Deliverable D4.4 - Guidelines to implement novel breeding techniques and approaches

Page 1/35 DIVINFOOD Co-constructing interactive short and mid-tier food chains to value agrobiodiversity in healthy plant-based food Deliverable D4.4 Guidelines to implement novel breeding techniques and approaches Due date of deliverable: M42 Actual submission date: M42 Start date of the project: March 1st, 2022 Duration: 60 months Organisation name of lead contractor: Research Institute of Organic Agriculture FiBL Revision: V4 Dissemination level Public – PU X Confidential, only for members of the consortium (including Commission Services) – CO Classified, as referred to in Commission Decision 2001/844/EC - Cl Page 2/35 Metadata CALL H2020-SFS-2020-2 Topic SFS-01-2018-2019-2020 - Biodiversity in action: across farmland and the value chain Project ID 101000383 Project website www.DIVINFOOD.eu Document type Deliverable Title Deliverable D4.4 - Guidelines to implement novel breeding techniques and approaches Author(s) Mariateresa Lazzaro (FiBL), Verena Simon-Kutscher (FiBL) Date of creation 29.08.2025 Version number V4 Internal reviewers Francesca Galli (UniPi), Dalia Mattioni (UniPi), Yuna Chiffoleau (INRAE), Vincent Troillard (INRAE) Keywords seed production, cultivar testing, breeding, molecular breeding, participatory breeding, multi-actor involvement, Neglected and Underutilised Crops (NUCs), social innovation Dissemination level Public To cite this document: Lazzaro M, Simon-Kutscher V, 2025. Deliverable D4.4 - Guidelines to implement novel breeding techniques and approaches. DIVINFOOD H2020 project, report, 36 p. 10.5281/zenodo.17008686 Page 3/35 Table of contents 1. From the need of seed diversity to the opportunity to innovate the breeding sector ... 5 2. Guidelines for fostering NUCs novel breeding techniques and approaches ...................... 6 2.1. From genetic resources management to new cultivar development ................................. 9 2.2. Dynamic Management of Genetic Resources ............................................................................ 12 2.3. Active feedback from cultivation to conservation .................................................................. 13 2.4. Ensuring Seed Quality & Availability ............................................................................................ 14 2.5. Foster technical innovation in NUCs breeding ......................................................................... 15 2.6. More breeding for complex cropping systems ......................................................................... 16 2.7. Breeding for Diversity ......................................................................................................................... 17 2.8. Align breeding with the ethical values of involved stakeholders ..................................... 17 2.9. Promoting Access to Seeds ............................................................................................................... 18 2.10. Multi-Actor involvement .................................................................................................................... 19 2.11. Goals setting in Multi-Actor Breeding .......................................................................................... 21 2.12. Involvement of consumers and citizens ...................................................................................... 25 3. References .............................................................................................................................................. 28 3. Appendixes ............................................................................................................................................. 29 Page 4/35 Executive summary The Deliverable 4.4 presents Guidelines (related to materials, techniques, approaches and collaborations) to implement novel breeding techniques and approaches in the Neglected and Underutilised Crops (NUCs) seed and breeding sector. The recommendations listed in this report have emerged from the work of the DIVINFOOD Living Labs (LLs) with minor cereals and legumes in various regions of Europe. The target audience for this report is stakeholders, including breeders, researchers, farmers and network facilitators interested in developing the seed and breeding sector of minor crops or open to explore alternative paths in their work in major crops. Although this report is based on the specific experiences of the DIVINFOOD LLs on advancing cultivation and use of Neglected and Underutilised Crops (NUCs), the guidelines presented can serve as reflection points for any network working on seeds with a specific interest in the overall sustainability of the food system. Authors: Mariateresa Lazzaro (FiBL) &Verena Simon-Kutscher (FiBL) Internal reviewers: Francesca Galli (UniPi), Dalia Mattioni (UniPi), Yuna Chiffoleau (INRAE), Vincent Troillard (INRAE) Acknowledgements: All members of the DIVINFOOD LLs that contributed to the discussion on the seed and breeding aspects of their network. Page 5/35 1. From the need of seed diversity to the opportunity to innovate the breeding sector The current agri-food system in Europe is based high-input agriculture, reliant on synthetic fertilizers and pesticides, with large scale use of monocrop and limited number of crops in rotation per farm. Agricultural biodiversity has strongly declined in the last century, and it is still threatened because of the industrialisation and market concentration in the agriculture and food sector. Currently, only 150 to 200 of the 250,000 to 300,000 known crops are used by humans. Just three major crops (rice, maize and wheat) provide nearly 60 percent of the calories and proteins humans get from plants. In this general context, the move towards greater agrobiodiversity on farms faces various constraints, including a lack of seeds and cultivars for other species, particularly the neglected and underutilised crops (NUCs). The DIVINFOOD project aims to increase the use of NUCs in Europe at all levels of the value chain, from consumption and food production to farming practices, including seed production and cultivar choice. However, this project seizes the opportunity presented by the lack of NUCs seeds, in terms of both quantity and quality, to propose novel breeding approaches and techniques, contributing not only to the supply of seeds but also to the transformation of the seed and breeding sector, so that it takes greater account of the multiple challenges associated with sustainability and of a larger range of stakeholders and expectations. Figure 1. DIVINFOOD “from plate to seed " approach, in which NUCs are valued, in each LL, via interconnected value chains from food consumption (orange), along the value chain (blue), including breeding, cultivar choice and seed production (green) (source: authors) Adopting a "from plate to seed " approach (Figure 1), which means developing value chains that address the issue of seed availability and cultivar choice beyond the consumption and cultivation step, DIVINFOOD contributes to advance minor crops seed production, to expand cultivar choice and boost new cultivar development (i.e. breeding). The seed production activities aim to provide farmers with high quality seeds of minor crops for inclusion in environmentally and economically valuable crop production. The cultivar selection and breeding activities aim to identify and develop more performing cultivars of minor cereals and legumes with local adaptation, intraspecific biodiversity, and tolerance to biotic and abiotic stress, while also promoting nutritious and tasty food. DIVINFOOD Living Labs (LLs, Appendix I) explore novel breeding techniques and approaches to bring innovation in the seed and breeding sector in Europe with focus on minor crops, and a more general intention to inspire change in approach in this sector in Europe. Page 6/35 2. Guidelines for fostering NUCs novel breeding techniques and approaches To increase cultivation and use of minor crops, it is necessary that farmers can access an adequate quantity and quality of seeds, and that adapted cultivars for different cultivation and consumption environments are developed, considering environmental and management diversity, as well as nutritional and socio-cultural aspects. Such a task requires a multifaceted approach, addressing the key domains of technical and social innovation in breeding, cultivar testing and seed production (Figure 2), considering the linked aspects in the crop production, food processing and consumption domains of the food system. In fact, these guidelines aim to support breeding initiatives that want to deliver seeds suitable for agroecology and organic oriented farming system and whose produce fits in healthy and diverse diets. They are inspired by the DIVINFOOD Living Labs activities in building resilient cultivars and quality seeds for minor crops, as part of their wider aim to increase the sustainability of: (i) crop production by promoting crop diversification and the application of agroecological practices, as well as facilitating the delivery of ecosystem services; (ii) food processing and consumption by promoting short value chains, mild processing and the production of diverse, tasty and nutritious food. Figure 2. Key aspects of Seeds & Breeding sector to consider when working with minor crops to be included in sustainable farming and food production and consumption (source: authors). For minor crops, access to adapted cultivars and seeds in suitable quantities and of appropriate quality is often a major bottleneck for value chain development. Seed & Breeding development goes hand in hand with improving farming practices, developing food products and their marketing. Page 7/35 When working within neglected crops, the specific characteristics of these crops compared with major staple foods and commodity market crops must be carefully considered. Often one needs to apply different strategies compared to similar work with the major crops. Minor crops may have specific agronomic and cultural qualities, such as visual appearance, taste, and processing properties. These qualities must be taken into account for the choice of breeding and selection techniques and approaches. Yet minor crops are also a playground for implementation of novel breeding techniques and approaches with potential of upscale in major crops as well. We present general guidelines related to breeding, cultivar testing and seed production of minor crops, grouped for the key aspects of materials, techniques, approaches and collaborations (Figure 2). The learnings and experiences of DIVINFOOD LLs, both in terms of technical and social innovation, are summarised to inspire breeders, researchers and farmers who intend to work with minor crops. Obviously, these guidelines are not exhaustive or complete and they should be read in context. They should be evaluated and considered in relation to the crop in question, the target region and the relevant value chain. The recommendations consider that, when working on minor crop breeding and seed production, a first domain of decision-making revolves around which MATERIALS to prioritise. This involves, for example, evaluating whether initiating a dedicated breeding programme would be economically and strategically viable, or whether dynamic management of existing genetic resources would be effective enough— particularly for crops with very limited commercial potential, but high local relevance. It also involves considering the opportunities to work with landraces in traditional growing regions, depending on the target market. Similarly, it involves deciding which materials to test first in new growing regions, where a NUC is not traditionally used, once a NUC is of interest for specific opportunities. A second issue to consider is which TECHNIQUES are appropriate for the aim of the program. Leveraging molecular breeding1 for NUCs improvement is strategic, especially in the context of open access data framework, which allows exploitation of genomic resources (e.g. sequencing data and DNA markers) by small scale and independent breeding initiatives that do not have own development capacity. Recent breakthroughs in genomics, can be instrumental in enhancing the genetic potential of minor crops. When molecular data are made available to everyone in open access, this facilitates the application of advanced breeding strategies and technologies. Phenotyping methodologies and platforms specific for the traits of interest in minor crops are also fundamental and very often missing. The third domain concerns the APPROACHES to cultivar development, where decisions must be made, for example, between producing homogeneous varieties—often preferred for yield potential and standardized processing—or heterogeneous populations, which offer greater resilience and adaptability, especially valuable for minor crops grown in diverse and challenging environments. The fourth and equally vital domain is related to who is be involved, meaning the COLLABORATIONS across the value chain. For neglected crops, building a functional value chain frequently requires multiactor collaboration, integrating farmers, researchers, processors, and consumers. Participatory 1 Molecular breeding is the application of molecular biology tools in cultivar development. Here we refer to marker-assisted selection (MAS), as a technique that utilizes DNA markers closely linked to phenotypic traits to enhance selection schemes for specific breeding objectives. Molecular breeding includes the identification and characterization of genetic markers to improve crop traits and facilitate genetic improvement. Page 8/35 breeding approaches are often essential here, as they ensure that crop improvement aligns with local needs while simultaneously fostering the development of a supportive ecosystem around these crops. Additionally, when looking at seed production the inherently low volume of production for many NUCs severely limits the economic viability and applicability of conventional business and technological approaches within the formal seed market. There is a pressing need for improved seed sector (Figure 3) capable of handling and distributing smaller quantities of seed while rigorously maintaining seed quality. An integrated approach for NUCs, which strategically supports and links both informal farmers’ seed networks and formal seed sector, is an opportunity for non-profit and SMEs organisations. Figure 3. Representation of a sustainable seed sector, based on the diversity of their actors and a facilitated flows of germplasm and knowledge amongst them, in which the formal and informal domains are equally valued and interact in a positive way (source: DIVERSIFOOD project Booklet 4). Page 9/35 MATERIALS 2.1. From genetic resources management to new cultivar development An important decision to be made when working to provide short value chains with seeds and cultivar choice for underutilised crops is positioning the envisioned activity from working with genetic resources to new breeding activities. In general terms, the range of methods to maintain and increase genetic diversity and select and develop suitable cultivars spans from Genetic resources Dynamic Management to Development of new varieties (Figure 5, upper axis). The range goes from working with one genetic resource (e. g. the valorization in a local value chain of a traditional landrace for a certain neglected crop), to cultivar testing of already available materials (not used any more or newly brought in from a cultivation region) up to the creation of new genetic diversity through breeding. When working with NUCs it is not always possible or necessary to set up a breeding program. According to the local bio-physical and socio-economic requirements in the target value chain, a genetic resources dynamic management or cultivar testing approach might be as well a suitable approach. In other cases, given the impossibility to introduce a certain NUC in a region or the foreseen market opportunity, a breeding program can also be suited. In agricultural regions where a crop has been grown for generations, a primary focus is suggested towards the improvement of existing landraces rather than their replacement with new cultivars, unless there are challenges regarding current farming and food requirements that can only be overcome through breeding. This approach has environmental and economic advantages as it allows to conserve agrobiodiversity and it is less costly compared with cultivar development. Yet, feasibility in terms of integrating the landraces in nowadays agronomic management practices or restoring traditional ones shall be carefully considered according to the cropping system and value chain of reference. One example of such a tradeoff could be the choice between cultivating an indeterminate common bean variety, whose harvest cannot be mechanised, and developing a new bushy variety from the same genetic base. The recommendation to prioritise the use of landraces is founded on the recognition that they are dynamic populations that have evolved and continue to adapt to their specific local conditions through both natural and human selection. Consequently, they possess a valuable genetic heterogeneity that provides resilience and yet a degree of inherent adaptation to their traditional management systems and uses. The central principle here is to preserve their agricultural significance by using innovative (e.g. participatory) selection and/or breeding approaches to shape their ongoing evolution. By integrating traditional knowledge and on-farm selection for current agriculture practices needs, this approach ensures that improvements are relevant and beneficial within the crop's existing cultural and ecological context. Page 16/35 Info BOX 1 The role of technical and social innovation in NUCs breeding In general, innovation in plant breeding involves the continuous development of new methods, tools and strategies to enhance the development of plant varieties. Although the term often calls to mind images of cutting-edge technical innovation such as molecular biology tools, innovation in this field is not confined to scientific breakthroughs. For sure, technical innovations have mirrored advances in scientific knowledge, especially in the 20th century when deeper insights into plant physiology, genetics and molecular biology enabled the development of new crop varieties with knowledge of the target crop that was previously unimaginable. These scientific advances surely increase precision and accelerate genetic gain. However, seeds are not just another agricultural input. They are also a cultural and ecological commons, carrying histories, traditions, and socio-ecological relationships. This means that plant breeding must also embrace social innovation—new ways of organizing, collaborating, and sharing knowledge that ensure breeding serves diverse communities and ecosystems. Social Innovation in plant breeding encompasses participatory plant breeding (PPB), commons-based seed networks, collaborative governance models, open-source seed initiatives and inclusive frameworks that empower local value chain actors. Participatory approaches involve farmers, scientists, breeders and other value chain stakeholders as equal partners in setting breeding goals, evaluations, selections, and seed management, building trust, local ownership, and adaptive capacity. Approaches such as participatory plant breeding are crucial, especially for crops that are underutilized and overlooked by industrial breeding systems. These social innovations ensure that breeding responds to diverse local needs, empowers communities, and fosters long-term sustainability by embedding breeding within broader social and agroecological systems. Innovation in plant breeding, is most effective when it integrates the technical and social dimensions (Figure 9). While technical innovation provides powerful breeding tools and a deeper understanding of genetics and phenotypes, social innovation ensures that these tools are embedded within equitable, contextsensitive, farmer-centred systems. 2.6. More breeding for complex cropping systems When trying to include minor crops into diversified farming systems, we also encounter the limitation of little technical research applied to these more complex systems. For example, intercropping of grain legumes with cereals can promote an agroecology-based intensification of low-input systems, along with other advantages (e.g., greater cereal protein content; reduction of costly and/or environmentunfriendly inputs). On the other hand, obtaining a balanced legume-cereal mixture is frequently hindered, for the legume species components, by insufficient competitive ability of the legume against the cereal. As the performance of genotypes grown in pure stand as monocrops is not necessarily a good indicator for their performance grown in intercrops, key questions for breeders are for which traits one can select in monocrop environment and for what others the selection directly in intercropping setting is necessary. At the moment the selection of varieties specifically adapted to intercropping remains a major practical challenge to its widespread deployment and requires effort and resources (Rubiales et al., 2023). Page 17/35 DIVINFOOD values the use of minor crops in diversified farming systems. To promote agroecological management practices, DIVINFOOD works on selecting and testing the minor crops in the context of intercropping in several of the LLs. APPROACHES 2.7. Breeding for Diversity When developing cultivars for agroecological and organic systems, alternative approaches might be of interest compared to breeding for conventional farming systems. For example, heterogeneous materials, that are intra-specific genetically diverse cultivars offer greater adaptability and resilience. Breeding for Diversity (Chable et al., 2020), which prioritize genetic diversity and decentralized adaptation are particularly well-suited for organic and agroecological farming due to their ability to enhance resilience against biotic stresses (such as pests and diseases) and abiotic stresses (such as drought or poor soil conditions). By reducing reliance on chemical inputs, high genetic diversity cultivars (such as evolutionary populations, open pollinated varieties, composite cross populations, organic heterogenous materials, farmers selections) align with organic farming principles. Additionally, high genetic diversity cultivars contribute to ecosystem services. Their dynamic adaptability allows them to evolve in response to local conditions, making them ideal for low-input systems like organic farms. The EU Organic regulation (Regulation (EU) 2018/848) recognition of "Organic Heterogeneous Material" (OHM) further supports the use of high genetic diversity cultivars in Europe. For minor crops, breeding for high genetic diversity cultivars often offers a good cost-benefit balance. In fact, this method helps to preserve genetic diversity in minor species, while also creating niche market opportunities. Breeding of genetically diverse cultivars should be prioritised (population breeding, organic heterogenous material breeding). The decision of developing and heterogeneous materials rather than homogeneous varieties, should be made in consultation with all relevant actors of the target value chain to minimize the trade-off of the different options. Generally heterogenous materials have the highest potential in local value chains, for quality products and cultivation in highly diversified farming systems and in marginal conditions. DIVINFOOD values the use of novel approaches to breeding. To promote agrobiodiversity and tailored markets, DIVINFOOD works on developing heterogeneous material for pea, einkorn, emmer and poulard wheat (in Italy, France and Hungary, Appendix 3). 2.8. Align breeding with the ethical values of involved stakeholders The development of NUCs is not solely a scientific problem to be solved with technology; it is a moral and societal issue regarding agrobiodiversity conservation and food diversity and inherent cultural diversity. Breeding programs that are not transparent and are not aligned with the values of their Page 18/35 stakeholders—such as a respect for traditional knowledge or a commitment to food sovereignty—are likely to face resistance and fail to achieve their goals. This principle moves beyond technical feasibility to address the ethical dimensions of plant breeding, encompassing a complex set of principles, including environmental stewardship, social equity and respect for living organisms. The selection of breeding methods and approaches for NUCs must be guided by the ethical and normative attitudes of all involved stakeholders. A multi-actor consultation by the DIVINFOOD project will summarize a vision for future NUC seed systems in Europe is a concrete application of this principle. This approach recognizes that the success of a breeding program is fundamentally tied to the trust and buy-in of the communities it serves, making transparency and accountability non-negotiable components of the process. 2.9. Promoting Access to Seeds A core tenet of NUC development is the prioritization of farmer access to seeds, including the crucial practice of on-farm seed saving. The facilitation of community seed banks (CSBs) and/or SME-based local seed production is a crucial strategy for building a resilient seed sector for NUCs. Community seed banks are far more than simple conservation tools; they are foundational economic and social institutions. They provide a vital function by offering farmers, particularly smallholders, access to locally adapted varieties that may be unavailable or too costly through formal systems. Working with NUCs can be also a business occasion for small scale professional seed producers, combining seed access with rural development and job creation. In the LL Cer-Occ a simplified version of the FAO’standard Material Transfer Agreement (sMTA)2 was developed for allowing a more easy and understandable exchange of seeds among farmers. OMKI is involved in activities related to seed quality in organic farming in order to develop capacity bulding on different aspects of seed production Figure 10. White lupin seeds (Photo: Mariateresa Lazzaro) 2The Standard Material Transfer Agreement (SMTA) is a standard contract that sets out the terms and conditions ensuring that the relevant provisions of the International Treaty are followed when transferring plant genetic material that is included in the Multilateral System. https://www.fao.org/plant-treaty/areas-of-work/the-multilateral-system/smta/en/ Page 19/35 COLLABORATIONS 2.10. Multi-Actor involvement A core recommendation based on the DIVINFOOD LLs experience for the success of NUCs seed sector advancement is to work with an inclusive participatory approach open to all value chains’ actors (Info BOX 2). This approach provides a holistic picture of desirable traits along the whole value chain, while contributing to evaluate cultivars and co-develop protocols and recipes for the production, processing and consumption of NUCs. In DIVINFOOD, stakeholders collaborate within LLs with the ultimate aim to set up territorial networks for biodiversity governance and management. A diverse set of actors is included, such as policymakers, farmers, citizens, consumers, food processors, gastronomists, retailers and other professionals of the agri-food system. In time DIVINFOOD aims to support the transformation of LLs into new territorial multi-actor networks in charge of including local stakeholders’ needs, co-manage breeding, and organise seed multiplication. The implementation practices of the participatory and multi-actor approach are very diversified, as each Living Lab acts in different local and regional environmental, agricultural, political and socio-cultural circumstances. Because of this, we developed a framework for summarising these experiences and showcasing pathways for future multi-actor activities working on NUC breeding, cultivation and use (Info BOX 3). An important point of reflection for the networks involved in developing the seed sector for underutilised crops is which stakeholder groups are involved. Awareness on this aspect is necessary for goals setting and appropriate network facilitation and governance management. There can be (Figure 5., lower axis) more research-centered approaches, such as the cases of research organizations reaching out to different actors and initiate and conduct participatory activities to support the NUC promotion process; and also the other way around, more community-based approaches, i.e. actors are initially motivated and trigger the collaboration, e.g. a farmer collective that starts crosses and selections to develop own peasant seeds without any prior input from research organizations. For example, Leg-It-Switz is on the right side at the breeding axis, as it has established formalized breeding programs for white lupin and pea, building on a broad range of accessed and generated genetic diversity, potentially holding multiple beneficial traits for cultivation, processing and consumption and allowing for selections of the best suitable materials. The core breeding work is performed in a professionalized setting and supported by the scientific expertise in molecular breeding and bioinformatics by the research institute to search for complex trait constellations which would make the utilization of the NUC more attractive to the agri-food sector, e.g. anthracnose tolerance combined with grain sweetness in the case of white lupins. The level of multi-actor involvement in the breeding work is rather low; however, networking to connect to farmers, processors and consumers as well as awareness raising activities for citizens have been organized to trigger market development and demand. Similarly, in Italy, farmer evaluation is considered for cultivar selection at the latest stages of cultivar development. A specular example, the Living Lab Bean-Lyon works on a single cultivar, the hairloom bean ‘Haricot viande’, so it is on the left side of the breeding axis. Here, the approach is not breeding by improving traits and selecting out of a range of genotypes but building up and adapting the agri-food system around a specific genotype with a local tradition of cultivation. To fulfil this, several actors from farmers to processors and gastronomists to citizens, consumers and policymakers are involved to co-develop a way to cultivate Page 20/35 and consume this specific genotype. By bringing a forgotten cultivar back to utilization, agrobiodiversity is enhanced on the farm, crop and nutrition level by diversifying crop rotations and increasing local bean production. The specific cultivar is allowed to be maintained not only ex-situ but in-situ, while new genetic diversity is not generated Info BOX 2. Participatory Plant Breeding & Multi-Actor Involvement: a holistic approach to enhance agrobiodiversity Participatory Plant Breeding (PPB) is an approach to develop genetic material informed by the perceptions and experiences of end-users. The main advantages are the insights into trait importance and genotype evaluation in the real conditions of cultivation and use, which is connected to direct adoption prospects (Ceccarelli & Grando, 2007; Chiffoleau & Desclaux, 2006). Many successful PPB initiatives were implemented in the Global South (Song et al., 2023), where resource constrains make it a viable alternative to expensive and not necessarily most suited standard breeding programs. In Europe, PPB is increasingly emerging with several initiatives in place (Colley et al., 2021). PPB and is particularly interesting for the NUCs as it offers a frugal and locally based option. Participation activities tend to focus on farmer involvement, with experiences and attempts to engage other stakeholders often including much less structured approaches and less direct influence on the breeding outcome. However, as urging challenges such as climate change, (agro)biodiversity loss, land degradation and the environmental footprint of agricultural and food production call for a systematic shift in the whole agrifood system towards diversity, resilience and input reduction, it becomes obvious that the development of future genetic material and associated agricultural practices demands a holistic approach involving all concerned parties of the agri-food system (Colley et al., 2022). After a century of scientific and economic specialization, segmentation and optimization, a perspective of re-learning and re-connecting to ecologically valuable agroecological practices such as the cultivation and consumption of neglected and underutilised crops (NUCs) is taken, supported by the pressure of the organic and agroecological movement and environmental and social activists. In the light of these developments, it is important to extend the concept of Participatory Plant Breeding to the methods of Multi-Actor Value Chain Creation, with the objective to fully involve all stakeholders in the value chain, including farmers, food producers, chefs, retailers, consumers, citizens and even policy makers in the development (breeding), cultivation (farming) and use (food system) of locally adapted genetic material of NUCs. While it is easy to understand the need for this multi-actor approach, the development of the concept and its implementation are emerging together with the attempts to combine PPB and suitable value chain co-creation for the genetic material outcome of participatory breeding and selection programs. Page 21/35 Figure 11. DIVINFOOD team members making an experience with grey pea (left), einkorn and emmer (right) participatory plot evaluation at annual meeting 2024 and 2025 respectively (Photo: DIVINFOOD 2.11. Goals setting in Multi-Actor Breeding Given the interest to work in a multi-actor setting, a further domain of reflection (especially for the multi-actor network facilitators) is about which GOALS can be reached given the starting point of a certain LL or value chain. We developed a framework for the goals in multi-actor breeding (Figure 8) to improve clarity about different domains and targets, which can be used as guideline on the opportunities and limitations of participatory activities. This framework can support multi-actor networks in designing participatory activities. This is considered important, because participation needs dedicated preparation, facilitation and evaluation, which means a high level of human resource effort. As most programs on NUCs work under limited resources, informed decisions about who to involve when and overall well-planned activities can maximize the impact. The framework also demonstrates, based on the DIVINFOOD LLs, that many actions are not taken to help the selection in any given panel of cultivars (evaluation/selection), which can be understood as strictly participatory breeding in the narrow sense, but rather to establish and improve the conditions for the NUC to come into cultivation and consumption (capacity building), appreciated by the society, politics and the agri-food stakeholders (awareness-raising) in the context of multi-actor value chain creation. We grouped the GOALS in multi-actor initiatives for NUCs in: awareness raising, capacity building and evaluation/selection from the closest to value chain creation to the closest to participatory cultivar development (Figure 12). Of course, the different goals are inter-connected and all together support the inclusion of NUCs from “plate to seed”, yet topics (target of awareness raising activities), domains (of capacity building activities) and traits (for evaluation/selection) are different across these three goals. We suggest to use this scheme for making clear as a network / LL which activity is conducted for which goal as first step to optimise human and economic resources. The Awareness-raising goal focuses on setting up the broader context of importance and value to maintain and promote NUCs by stimulating consciousness for the overarching topics such as agrobiodiversity and socioeconomics, ethics and justice in the agri-food system, including the seed system (seed legislation, seed market, management of genetic resources, breeding, multiplication Page 22/35 and seed access and distribution). The specific local case of a NUC is connected to broader questions on the relationship between society and nature. This includes aspects such as exploring how resilience towards climate change and intraand inter genetic diversity are entangled, uncovering current political and economic structures and practices hindering the mainstreaming of agrobiodiversity, explaining critical aspects of technology-focused breeding methods and acknowledging which diverse goods and services are delivered by diversified food systems. For example, these goods can be the promotion of human health through diverse plant-based diets and valuable nutrients and tastes of NUCs; cultural, aesthetic and heritage values by (re)learning farming and processing practices, value forgotten handcraft skills and culinary traditions and rediversify agriculture on the landscape level; and ecosystem services, e.g. beneficial traits of NUCs like deep rooting and nitrogen fixation for soil health or increasing cultivation of legumes to the benefit of pollinators. By taking this wider picture into account, participants of all kinds of actor groups are addressed in their role as citizens (and eventually policymakers) in their respective political system. These activities empower to take more informed decisions in their specific settings (genetic resource management, farming, food manufactures, shops, kitchens) and invite to join political forces and build broad alliances for enhancing agrobiodiversity. This groundwork of understanding and motivation can be seen as a perquisite to work on the goals of capacity building and selection/evaluation, which are more technical and specific. Importantly, this is not a one-way process, as the participants might bring additional critical reflections and insights from their own backgrounds into the discussion. In the context of awareness-raising another approach can be on actor-specific aspects of NUC production by introducing the challenges but also opportunities they pose. Farmers can be made aware of additional crops they have not considered so far. Their prospects can be discussed, such as widening crop rotations, reducing agricultural inputs and making better use of marginal lands by using low-intensive crops such as NUCs, as well as diversity, health and heritage values as marketing factor. Also, their challenges should be explained, such as seed availability, lower yield levels, lodging tendencies or technical difficulties in harvesting and sorting/cleaning. Awareness on the possibility to extend the product range, develop new attractive and healthy recipes and diversity, health and heritage values as marketing factor will be informative to processors and gastronomists, as well as discussing challenges such as varying raw material availability, need of specialized infrastructure and processes and the effort to explain new products and eventually increased price levels to consumers. On the consumer side, awareness about challenges for farmers and processors increase understanding for premium price setting and should also emphasize the responsibility of pro-active, critical consumers who shape their own local food supply environment, e.g. by suggesting portfolio changes to retailers or take account of direct-marketing structures. The capacity building goal goes beyond awareness raising, towards the development of practical knowledge and infrastructures to make NUC cultivation and consumption a reality. The working modus in this domain is therefore participatory, practice-oriented research to try out, document and disseminate the best ways to cultivate and process NUCs and to facilitate the exchange between all actors along the value chain and integrate established and new knowledge for best praxis. The domain breeding refers to encouraging farmers to make their own on-farm selections for siteadapted cultivars. Co-research in the domain agronomy includes on-farm trials for developing suitable cultivation protocols, e.g. the search for a proper companion crop for intercropping systems or the right sowing density, weed management options or the point of harvest and settings for the threshing machine. It also means collecting feedback by farmers on the practicability to cultivate the respective NUC in their specific setting, and what needs to be improved in their view via Page 23/35 improved/more suitable cultivars or cultivation system modifications. For processing, new products and recipes can be introduced to or co-developed with food processors and, together with food retailers, they can estimate the technical and economic feasibility and marketability prospects and strategies. Consumers contribute with their feedback on products and can also be accessed to be instructed on or co-develop protocols and recipes for home cooking; triggering market demand by consumers informed about how to process the product into a tasty dish. The Evaluation/Selection goal has the strongest focus on the seed system. It refers to multi-actor involvement in the sense of participatory breeding, meaning the integration of views of different actors into trait definition and selection in a cultivar panel of any kind (e.g. landraces, populations, breeding lines). Improved decisions on trait importance informed by relevant actors and consideration of additional traits will benefit the selection process to find suitable cultivars. It also opens the interesting research area of comparing the effect of selections by different actors (e.g. between farmers and professional breeders’ decisions). In this realm, actors are invited to evaluate the intra-genetic diversity, e.g. a tasting session with consumers to select for the best taste, texture and appearance out of a cultivar panel, or to ask farmers to choose their favourite plots in a cultivar panel or their favourite single plants in segregating material, and/or to describe and rate the plots/single plants for criteria relevant to them. Info BOX 3 Methodology: summarize the experience of the DIVINFOOD Living Labs The implementation practices of the multi-actor participatory approach are diversified in DIVINFOOD, as each Living Lab acts in different local and regional environmental, agricultural, political and sociocultural circumstances. We used the opportunity of these diverse examples to develop a framework for reflection on these experiences and showcasing pathways for future multi-actor activities in these and other initiatives working on NUC development and promotion (Figure 5, Figure 12, Figure 14). Semi-structured interviews with DIVINFOOD Living Labs coordinators have been conducted in 2024, to get an understanding of local and crop-specific tasks and challenges and the structure of the Living Lab and concerned stakeholder networks and its connection with the seed system. In these interviews with the LL coordinator, the concrete activities with multi-actor involvement were investigated. The results have been structured into a summary per type of events, actor groups involved, methods used, implementation status, expectations and output, supporting factors and challenges. Based on this overview, we developed the guidelines related to (1) material type, (2) actor involvement, (3) goals in multi-actor involvement for the different actors and (4) specific goals when involving members of the general public in their role as citizens vs consumers. These frameworks, illustrated as infographics, showcase the overarching topics in multi-actor involvement which were arising from all interviews and illustrating common themes and pathways despite a highly local-specific context in each LL. Page 24/35 Figure 12. Goals in Multi-Actor Breeding (source: authors) DIVINFOOD – H2020 n°101000383 f Page 25/35 2.12. Involvement of consumers and citizens As citizens/consumers are often involved in value chain creation but more rarely in seed and breeding sector development, based on the DIVINFOOD experiences we summarised the involvement framework for these specific actor-groups (Figure 14). Citizens and Consumers are often seen as the ‘general public’ actor group but have quite distinct characteristics and therefore different implications in activity design, when taking a theoretical and planning look at their involvement. Of course, a participant can have the role of both a consumer and citizen, and on most occasions, this will be the case. Anyway, for the activity organization and the estimation of the expected output, it is helpful to be clear about the different aims and impact pathways depending on which role the participant is expected to take. Of course, any event like an open field or food production day could include several of these pathways, so that both roles are reached on the same day. In the awareness raising pathway, people are addressed as citizens, which is a role every invited person to any event has, next to more specific roles such as farmer, processor. The goal is to create understanding about what could be the value of NUCs for a diversified food system. This can happen on the species level (Figure 14, with citizens, left) by focusing on one NUC or several NUCS, eventually compared to cash crops, experienced on-field and/or through product tastings. So, the activity has a focus on sensory, direct impressions. The usual approach will be inputorientated through a guided tour; however, it is possible to include interactive elements. For example, participants could be invited to estimate the yield levels of a NUC compared to a common cash crop or describe their overall impression of the NUC. On the methods level (Figure 14, middle), with the starting point of NUCs values and challenges, different types of cultivars, breeding approaches and implications of the choice of cultivar development strategies and how this is connected to seed policies and the seed market can be discussed. This pathway has a more instructive and cognitive character compared to the left and right pathways, as it deals with specific sector knowledge which is not known to the public in most cases. It can be supplemented by a group work part to deepen the level of engaging with the topic. For example, participants could be asked to brainstorm what the Distinctiveness Uniformity and Stability (DUS) system for variety registration means for small-scale seed actors, collect drivers for the decline of agrobiodiversity or visualize strategies for enhancing diverse agri-food systems. Group exchange and feedback from the participants will be a source of valuable information about how they perceive these issues, which could be used for further awareness-raising campaigns or feed into the capacity building domain, e.g. to inform strategies for marketing. At the varietal level (Figure 14, right), citizens are invited to experience the intra-genetic variance in a NUC cultivar panel and gain insights about different possibilities and ranges of trait expression and approaches for evaluation and selection in this cultivar panel. This can be done on-field and/or with a panel tasting session. Like at the species level, it is a sensory-orientated approach, usually based on an input-focused guided tour and supplemented with exercises amplifying the understanding. For example, participants could evaluate certain easily observable traits such as earliness, vigor, plant health or lodging susceptibility and experience first hand typical trade-offs in breeding selection, e.g. if the healthiest cultivar is also the one most prone to lodging or if the earlier cultivars, which might be favorable for farmers wishing for an early harvest, have low vigor and yield potential. In the capacity building pathway participants are addressed in their role as consumers (Figure 14, Goal 2). The focus is on triggering market demand by codeveloping, improving and sharing advise on concrete NUC-based products and recipes which will be suitable for home or gastronomy consumption. There are two pathways in engaging the consumers for capacity- DIVINFOOD – H2020 n°101000383 f Page 32/35 Appendix 2 Examples of molecular breeding activities in DIVINFOOD Living Labs DIVINFOOD innovative techniques Crop Living Lab (Country) Description Organisation Molecular breeding of grasspea Grass pea (Lathyrus sativus) GPea-Port (Portugal) • Collection of 182 grass pea accessions representative of the species' worldwide cultivated germplasm diversity, being composed by traditional landraces, breeding lines, and varieties. • A total of 16 agronomic traits were scored according to the “Descriptors for Lathyrus spp.” (IPGRI 2000). • A panel of 5,651 high-quality Single Nucleotide Polymorphism (SNP) markers was used to test for SNP-trait associations • To our knowledge this is the first grass pea Genome Wide Association Study (GWAS) reporting putative mechanisms of agronomic adaptation and pest resistance genetic control, paving the way to the development of molecular tools to assist precision breeding toward achieving multi-stress resistance grass pea varieties. >> Results to be published Universidade NOVA de Lisboa (UNL) Molecular breeding of lupins White lupin (Lupinus albus) Leg-ItSwitz (Italy) • Research 1 : Improved adaptation to autumn sowing in organic farms of Northern Italy. 150 inbred lines are being tested under autumn sowing in Northern Italy for grain yield, with a farmer assessment day (organized by FIRAB). Genomic selection models will concurrently be developed for grain yield (exploiting project data as well as earlier phenotyping data, using already available molecular data). Concurrently, 16 top-performing lines selected phenotypically for yield and farmers’ acceptability are being tested in an organic farm. >> Results to be published • Research 2: Improved adaptation to moderately calcareous soils. The aim of this work is to validate a genomic selection model for lime tolerance established from previous projects, applying it to selection within 450 independent lines whose genotyping data are already available. CREA is performing this proof-of-concept experiment in large managed environments Council for Agricultural Research and Agricultural Economy Analysis (CREA-ZA) DIVINFOOD – H2020 n°101000383 f Page 33/35 with contrasting soil types, including material that is predictively besttolerant, mid-tolerant. and bottom-tolerant to soil lime. >> Results to be published Molecular breeding of lupins White lupin (Lupinus albus) Leg-ItSwitz (Switzrland) • Bulked Segregant Analysis of an F2 population, segregating for two different low alkaloid conferring loci • Identification of a novel QTL on chromosome 5, • Novel source of sweetness apart from pauper and PCR marker developed • 50 stacked allele recombinants were identified that exhibit an exceptionally low total alkaloid content (22.8 ± 10.4 ppm), even when compared to genotypes known to carry the allele associated to the most drastic reduction in QAs, pauper (171.7 ± 18.5 ppm). >> Publication: https://doi.org/10.1186/s12870-025-06951-7 Research Institute of Organic Agriculture (FiBL) DIVINFOOD – H2020 n°101000383 f Page 34/35 Appendix 3 Examples of population breeding in DIVINFOOD Living Labs DIVINFOOD innovative techniques Crop Living Lab (Country) Description Organisation Pea population breeding in Italy Pea ( Pisum sativum ) Leg-ItSwitz (Italy) • This work aims to assess the adaptation and agronomic value of evolutionary populations relative to pure lines under on-farm conditions in Northern Italy. In one organic farm, we will compare one population that is being notified as Organic Heterogeneous Material (registered as ‘Ellen’) vs. the best pure line selected from the same genetic base for organic farms of Northern Italy (registered as ‘Pantera Rosa’) vs. the pea cultivar currently grown. This experiment will be carried out in the season 2025-26. >> Results to be published One pea population notified as Organic Heterogeneous Material (‘Ellen’). Council for Agricultural Research and Agricultural Economy Analysis (CREAZA) Emmer population breeding Emmer ( Triticum dicoccum) Cer-Hung (Hungary) • Heterogeneous populations of durum wheat were used as mothers in crossings with emmer landraces to develop hulless, high value, new emmer cultivar candidates. • Consecutive spike selection steps and bulk propagation - in the case of heterogeneous materials - resulted in lines and heterogeneous populations that exhibit various kinds of combinations of the parental traits. • Most promising materials have been tested from F4-F5 generations on farm with the participation of farmers so that evolutionary adaptation can take place, as well as to provide an opportunity for farmer selection from them. Hungarian Research Institute of Organic Agriculture (ÖMKi) Einkorn and poulard wheat population breeding in France Einkorn (Triticum monococcum) Rivet wheat (Triticum turgidum) Cer-Occ (France) • Einkorn and rivet wheat population breeding is ongoing in South of France • Concerning einkorn: the starting point of this initiative was the mobilization of 103 einkorn accessions from the Centre de Ressources Biologiques (CRB) at INRAE Clermont-Ferrand, a key repository for cereal genetic diversity in France. These accessions INRAe & École d'ingénieurs de Purpan DIVINFOOD – H2020 n°101000383 f Page 35/35 represented a wide genetic base, including traditional landraces and lines collected from different regions of Europe and the Mediterranean. • Concerning rivet wheat, the initiative began with the collection of seeds from CRB, from different seedhouses, from farmers, materials at EI Purpan. This collection represented a wide genetic diversity, from different regions of Europe.