways. Although further investigation is needed to strengthen our findings, metabolomic data suggest that treated plants experienced a metabolic reprogramming following the application of the tested biostimulants. Nonetheless, our experimental data highlight the potential for combined use of high-throughput phenotyping and metabolomics to facilitate the screening of new substances with biostimulant properties and to provide a morpho-physiological and metabolomic gateway to the mechanisms underlying PHs action on plants