Periodic Reporting for period 4 - NoPest (Novel Pesticides for a Sustainable Agriculture)
Reporting period: 2023-07-01 to 2024-12-31
NoPEST – Novel PESTicides for a Sustainable Agriculture – is a five-year project funded under the FET-OPEN “Challenging Current Thinking” programme. Its goal is to pioneer innovative, low-risk and environmentally friendly solutions for crop protection, contributing to a more sustainable agricultural system in Europe and beyond. Modern agriculture still relies heavily on chemical pesticides, whose global use has increased by 15–20 times in recent decades, causing environmental pollution, greenhouse gas emissions, and health risks. In response, the EU (Directives 1107/2009/EC and 473/2002) has tightened pesticide regulation, creating an urgent need for safer alternatives.
NoPEST addresses this challenge by developing next-generation plant protection tools based on peptide aptamers – short, high-affinity molecules that inhibit specific target enzymes. These peptides are composed of natural amino acids, have minimal impact on health and the environment, and can be tailored to target virtually any plant pathogen. Precision agriculture technologies are integrated to optimise application timing and location, reducing costs and environmental impact. The project targets oomycete pathogens such as Phytophthora infestans, P. capsici, and P. viticola, which threaten food security and biodiversity. Current control relies on highly polluting copper-based compounds. NoPEST proposes safer alternatives through a multi-step approach: identifying key cell wall biosynthesis enzymes, generating and screening peptide libraries, selecting active molecules, optimising their structure, and developing smart deployment systems.
NoPEST addresses this challenge by developing next-generation plant protection tools based on peptide aptamers – short, high-affinity molecules that inhibit specific target enzymes. These peptides are composed of natural amino acids, have minimal impact on health and the environment, and can be tailored to target virtually any plant pathogen. Precision agriculture technologies are integrated to optimise application timing and location, reducing costs and environmental impact. The project targets oomycete pathogens such as Phytophthora infestans, P. capsici, and P. viticola, which threaten food security and biodiversity. Current control relies on highly polluting copper-based compounds. NoPEST proposes safer alternatives through a multi-step approach: identifying key cell wall biosynthesis enzymes, generating and screening peptide libraries, selecting active molecules, optimising their structure, and developing smart deployment systems.
NoPEST made significant progress towards the development of innovative, sustainable solutions for crop protection based on peptide aptamers, peptidomimetics, non-peptide small molecules, and precision agriculture technologies.
1. Target Identification and Enzyme Characterization (WP1)
Efforts focused on elucidating the structure and biosynthesis of oomycete cell wall polysaccharides as novel targets for disease control. Ten key enzymes from Phytophthora infestans, P. capsici, and P. viticola were heterologously expressed and purified in various host systems (S. cerevisiae, P. pastoris, E. coli). Biochemical assays confirmed the activity of several glycosyltransferases and effector proteins, and their susceptibility to selected peptide aptamers was validated. These results provided essential targets for downstream aptamer screening in WP2.
2. Peptide and Peptidomimetic Optimization
Extensive work on the design and synthesis of LF20 and LF32 peptide analogues was carried out. Chemical modifications, including the introduction of D-amino acids, hydrazino acids, and β-amino acids, enhanced peptide stability against proteolysis without compromising antimicrobial activity. Recruitment of dedicated researchers at partner institutions supported Ala-scan analyses and structural optimization efforts. Peptidomimetics derived from CP20 and CP32 showed improved antimicrobial activity in vitro and ex vivo and demonstrated good ecotoxicological profiles, with only minor phytotoxic effects observed in some cases.
3. Non-Peptide Small Molecules
Refined pharmacophore modelling identified new small-molecule candidates with improved antimicrobial efficacy and no observed toxicity to non-target organisms or plants. One lead compound with >70% protective activity was selected for further optimisation, potentially in collaboration with industrial partners.
4. Precision Agriculture and AI Tools
Advanced sensing and imaging technologies were deployed, including manual, mobile, and drone-based RGB imaging, leading to the acquisition of over 2,500 field images. AI-based models (CNNs, YOLOv4, SegNet) achieved high accuracy in detecting downy mildew and late blight, enabling precise disease mapping and variable rate application (VRA) strategies.
5. Formulation, Application, and Field Validation (WP8)
Lead peptide (CP20, LP20) were formulated and tested under greenhouse and field conditions across Italy, Spain, and France. Their mobility within plant tissues was assessed, revealing limited systemic translocation. A comprehensive cost-benefit analysis (CBA) compared new biopesticides and precision application strategies with conventional practices demonstrated potential economic, environmental, and societal benefits for European agriculture.
Overall, NoPEST successfully achieved key scientific and technical milestones, advancing from molecular target identification to field validation of two novel peptide-based plant protection solutions. These results have been patented (WO2025093718) and they represent an important contribution to sustainable crop protection and the development of next-generation, environmentally friendly alternatives to conventional pesticides.
1. Target Identification and Enzyme Characterization (WP1)
Efforts focused on elucidating the structure and biosynthesis of oomycete cell wall polysaccharides as novel targets for disease control. Ten key enzymes from Phytophthora infestans, P. capsici, and P. viticola were heterologously expressed and purified in various host systems (S. cerevisiae, P. pastoris, E. coli). Biochemical assays confirmed the activity of several glycosyltransferases and effector proteins, and their susceptibility to selected peptide aptamers was validated. These results provided essential targets for downstream aptamer screening in WP2.
2. Peptide and Peptidomimetic Optimization
Extensive work on the design and synthesis of LF20 and LF32 peptide analogues was carried out. Chemical modifications, including the introduction of D-amino acids, hydrazino acids, and β-amino acids, enhanced peptide stability against proteolysis without compromising antimicrobial activity. Recruitment of dedicated researchers at partner institutions supported Ala-scan analyses and structural optimization efforts. Peptidomimetics derived from CP20 and CP32 showed improved antimicrobial activity in vitro and ex vivo and demonstrated good ecotoxicological profiles, with only minor phytotoxic effects observed in some cases.
3. Non-Peptide Small Molecules
Refined pharmacophore modelling identified new small-molecule candidates with improved antimicrobial efficacy and no observed toxicity to non-target organisms or plants. One lead compound with >70% protective activity was selected for further optimisation, potentially in collaboration with industrial partners.
4. Precision Agriculture and AI Tools
Advanced sensing and imaging technologies were deployed, including manual, mobile, and drone-based RGB imaging, leading to the acquisition of over 2,500 field images. AI-based models (CNNs, YOLOv4, SegNet) achieved high accuracy in detecting downy mildew and late blight, enabling precise disease mapping and variable rate application (VRA) strategies.
5. Formulation, Application, and Field Validation (WP8)
Lead peptide (CP20, LP20) were formulated and tested under greenhouse and field conditions across Italy, Spain, and France. Their mobility within plant tissues was assessed, revealing limited systemic translocation. A comprehensive cost-benefit analysis (CBA) compared new biopesticides and precision application strategies with conventional practices demonstrated potential economic, environmental, and societal benefits for European agriculture.
Overall, NoPEST successfully achieved key scientific and technical milestones, advancing from molecular target identification to field validation of two novel peptide-based plant protection solutions. These results have been patented (WO2025093718) and they represent an important contribution to sustainable crop protection and the development of next-generation, environmentally friendly alternatives to conventional pesticides.
NoPEST achieved all scientific and technical objectives, advancing from molecular discovery to field validation of two novel peptide-based solutions. These results have been patented (WO2025093718), representing a major step toward sustainable plant protection. The project not only met but in many aspects exceeded its initial goals. The resulting intellectual property is jointly owned by University of Milan (UMIL), KTH Royal Institute of Technology, and Université Paris-Saclay (UPSaclay). A patent covering the antimicrobial peptides has been licensed to SIPCAM-OXON, which aims to scale up production via fermentation-based processes to reduce costs. The technology’s commercial potential has attracted significant industrial interest, with several agri-tech companies initiating collaborative research contracts with UMIL. In parallel, UMIL is finalising the Proof-of-Concept phase of the spin-off PeptoFarm (https://peptofarm.it/en/(opens in new window) designed to accelerate market deployment of peptide-based crop protection technologies. Negotiations with venture capital investors are underway to support its launch and growth. Beyond antimicrobial applications, the NoPEST platform is now being explored for the identification of herbicidal and biostimulant peptides, demonstrating its versatility, scalability, and potential to transform multiple areas of sustainable agriculture.