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Rigano, M. M.

Publications and source records attributed to Rigano, M. M..

4 recordsLinked to original sources

Cloudberry-derived nanovesicles: in vitro functional effects in skin cell models and characterization of molecular cargo

Cloudberry (Rubus chamaemorus L.)-derived nanovesicles (NVs) represent a promising but still poorly characterized class of plant-derived vesicles with potential relevance for skin-related applications. Here, we isolated cloudberry fruit-derived NVs and investigated their physicochemical and molecular properties, cellular uptake, cytocompatibility, and functional effects in human dermal fibroblasts (HDF) and HaCaT keratinocytes. Nanoparticle tracking analysis and transmission electron microscopy confirmed a nanosized vesicle preparation with characteristic round morphology, while protein quantification supported reproducible isolation of NV-associated material. In vitro, cloudberry NVs showed concentration-dependent effects on cell viability and proliferation, with lower doses being better tolerated. Labelled NVs were internalized by both HDF and HaCaT cells in a time-dependent manner. Under oxidative stress conditions, cloudberry NVs reduced H2O2-induced senescence-associated {beta}-galactosidase staining in HDFs and exerted cytoprotective effects in both cell lines, alongside measurable cell-free antioxidant activity in the DPPH assay. In scratch wound-healing assays, cloudberry NVs modulated wound closure in a dose-dependent manner, with the lowest tested concentration showing the most favorable response. UHPLC-MS/MS-based proteomics and metabolomics further indicated the presence of diverse secondary metabolites and stress-related protein cargo. Together, these results support the view that cloudberry-derived NVs are biologically active plant nanovesicles with potential utility in skin-related regenerative applications. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=126 SRC="FIGDIR/small/741293v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@c9094dorg.highwire.dtl.DTLVardef@81a1b5org.highwire.dtl.DTLVardef@9fafe2org.highwire.dtl.DTLVardef@1d421d3_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

A tomato line hyposensitive to simulated proximity shade shows altered auxin-related 1 gene expression and improved fruit yield under high-density field conditions

Plants detect the presence of nearby vegetation as a reduction in the ratio of red to far-red light (low R/FR). This proximity shade signal can be simulated in the lab by supplementing white light (W) with FR (W+FR). While shade avoidance strategies are considered undesirable in agricultural crops, FR supplementation enhances plant growth and fruit quality in tomato (Solanum lycopersicum). Here we compared the response of different tomato genotypes to W+FR in the lab and identified one S. pennellii introgression line (IL2-2) with a shade-tolerant phenotype at the seedling stage. Compared to the shade-avoider parental genotype M82, IL2-2 plants showed reduced elongation upon W+FR exposure and a disrupted expression of auxin-related genes both under W and W+FR. At harvest, W+FR treatment improved M82 fruit quality by increasing {degrees}Brix, ascorbic acid and carotenoids, and these quality traits remained virtually unchanged in IL2-2. Under high density (HD) conditions, fruit quality traits were hardly impacted by planting density or genotype, but IL2-2 showed improved fruit yield. Our findings suggest that IL2-2 could serve as a valuable genotype for high-density or intercropping agrosystems.

plant biology↗

Editing strigolactone biosynthesis genes in tomato reveals novel phenotypic effects and highlights D27 as a key target for parasitic weed resistance

Parasitic weed infestations pose an increasing threat to agriculture worldwide, especially in the Mediterranean region. Phelipanche ramosa and P. aegyptiaca (broomrapes) cause severe damage to field-grown tomato (Solanum lycopersicum L.). Strigolactones (SLs), apocarotenoid phytohormones, play a critical role in plant physiology and development, and are also the primary signals that trigger the germination of parasitic weed seeds. We generated CRISPR/Cas9 tomato knock-out lines for the SlD27 gene, as well as three other key genes involved in SL biosynthesis (SlCCD7, SlCCD8, SlMAX1), all within the same genetic background. The edited lines exhibited a marked reduction in SL content in root exudates, along with impaired broomrape seed germination. A comprehensive analysis of morphological, reproductive, and fruit-related traits revealed gene-specific effects on plant phenotype, including vegetative traits, fruit set, fruit development, and volatilome. Specifically, the knock-out of two CCDs and the MAX1 had a specific impact not only on plant development but also on the production of volatile organic compounds during fruit ripening. In contrast, the Sld27 lines, produced for the first time in this study, displayed a phenotype similar to the control non-edited plants, suggesting that the D27 gene holds promise as a breeding target for enhancing resistance to parasitic weeds in tomato. HighlightThe characterization of tomato CRISPR/Cas9-edited lines for the four core genes involved in strigolactone biosynthesis revealed gene-specific effects on plant phenotype, with D27 emerging as a potential target for resistance to parasitic weeds.

genetics↗

Single-cell mass spectrometry reveals heterogeneous triterpenic acid accumulation in apple callus-derived cells

The use of plant cell cultures for large scale production of natural compounds, although promising, has been hindered by their genetic instability and heterogeneity. Here, we show how single cell mass spectrometry can be used to characterize the natural product profile of a callus culture at a highly resolved level. We identify and quantify triterpenic acids in a population of callus cells derived from Annurca apple (Malus pumila Miller cv Annurca) leaf. The analysis demonstrated that a high degree of metabolic heterogeneity exists in the cell population, with the levels of detected metabolites varying significantly across the callus cells. This metabolic heterogeneity was underpinned by variable expression levels of key biosynthetic genes in the single cells. The application of an abiotic stress, near ultraviolet radiation (NUV), to the callus culture resulted in increased levels of triterpenic acids. Single cell mass spectrometry analysis revealed that after treatment, a larger percentage of callus cells produced detectable amounts of these metabolites, ultimately resulting in a more homogeneous production of the metabolites. Furthermore, it showed that intracellular concentrations of ursolic acid derivatives can reach more than 100 mM. Single cell mass spectrometry analyses provide a starting foundation for understanding the molecular mechanisms responsible for metabolic heterogeneity in plant cell cultures, which could in turn facilitate efforts to improve these cell cultures for commercial purposes.

plant biology↗