CONTATTI
PRIVACY & COOOKIE POLICY
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Main Goal of the project
FarmEVs project will develop Extracellular Vesicles-based solutions for human and plant health through the optimization of vegetable-based EV production. It will create innovative methodologies for their characterization and their engineering towards specific applications. To achieve this ambitious objective, FarmEVs project makes use of the complementary expertises of a highly international and inter-sectoral consortium (link to consortium partners).
Objectives:
EV-farm
To isolate plant-derived nanovesicles (PDNVs) from two organic crops: tomato and bean.
To isolate EVs from plant cell cultures.
To develop engineered PDNVs.
Exploitation
To exploit native and engineered plant vesicles in agro-nanotechnological applications, and for their beneficial effects on human health.
Multidisciplinary platform
To setup and apply a multipurpose technological platform for the morphological, physicochemical, molecular, in vitro and in vivo characterization of plant EVs. This will provide solid data sets for basic and applied plant EV studies.
WP1 – The Plant EV Factory
Aim: WP1 will produce and characterize organic PDNVs and will establish a novel EV-farming platform based on plant cell cultures. WP1 is structured into 4 tasks with specific objectives.
EV lab on a farm.
Current adaptive crop management practices as soil conditioning, mulching and biodynamic preparations will be used for the production of organic vegetables. The “EV Lab on Farm” facility that will transform selected organic waste of the farm into a PDNVs enriched biofertilizer will be established. The facility will also be used for demonstration purposes to outreach farmers, industries, scientific communities.
Organic PDNVs.
PDNVs from the homogenates of organically grown vegetables will be isolated at laboratory scale and compared with PDNVs obtained from conventionally grown species.
EV farming: EVs from plant cell cultures.
Cell suspension cultures and hairy root cultures from tomato and bean will be established and EVs will be isolated from the cell culture conditioned medium. The obtained EVs will be characterized and their production will be maintained during the whole lifetime of the project.
Physicochemical and molecular characterization PDNVs and plant EVs using the multidisciplinary platform.
Physicochemical, morphological, molecular, and in vitro characterization of PDNVs and EVs will be performed using a multidisciplinary EV technology approach. Bioinformatics will aid proteomics and transcriptomic data integration and interpretation.
WP2 – Plant EV-based solutions for human and plant health.
Aim: WP2 will produce engineered plant EVs to mitigate plant diseases, promote symbiotic interactions and create high end-value nanodrug-delivery system for human health. WP2 is structured into 4 tasks with specific objectives.
EVs uptake and translocation within whole plants.
The uptake of the vesicles by the root and the leaf cuticle and epidermis as well as their local/systemic transport will be studied in hydroponic cultures of bean and/or tomato plants using fluorescently labeled vesicles.
Vesicle-mediated delivery of small RNAs in tomato plants for inducing virus resistance.
PDNVs expressing small RNAs will be produced for the specific targeting of pathogenic plant viruses, and inducing RNA silencing-mediated viral degradation.
Loading vesicles with exogenous compounds.
Three types of engineered plant vesicles will be produced: anti-inflammatory, antiviral and growth promoting. Loading will be optimized for loading efficiency and capacity. Fluorescent labeling and tracking methods will be used to track subcellular localization. In vitro EV-uptake experiments on L. japonicus plants will be performed using GR24 loaded vesicles in order to estimate their effect on plant-Rhizobium interaction.
In vitro and in vivo assays.
Bioassays of engineered EVs will be performed using human cells in vitro and plants in vivo. PDNVs produced from the organic farm waste at Lehtosarvi farm will be tested for their fertilizing properties in a small-scale experiment.
WP3 – Technological and Methodological Innovations.
WP3 will create and integrate technological and methodological innovations to support the main scientific objectives of FarmEVs project. WP3 is structured into 4 tasks with specific objectives.
A novel microchip-based assay.
A high throughput microfluidic device for ELISA-based anti-inflammatory analysis of PDNVs and plant EVs will be developed. An optical fibre-based “microscale DLS instrument” will be incorporated into the microfluidic device for on-chip analysis.
EVs in plant mutualistic symbiosis.
The temporal and spatial expression profiles of putative Tetraspanin (TSP) sequences that play a role in Lotus japonicus symbiotic interaction will be studied using hydroponic and hybrid (soil-hydroponic) cultures of L. japonicus with/out M. loti inoculation. Quantitative proteomics will be used to determine the taxa distribution and the change in the protein cargo over the time.
Modelling the EV-membrane interactions.
Numerical and Monte Carlo simulation programs for calculation of axisymmetric and non- axisymmetric membrane shapes will be used to study the plasma membrane interactions (encapsulation) with EVs, nanoparticles, bacteria and viruses of different shape, considering the plant cytoskeleton active forces along with the anisotropy of the membrane constituents. The possibility to simulate the adhesion of closed cell membrane shapes to various structured surfaces.
Co-creating agroecological symbioses for the sustainable benefits of horticulture.
Various approaches for the characterization of the symbiosis-driven interactions at different levels, i.e. molecular (proteins) organ (membrane, nodules), organism and macroscopic systems of farming will be evaluated and discussed to the benefit of Agroecological symbiosis.