Foliar applications to vines of methyl jasmonate and nanoparticles doped with methyl jasmonate: impact on grape and wine polysaccharide composition

Polysaccharides in wine play important roles in the stabilization and in the sensory properties of wines. Elicitor application constitutes an interesting field of research since it is indirectly involved in the accumulation in grape cell walls of molecules like callose, lignin, phenolic compounds and glycoproteins. Currently, biomimetic calcium phosphate (ACP) nanoparticles are successfully used in viticulture for the controlled delivery of bioactive molecules, such as elicitors. The aim of this study was to compare the effect of the application of two different elicitors on both grape and wine of Tempranillo polysaccharide composition. Methyl jasmonate (MeJ) and nanoparticles doped with MeJ were applied to the canopy at veraison and one week later in two vintages. In the grape extracts, the foliar treatments did not increase the content of monosaccharides or that of the main pectin families; therefore, the elicitors did not reinforce the cell walls of the Tempranillo grape. The extractability and solubility of the pectic families of the grape cell walls into the wine depended on the type of family and the climate of the vintages.Ministerio de Ciencia, Innovación y Universidades through the Projects RTI2018-096549-B-I00 and RTI-2018-095794- A-C22Ministerio de Ciencia, Innovación y Universidades for her Juan de la Cierva‑Incorporación contrac

Bioinspired mineralization of engineered living materials to promote osteogenic differentiation

In this work, Engineered Living Materials (ELMs), based on the combination of genetically-modified bacteria and mineral-reinforced organic matrices, and endowed with self-healing or regenerative properties and adaptation to specific biological environments were developed. Concretely, we produced ELMs combining human mesenchymal stem cells (hMSCs) and Lactococcus lactis (L. lactis), which was specifically programmed to deliver bone morphogenetic protein (BMP-2) upon external stimulation using nisin, into mineralized alginate matrices. The hybrid organic/inorganic matrix was built through a protocol, inspired by bone mineralization, in which alginate (Alg) assembly and apatite (HA) mineralization occurred simultaneously driven by calcium ions. Chemical composition, structure and reologhical properties of the hybrid 3D matrices were dedicately optimized prior the incorportation of the living entities. Then, the same protocol was reproduced in the presence of hMSC and engineered L. lactis that secrete BMP-2 resulting in 3D hybrid living hydrogels. hMSC viability and osteogenic differentiation in the absence and presence of the bacteria were evaluated by live/dead and quantitative realtime polymerase chain reaction (qPCR) and immunofluorescence assays, respectively. Results demonstrate that these 3D engineered living material support osteogenic differentiation of hMSCs due to the synergistic effect between HA and the growth factors BMP-2 delivered by L. lactisEPSRC (EP/P001114/1)U.K. Regenerative Medicine Platform “Acellular/Smart Materials-3D Architecture” (MR/R015651/1)Junta de Andalucía through the collaborative project NanoFERTI (P18-TP-0969)The Spanish MCIN/AEI/10.13039/501100011033The “European Union” NextGenerationEU/PRTR (project number PDC2022-133191- I00)EMBO Scientific Exchange Grant (SEG number 9741)RYC2021-032734-IMCIN/AEI/10.13039/501100011033ESF Investing in your futur

Effects of Methyl Jasmonate and Nano-Methyl Jasmonate Treatments on Monastrell Wine Volatile Composition

This work has been supported by funding provided by the Spanish MCIN/AEI/10.13039/501100011033 and "ERDF, A way of making Europe" through the projects NanoVIT (RTI-2018-095794-B-C21 and RTI-2018-095794-A-C22), and by the Junta de Andalucia with the project NanoFERTi (P18-TP-969). GBRR also acknowledges Junta de Andalucia for her postdoctoral contract within the PAIDI 2020 program (DOC_01383).The application of methyl jasmonate (MeJ) as an elicitor to enhance secondary metabolites in grapes and wines has been studied, but there is little information about its use in conjunction with nanotechnology and no information about its effects on wine volatile compounds. This led us to study the impact of nanoparticles doped with MeJ (Nano-MeJ, 1mM MeJ) on the volatile composition of Monastrell wines over three seasons, compared with the application of MeJ in a conventional way (10 mM MeJ). The results showed how both treatments enhanced fruity esters in wines regardless of the vintage year, although the increase was more evident when grapes were less ripe. These treatments also achieved these results in 2019 in the cases of 1-propanol, ß-phenyl-ethanol, and methionol, in 2020 in the cases of hexanol and methionol, and in 2021, but only in the case of hexanol. On the other hand, MeJ treatment also increased the terpene fraction, whereas Nano-MeJ, at the applied concentration, did not increase it in any of the seasons. In summary, although not all families of volatile compounds were increased by Nano-MeJ, the Nano-MeJ treatment generally increased the volatile composition to an extent similar to that obtained with MeJ used in a conventional way, but at a 10 times lower dose. Therefore, the use of nanotechnology could be a good option for improving the quality of wines from an aromatic point of view, while reducing the necessary dosage of agrochemicals, in line with more sustainable agricultural practices.Junta de Andalucia P18-TP-969 DOC_01383Spanish MCIN/AEI"ERDF, A way of making Europe" through the projects NanoVIT RTI-2018-095794-B-C21 RTI-2018-095794-A-C2

Nanoparticles doped with methyl jasmonate: foliar application to Monastrell vines under two watering regimes. An alternative to improve grape volatile composition?

Financial support was given by the Ministerio de Ciencia, Innovación y Universidades for projects RTI2018‐096549‐B‐I00 and RTI‐2018‐095794‐A‐C22. EP Pérez‐Álvarez thanks the Ministerio de Ciencia, Innovación y Universidades for her Juan de la Cierva‐Incorporación postdoctoral contract (IJC2019‐040502‐I). GBRR also acknowledges the Junta de Andalucía for her postdoctoral contract within the PAIDI 2020 program (DOC_01383).Background: Elicitors induce defense mechanisms, triggering the synthesis of secondary metabolites. Irrigation has implications for a more sustainable viticulture and for grape composition. The aim was to investigate the influence on grape aroma composition during 2019 and 2020 of the foliar application of amorphous calcium phosphate (ACP) nanoparticles and ACP doped with methyl jasmonate (ACP-MeJ), as an elicitor, with rainfed or regulated deficit irrigation (RDI) grapevines. Results: In both growing seasons, nearly all terpenoids, C13 norisoprenoids, benzenoid compounds and alcohols increased with ACP-MeJ under the RDI regimen. In 2019, under the rainfed regime, ACP treatment increased limonene, p-cymene, α-terpineol, 1,1,6-trimethyl-1,2-dihydronaphthalene (TDN), 2-ethyl-1-hexanol, (E,E)-2,4-heptadienal, and MeJ concentration in comparison with control grapes. In 2020, the rainfed regime treated with ACP-MeJ only increased the nonanoic acid content. Grape volatile compounds were most influenced by season and watering status whereas the foliar application mainly affected the terpenoids. Conclusion: A RDI regime combined with the elicitor ACP-MeJ application could improve the synthesis of certain important volatile compounds, such as p-cymene, linalool, α-terpineol, geranyl acetone, β-ionone, 2-phenylethanol, benzyl alcohol, and nonanoic acid in Monastrell grapes. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.Ministerio de Ciencia, Innovación y Universidades RTI2018‐096549‐B‐I00, RTI‐2018‐095794‐A‐C22, IJC2019‐040502‐I MCIUJunta de Andalucía DOC_0138

Application of Elicitors, as Conventional and Nano Forms, in Viticulture: Effects on Phenolic, Aromatic and Nitrogen Composition of Tempranillo Wines

The phenolic, aromatic and nitrogen composition of a wine determines its organoleptic profile and quality. Elicitors have been used as a tool to stimulate the plant’s defense systems, favoring the synthesis of secondary metabolites. In this pioneering study, the elicitor methyl jasmonate in conventional form (MeJ) and in nanoparticle form (ACP-MeJ), with a concentration ten times lower, was applied in a Tempranillo vineyard over two seasons. The phenolic, nitrogen and volatile composition and the sensory properties of the MeJ-based wines were determined. The results showed that the effects of foliar applications of MeJ modify the wine composition. Thus, although the total concentration of most of the groups of phenolic compounds was not altered, several compounds, such as petunidin-3-glucoside, quercetin-3-glucoside, epigallocatechin and most of the stilbenes, increased, in both years, in the treated wines. Amino acids were influenced differently in each of the years studied, and volatile compounds generally did not improve in the treated wines. However, the ACP-MeJ wines were the best rated by the tasters, highlighting their equilibrium on the taste and their genuineness and odor quality. Therefore, foliar applications of ACP-MeJ can be considered a useful tool to improve wine quality.Spanish Government RTI2018-096549-B-I00 RTI-2018-095794-A-C2

Amorphous vs. nanocrystalline calcium phosphate as efficient nanocarriers of elicitors in vineyards

Nanotechnology is emerging as a potential strategy to achieve sustainable agricultural productivity and global food security. Engineered nanoparticles (NPs) are endowed with the ability to deliver active ingredients in a responsive manner, reducing the adverse environmental impacts in comparison to the conventional practices. However, the relationship between NP features (i.e., size, morphology, surface charge, structure, etc.) and functionality has been scarcely studied so far. In this work, two types of calcium phosphate NPs were functionalized with the plant resistance-inductor methyl jasmonate (MeJ), which stimulates the production of secondary plant metabolites (e.g., anthocyanins, stilbenes, and flavonols) in grapes. The properties of the resulting nanomaterials, namely, elongated apatite (Ap-MeJ) and roundshaped amorphous calcium phosphate NPs (ACP-MeJ), were studied in detail. In addition, the loading capacity, release kinetics and elicitor thermal stability of the two nanosystems were compared. The results indicated that the differences in terms of morphology, crystalline structure and surface charge did not affect the release kinetics nor the protective action offered by the NPs. However, ACP-MeJ showed much higher MeJ loading capacity than Ap-MeJ. Both types of nanomaterials exhibited similar performance in field experiments on Monastrell vineyards (Vitis vinifera L.). Foliar application of ACP-MeJ and Ap-MeJ (1 mM MeJ) produced wines with similar contents of anthocyanins and tannins (around 500 mg L−1 and 1100 mg L−1, respectively), but Ap-MeJ treatment required doubling the NP amount due to its lower MeJ adsorption capacity. Both treatments produced wines with higher tannin concentration than wines from non-treated grapes or wine treated with free MeJ at a much higher dosage (10 mM). Results highlight the potential of ACP and Ap NPs as elicitor nanocarriers enabling enhancement of the quality of wine in a more sustainable manner.Project nanoSOP, PDC2022-133191-I00 funded by MCIN/AEI/10.13039/ 501100011033The “European Union NextGenerationEU/PRTR” and projects NanoVIT, RTI- 2018-095794-A-C22RTI-2018-095794-B-C21 funded by MCIN/AEI/10.13039/501100011033ERDF A way of making Europe”. JMDLRYC-2016-21042 and RYC2021-032734MCIN/AEI/10.13039/501100011033“ESF Investing in your future”. GBRR also acknowledges Junta de Andalucia for her postdoctoral contract within the PAIDI 2020 program (DOC_01383)

Multifunctional Nanomaterials for Biofortification and Protection of Tomato Plants

Calcium phosphate nanoparticles were doped with zinc ions to produce multifunctional nanomaterials for efficient agronomic fortification and protection of plants. The resulting round-shaped nanoparticles (nanoZn) were composed of 20.3 wt % Ca, 14.8 wt % P, and 13.4 wt % Zn and showed a pH-controlled solubility. NanoZn were stable in aqueous solutions at neutral pH but dissolved in citric acid at pH 4.5 (i.e., the pH inside tomato fruits), producing a pH-responsive delivery of the essential nutrients Ca, P, and Zn. In fact, the foliar application of nanoZn on tomato plants provided tomatoes with the highest Zn, Ca, and P contents (causing, respectively, a 65, 65, and 15% increase with respect to a conventional treatment with ZnSO4) and the highest yields. Additionally, nanoZn (100 ppm of Zn) inhibited in vitro the growth of Pseudomonas syringae (Ps), the main cause of bacterial speck, and significantly reduced Ps incidence and mortality in tomato seeds, previously inoculated with the pathogen. Therefore, nanoZn present dual agricultural applicability, enriching crops with nutrients with important metabolic functions in humans and simultaneously protecting the plants against important bacterial-based diseases, with considerable negative impact in crop productionJunta de Andalucía through the project NanoFERTI (P18-TP- 969)Projects NanoFERTI2 (CPP2021-008801)NextGenerationEU/PRTRRYC2021-032734-I funded by MCIN/AEI/10.13039/501100011033European Union NextGenerationEU/PRT

Organic/inorganic hydrogels by simultaneous self-assembly and mineralization of aromatic short-peptides

This study was supported by projects RTI-2018-095794-A-C22, RYC-2016-21042 and project PID2020-118498GB-I00 funded by MCIN/AEI/10.13039/501100011033 and projects P18-FR-3533 and P18-TP-0969 by FEDER/Junta de Andalucia-Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades (Spain). Thanks go to the CIC personnel of the University of Granada for technical assistance. MCMT acknowledges grant PRE2018-083773 funded by MCIN/AEI/10.13039/501100011033 and FSE "El FSE invierte en tu futuro", Spain. GBRR also acknowledges Junta de Andalucia for her postdoctoral contract within the PAIDI 2020 program ( DOC_01383).Self-assembled peptides and proteins have turned out to be excellent templates for the growth of inorganic minerals and can be used to emulate natural biomineralization processes. Doing this, researchers have developed complex sophisticated materials with properties, in some cases, similar to those found in nature. Of special interest is the development of scaffolds able to guide bone regeneration. The bone tissue comprises an organic matrix composed of aligned collagen fibers containing nanoapatite crystals oriented along the fiber direction. During bone mineralization, both processes, the self-assembly of collagen fibrils and mineralization occur simultaneously. Collagen fibers are able to control calcium phosphate nucleation and subsequent apatite crystal growth at a very limited range of collagen density and ionic concentration. In this study, we reproduced the simultaneity of both processes using an artificial peptide fluorenylmethoxycarbonyl-diphenylalanine (Fmoc-FF) that has the ability to self-assemble in water after the addition of Ca2+ ions. Therefore, the peptide self-assembly process and the mineralization of apatite are Ca-demanding processes and occur simultaneously. The role of peptide and ionic concentrations has been investigated affording organic/inorganic hybrid hydrogels with different degrees of homogeneity and mineralization. Interestingly, at very low Ca2+ concentrations, we found that apatite nanocrystals are integrated into Fmoc-FF fibrils and oriented as in biologically mineralized collagen fibrils, the basic building blocks of bone.FEDER/Junta de Andalucia-Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades (Spain) P18-FR-3533 P18-TP-0969FSE "El FSE invierte en tu futuro", SpainJunta de Andalucia DOC_01383; RTI-2018-095794-A-C22; RYC-2016-21042; PID2020-118498GB-I00; MCIN/AEI/10.13039/501100011033; PRE2018-08377