The partners

GBF team has a world-wide recognition in the field of fruit research using tomato as a model system and has made a substantial contribution to the generation of genomics resource on the tomato model. It has been leading the French contribution to the tomato genome sequencing project. In the last period, GBF team identified a master regulator of fruit texture allowing to uncouple fruit ripening from fruit softening thereby bringing a paradigm shift of thinking. GBF also made a major contribution to unravelling the role of ethylene and auxin in tomato fruit development and ripening. Because of its expertise on fruit ripening, GBF will particularly participate to the definition of the target of new bioactive molecules in this project. The GBF routinely implements combined functional genomics and reverse genetics approaches to uncover the physiological significance of regulatory genes, which will also contribute to the project’s objectives.

GBF largely contributed to the generation and assembly of the tomato genome sequence using NextGen sequencing technologies and has been involved in major EU initiatives dealing with fruit development, quality and ripening such as the COST RoxyCOST chaired by Julien Pirrello (2019-2024), QualityFruit European network (2012-2016), TomGEM H2020 project (2016-2020), HARNESSTOM H2020 project. JP is also PI of the National Funded Project OxyFruit (ANR: 2023-2027). 

The group has a long-term expertise and world-wide recognition in molecular plant hormone physiology and vitamin biology. In addition to the physiological expertise, UGent-FPB has strong expertise in plant histology and bio-analytics of small molecules (particularly as part of ethylene and vitamin metabolism) by mass-spectrometry and photoacoustics-based technologies. This expertise will be an asset to WP2 (identifying small molecules affecting ethylene metabolism), WP3 (impact of different storage methodologies on nutritional quality, assessing impact B-vitamin accumulation), and WP4 (unraveling molecular mechanisms underlying the action of identified molecules, with particular expertise on ethylene metabolism, and tissue-specific effects of compounds through histological analysis).

The Plant Energy Biology Lab will deliver expertise in in vivo monitoring of metabolites and signalling compounds by fluorescent protein biosensors (SO2). It has developed a comprehensive platform of genetic resources, including sensor constructs, overexpression clones, and Arabidopsis lines. The combination of those resources and associated protocols are unique for monitoring a wide range of cell-physiological parameters such as glutathione redox potential, H₂O₂, pH, Ca²⁺, ATP, NAD⁺/NADH ratio, and NADP⁺/NADPH ratio. The lab employs confocal laser scanning microscopy for high resolution biosensing at low throughput, and plate-reader-based fluorimetry for low resolution biosensing at high throughput. A unique feature of both setups is the ability to adjust experimental conditions, such as oxygen concentration as well as chemical treatments live during the measurement, in order to follow responses of multiple live tissue samples in parallel (paraplexing of different biosensors and treatments). This allows performing experiments live on the microscopy stage or in the 96-well plate. For the exposure of live plant samples to changing environmental conditions, the imaging equipment has been extended with custom instrumentation to adjust oxygen concentrations by nitrogen flushing as well as injection or perfusion with solutions to exchange treatment chemicals. Those flexible setups will be exquisitely suited to the objectives on this consortium, where they will be deployed for the investigation of the impact of small molecule effects on stress responses and signalling. The biosensing approach has been instrumental in constructing physiological maps of the plant cell and elucidating compartment-specific dynamics and regulation. The methodology allows for testing specific hypotheses about the control of subcellular physiology and metabolism but also screening for the impact of chemicals on subcellular physiology. Recent publications from the lab underscore its commitment to mechanistic rigor at both technical and biological levels. Having its focus on fundamental research using Arabidopsis as its main model, the lab has been involved in a number of national and international collaborations and research consortia and is experienced in training external scientists. This innovative methodological spectrum positions the lab to make significant contributions to SO2 and SO3 to identifying and validating novel chemical compounds to manipulate pathways of fruit ripening, with a particular focus on cold, hypoxia and ROS signalling.

By applying molecules identified in WP2, Akdeniz university will examine their effects on fruit and vegetable quality during storage and shelf life. They will compare the efficiency of these new identified molecules with existing 1-MCP and low oxygen applications. Colour change, fruit firmness, SSC, TA, lycopene, total phenolics, carotenoids, vitamin c will be the main characters assessed to determine the main effects of these applications on fruit quality. They will organize training school on “Regulation of ripening during postharvest fruits” and a “Molecules role in fruits ripening and its practical applications in industrial scale”.

Genetically engineered auto luminescence as the basis of “glowing” plant and fruit sentinels will be an innovative and valuable tool to remotely assessed endogenous and/or environmental cues . The cue sensors will be promoters from genes responding to the stimulus of interest that will be used to drive the expression of one selected genes in the bioluminescence cycle in sentinel plant that contains the rest of the bioluminescence genes under the control of the 35S promoter. Selected promoters to test will be a promoter from Pathogenesis Related genes responding to different types of stress, promoters responding to hypoxia and cold stress will be also considered. Also synthetic promoters and native promoters that respond to ABA, Ethylene and Auxin will be used in single cue sensors.  This auto luminescent plant is an innovative and powerful material to study the role of different molecules on fruit ripening. This is to be developed in Arabidopsis, tobacco and tomato in order to produce suitable plant material to assess the impact of natural bioactive molecule in High-Throughput assay.

This group has broad expertise in molecular biology, plant genetic transformation, transcriptomics, metabolomics, metagenomics, bioinformatics, with particular regard to gene/metabolite correlation-based analysis. They will participate in WP3, using pharmacological treatment (by spraying) with carotenoids and apocarotenoids. They will test efficiency of these bio active molecule on post-harvest shelf life. Also, they will characterize the post-harvest ripening through metabolomics analysis. In WP4 they will characterize samples subjected to treatment with bioactive compounds at metabolomics levels. As expert in carotenoid and apocarotenoid metabolomics pathway, BIOTEC group will pay particular attention to this pathway, including the precursor of phytohormones.

Gulbudak is a major actor in the post-harvest sectors. They have the highest cold room in Turkey for apple storage and they invest in 2 big cold storage with DCA technology. As a major actor in the sector, they care about renewable energy systems and aiming for maximum efficiency in energy use. Their facilities consisting of 2 pack houses and 103 cold rooms is using electric energy from the solar panels since 2020. In the WP3 they will contribute to assess the efficiency of new bioactive molecules in post-harvest storage shelf life. In addition, their long experience in post-harvest storage, their input in WP5 will be important to communicate and disseminate the results to the main stakeholders. Then, they will be involved in the organization of training school and workshop related to the use of low oxygen in fruit physiology and post-harvest storage.

CLOUD is an SME specializing in cloud-based drug design and pharmaceutical R&D. In WP2, CLOUD will perform a structure-based virtual screening protocol to identify bioactive molecules from their in-house web platform CNatural (http://cnatural.gr/home/), which is focusing on chemical and biological information of naturally occurring compounds (currently ~200,000 molecules) gathering relevant data from several public sources. The primary objective will be identification of novel bioactive molecules interacting with enzymes involved in the enzymatic regulation of fruit ripening. In addition, given their vast experience in developing artificial intelligence models able to predict the toxicity of chemical compounds (ENVIROMED), their input in WP2 will be crucial for the evaluation of potential side effects of the molecules under study.

The PlantLab will deliver expertise in

(1) Establishment of the High-Throughput System platform to optimize growth conditions.

(2) Implement a bioassay to identify molecules from the tested set that can induce a detectable phenotypic effect in treated plants, leading to significant variations in selected biometric parameters.

(3) Molecular characterization of the tested compounds based on the specific effects observed.

(4) In-depth investigation into the mechanisms underlying their mode of action.

The initial screening relies on a visual analysis of the phenotype, employing the following instruments depending on this screening system and the plant age used for the test: LemnaTec 3D phenotyping machine, used to monitor the trends of biometric parameters (digital biomass, green index, Leaf area index) in pots of Arabidopsis plants. The screening will rely on following assay will be performed:

• Wide-field fluorescent microscope (Leica THUNDER imager) to analyse plant growth in 96-well plates system.
• Night Shade In Vivo Plant Imaging System (Berthold) to acquire molecular images with report line of Arabidopsis seedlings.
• Luminometer (Berthold), multimode microplate readers to measure the Relative Luminescence Unit (RLU) in report line of Arabidopsis seedlings.

Isolcell will contribute to the application of the selected biomolecules and to test their efficiency in post-harvest commercial storage room (WP4). They will characterize the ripening of fruit during post-harvest storage through the measurement of different parameters such as respiration rate, ethylene measurement. In addition, they will assess the effect of ethylene removal, low oxygen and CA on the storability of fruit and vegetables. The shelf life can be altered by development of superficial fungi. Isolcell company will test the effect of ROS on fungi development.

The group focuses on tomato developmental processes, including fruit ripening, using molecular biology approaches including tomato agrobacterium mediated transformation, transient and stable with emphasis on prolyl-4-hydroxylases, cell wall hydroxyproline rich glycoproteins, hormone peptides. The group will use the hypoxylab (Oxford optronix) to assess the effectiveness of molecules in different oxygen concentrations, imposed with outmost precision, in tomato protoplasts. Their effect on the molecular mechanisms of fruit ripening under low oxygen conditions (fruit tissue frozen in liquid nitrogen under low oxygen conditions without exposure to atmospheric conditions) will be determined using cell wall glycoproteins characterization. Moreover, the group currently develops the use of an in vitro fruit ripening system in microtome tomato transformed with a specific gene construct. Microtome tomato fruits grow directly from callus after agrobacterium-mediated transformation which can be used as a semi-in vivo system to characterize fruit ripening in response to bioactive molecules, complementing the tomato protoplast approach.

CTIFL is a French national network for applied research aiming at answering fruit and vegetable growers’ questions and finding solutions to their needs. A new postharvest and quality platform of CTIFL Saint-Rémy-de-Provence was built in 2021. The 1700 m² - warehouse has 35 storage rooms of different sizes (85-95m³ to 0.20m³) with a large range of technologies (air-forced tunnel, normal cold, CA and XULO (eXtreme Ultra Low Oxygen) storage rooms, ripening rooms (CA + Thermal Treatment with high CO2 for insect sanitation) with computer supervision. These facilities will be used to test the impact of bioactive molecules on post-harvest storage (WP3). Using their skills in storage management and equipment and fruit quality control, they will provide complete analysis of the produce: microbiology; quality (physical and biochemical parameters, nutritional and volatiles compounds). In addition, CTIFL has developed a range of services, professional tools and training courses as well as publications in various formats to transfer innovative practices and provide the best support possible to the stakeholders in the sector. CTIFL organizes Technical Meetings and Webminars on Storage and participate to Scientific and professional Symposia or TradeShow (INTERPOMA, SIVAL, ISHS, Post-Harvest Unlimited, FRUITLOGISTICA…). Consequently, its role in dissemination and communication to the different stakeholder through organization of Technical Meetings and Webminars on Storage, will be overriding (WP5).