H2020 CHIC Project: Chicory as a multipurpose crop for dietary fiber and medicinal terpenes
- Type Project
- Status Filled
- Execution 2018 -2022
- Assigned Budget 6.999.596,25 €
- Scope Europeo
- Main source of financing H2020
- Project website Proyecto CHIC
Chicory is a relatively understudied crop, so gene editing protocols had to be developed virtually from scratch. Variants of CRISPR methods were developed that differ in the extent and manner in which DNA is used to deliver the CRISPR system that generates the genome edits. These methods were systematically evaluated for efficiency and safety, with potential off-target editing being studied in each case. No off-target mutations caused by the CRISPR system were detected in any of the methods used or in any of the lines tested. When DNA was used to deliver the CRISPR system through stable transformation, different edits were observed within the same plant in some plants. Apparently, the stably integrated CRISPR complex continues to make edits at the remaining target sites as the plant develops. When DNA was delivered transiently with the CRISPR system, in addition to the intended edits, some plants also found fragments of CRISPR DNA unintentionally integrated into the genome.
In both cases, these plants can be recognized and eliminated. When the CRISPR system was delivered as a protein complex rather than through DNA, modifications only occurred at the intended target sites. This makes this method highly attractive for commercial applications. The methods were implemented to generate a multitude of chicory varieties: varieties with higher quality healthy inulin, as its degradation in autumn is prevented, as well as varieties with reduced bitter terpene content, greatly facilitating inulin extraction. Furthermore, varieties that accumulate specific terpenes with potential medicinal value were created. The latter was guided by bioassays that identified chicory root terpenes with highly interesting anti-inflammatory and antimicrobial activity. Finally, chicory's self-incompatibility was disrupted, simplifying crossbreeding and allowing for more efficient breeding. Through these experiments, scientific knowledge was generated about the biology of chicory and, in particular, about its bioactive compounds, their biosynthesis and storage.
A Stakeholder Advisory Group, comprising representatives from industry, academia, agriculture, and end-users, interacted with the CHIC consortium throughout the project period. Furthermore, consultations with a broader spectrum of stakeholders spanning the entire agricultural and food value chain, as well as with policymakers and regulatory agencies, helped clarify the factors that hinder and facilitate innovation in gene-edited plants in general and root chicory in particular. Scenarios differing in aspects such as whether CRISPR-edited chicory is regulated as a GMO or not, and what types of products are isolated from it, were evaluated based on their socioeconomic and environmental impacts along the entire value chain. These results demonstrate that inulin and/or terpene production based on the new chicory variants generates more employment, higher added value, and reduces greenhouse gas emissions and primary energy demand compared to the current process. Particular emphasis was placed on communication about the CHIC project and its objectives.
A website was created with access to brochures, newsletters, and explanatory videos, as well as social media channels. Artists visited laboratories and created artworks that enabled alternative interactions with the general public. An educational game (MyChicFarm) was developed, and CHIC partners visited schools and organized roundtable discussions specifically aimed at young people. CHIC also monitored global regulations on genome editing. In the EU, the Court of Justice of the European Union ruled that all gene-edited plants are regulated as GMOs. The regulatory landscape is currently changing rapidly, with significant differences across continents and countries. The project clearly demonstrates that promising food applications will only be pursued by EU companies if the regulatory status of certain gene-edited plants changes.
CHIC, the Chicory Innovation Consortium, was a highly multidisciplinary project whose objective was to develop genome editing (CRISPR) to transform chicory into a crop capable of extracting multiple products beneficial to human health. Simultaneously, we engaged with various stakeholders to gather their feedback on the innovation pathways developed.
Chicory is cultivated on a relatively small area, mainly in the southern Netherlands, Belgium, and northern France. The dietary fiber inulin is extracted from its roots, which is used as an ingredient in products such as yogurts and bars to boost gut health and serve as a non-caloric sweetener. Chicory roots also accumulate terpenes, but these are discarded as waste from inulin extraction due to their bitter taste. Since terpenes from other plants have very interesting health properties, the potential of genome editing to turn chicory into a multipurpose crop from which improved inulin and beneficial terpenes can be extracted was investigated. Developing new chicory varieties through conventional plant breeding can easily take one to two decades.
Furthermore, chicory is self-incompatible, making crossbreeding difficult in plant breeding programs. Therefore, genome editing can make a difference. In public debates, genome editing often generates high expectations as well as strong concerns. Therefore, CHIC aimed to engage a wide range of key stakeholders to raise awareness and discuss issues related to both the chicory varieties developed in the project and genome editing in general. CHIC evaluated various genome editing methods for efficiency and safety and investigated potential commercial use scenarios for these edited chicory lines, considering their environmental, regulatory, socioeconomic, and overall societal impact. CHIC strived to communicate openly by implementing innovative communication methods. For example, artists have familiarized themselves with genome editing techniques and expressed their views and opinions in artwork to stimulate broader public debate.
CHIC is the Chicory Innovation Consortium.
Its objective is:
- Implement New Plant Breeding Techniques (NPBT) in chicory to establish it as a multipurpose crop for the production of health-related products with clear benefits for consumers.
Develop co-innovation pathways with stakeholders for innovative technologies, such as NPBTs.
CHIC will develop four different NPBTs. They will be used to direct bioprocesses in chicory and leverage its underexplored potential to produce immunomodulatory prebiotics and medicinal terpenes.
The conceptually diverse NPBTs will be assessed with respect to their technological potential, risks, regulatory framework, and socioeconomic impacts. This will be done in close consultation with a Stakeholder Advisory Group (SAG) composed of relevant stakeholders from industry and society. Ongoing project activities and results will be discussed with stakeholders and communicated to the interested public using innovative methods, including cultural communication and linking art and science.
In this context, CHIC will develop two business cases in different application areas: inulin as a healthy food ingredient and terpenes as medicinal compounds. This endeavor requires a highly interdisciplinary approach with expertise in molecular sciences, economics, arts, social sciences and humanities, and legislation. The partnership includes three SMEs and one chicory end-user, and international collaboration is established through a research institute in New Zealand. The SAG plays a crucial consulting role in all phases and activities of the project. Through this co-innovation approach, our aim is to contribute to leadership in responsible research innovation and promote a better understanding of plant biotechnology. Chicory will be promoted as a robust multipurpose crop, tolerant of harsh environmental conditions from which bioactive compounds can be extracted, contributing to sustainable agriculture and a bio-based economy.
Currently, chicory is cultivated for its natural probiotics, but scientists are investigating the enormous variety of medicinal compounds that could be obtained from the plant. Humans have valued chicory varieties for centuries, enjoying them for their characteristic bitter flavor (e.g., endive) and also for their blue flowers. Chicory root is cultivated to obtain inulin, a natural probiotic that boosts the immune system. However, this plant has much greater potential.
Chicory naturally accumulates large amounts of terpenes, natural compounds that plants use to defend themselves against herbivores or attract pollinators. Terpenes also possess a wide range of medicinal properties, making them a promising source for new medicines. The EU-funded CHIC project investigated the potential of advanced genome editing techniques to help chicory reach its full potential. The CHIC consortium worked to improve the quality of chicory and adapt it to produce both healthier inulin and sufficient quantities of medicinal terpenes. “Although many terpenes from other plants have medicinal properties, the medicinal properties of chicory terpenes were largely unknown,” says Dirk Bosch, group leader of applied metabolic systems at Wageningen University & Research and coordinator of the CHIC project.
The CHIC project demonstrated that chicory can efficiently produce not only healthy dietary fibers, but also terpenes with medicinal uses. “It can likely also produce terpenes for other applications, such as cosmetics, natural plant protection products, and even for watchmaking for biopolymers,” explains Bosch. “A bio-based green factory,” he adds. Chicory meets CRISPR. Developing a new plant variety through conventional plant breeding can take decades. CHIC sought to develop new varieties in a shorter timeframe, using genome editing to transform chicory into a multipurpose crop capable of producing dietary inulin and bioactive terpenes. Through the project, the CHIC consortium evaluated genome editing techniques, such as different variations of CRISPR Cas, which works like molecular scissors to make small changes to an organism’s genome.
The project was based on three pillars: the development and implementation of CRISPR protocols for chicory; research on the safety and socioeconomic and environmental impact of the new varieties; and, finally, communication with stakeholders and the general public. Advancing Knowledge of Chicory Due to the limited project timeframe, the team was unable to cultivate enough genome-edited varieties on a large enough scale to allow for extensive biomedical experiments. However, analyses of the available material confirmed the accumulation of products with health-promoting properties, either already known or newly discovered through the project. "Preliminary experiments with bioactive plant compounds at the end of the project looked very promising," notes Bosch.
The CHIC project achieved several successful outcomes. The team discovered promising new medicinal bioactivities of chicory compounds and developed chicory varieties that produce healthier medicinal inulin and terpenes. The work also expanded our knowledge of chicory biology, uncovering, for example, how and in which tissues these compounds are produced. Public debates The idea of gene editing in crops is still controversial, so CHIC organized several public debates and workshops with artists to foster discussion and, together with project scientists, connect with the general public and schools. CHIC social scientists organized targeted debates and symposia with stakeholders, both locally and across Europe. During the COVID-19 pandemic, the team also hosted online events. “These interactive activities were truly inspiring, and in the debates, each of the different audiences expressed diverse reactions,” says Bosch.
The CHIC project has clearly demonstrated the contribution of genome editing to a niche crop like chicory. At the start of the project, relatively few genetic and breeding tools were available, and no genome editing methods existed for chicory. Five years later, multiple improved varieties have been developed with potential benefits for consumers (inulin and healthy terpenes), ingredient producers (more efficient processing and use of residues), farmers (agricultural diversification and harvesting flexibility), breeders (genome editing methods and self-compatible lines), the environment (reduced primary energy demand, reduced GHG emissions, and more efficient land use), and the economy (higher added value and job creation).
CHIC primarily focused on traits with health benefits for consumers. Using the knowledge base established by CHIC, other traits were also able to be introduced into chicory in relatively short timescales. More broadly, CHIC demonstrated that genome editing can be a powerful tool to help boost agricultural biodiversity in Europe by improving niche crops, which have low investment potential. In synergy with other breeding and cultivation methods, this is highly relevant for maintaining food security and improving sustainable production while addressing the challenges of climate change.
- STICHTING WAGENINGEN RESEARCH (WR)
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