Implementation of New Plant Breeding Techniques for dietary inulin

Inulin is a dietary fiber with health-promoting characteristics mainly targeted on gut health. Inulin is extracted from root chicory and used in many food products. The yield and the length of the inulin molecule determines the value and health-beneficial character of this compound. Using NPBT we want to increase both the yield and the length of inulin. The natural production of terpenes in chicory makes the inulin extraction method less efficient and more costly. The other goal of WP2 is to reduce the production of natural chicory terpenes.  

The work done during the first two years of the project CHIC shows that we could generate mutant chicory plants in which the genes encoding  the inulin break-down enzymes are blocked using genome editing techniques thanks to the small alteration the CRISPR tools made in the plant DNA. All mutated plants have been analyzed in great detail on DNA level using various methods.

Figure 1. First NPBT mutated chicory plants with knock-out of inulin degradation genes in greenhouse
Figure 2. First NPBT mutated chicory plants with knock-out of inulin degradation genes in greenhouse
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Socio-economic and environmental impacts on the whole value chain

In CHIC we evaluate socio-economic and environmental impacts on the whole value chain of the new developed chicory crops. Therefore a socio-economic impact assessment and an environmental assessment of NPBTs and the whole value chain will be performed. In addition to these quantitative assessments, a qualitative research on societal issues hindering or facilitating chicory innovation will be applied. 

The first steps included collecting and screening information on the CHIC value chain and the current chicory cultivation and inulin, therefore a literature and database research was made. Information on current chicory cultivation, inulin production, terpenes and NPBTs was screened and summarized. Followed by the identification of socio-economic and environmental indicators.  

Statistics on international trade of root chicory and inulin from the UN COMTRADE and statistics on land-use, chicory production and yield from the Food and Agricultural Organization of the United Nations (FAO) have been analyzed. Figure 1 below shows the gross production value of chicory roots (including intermediate uses like seeds and feed) in 1,000 international Dollars. The five biggest producers of chicory roots, according to the available data, are Belgium, France, Netherlands, Poland, and South Africa. However, the main producer of chicory roots by far is Belgium.  

To identify environmental and socio-economic impacts of the new CHIC process and the resulting products (Figure 2) an environmental assessment using the methodology of Life Cycle Assessment (LCA) and a socio-economic impact assessment using an input-output modelling will be applied. The whole value chain (e.g. breeding, cultivation, processing) will be included in the assessment.  

The LCA will give information on e.g. greenhouse gas emissions, primary energy demand, water consumption, land use of the new value chain developed in CHIC. Within the socio-economic assessment, the impact of different NPBTs on economic and social indicators will be quantified such as GDP, production volume, growth, competitiveness, and employment as well as the distribution of wealth and income between different sectors and regions within the EU and the global economy. The results will be used to lead the development within the project in a sustainable direction. 


Figure 1 Gross production value of chicory roots (Datasource: FAOSTAT)

Figure 2 Value chain of the CHIC process

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CHIC project: Breeding Chicory Roots for Health Products.

Don’t miss our new leaflet where we explain the main objective of CHIC project: To breed chicory roots to bring health products to the society!

Download the new leaflet.

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CHIC project at the science programme FUTURIS

On Monday, 14th October 2019, the science programme FUTURIS, from the Euronews channel, made a documentary about the CHIC project.

They made several interviews to some of our partners and they also explained the benefits of implementing new plant breeding techniques, in chicory breeding, to convert it into a multipurpose crop and to obtain high value products for consumers’ health benefits.  

On the other hand, our project coordinator, Dirk Bosch had a special interview in the Euronews Bonus Section. He provided detailed information about why we selected chicory as a promising and a strong crop to be converted into a multipurpose one.  

The documentary has been translated into 12 languages and we want to thank the FUTURIS team for guiding us during the filming process and for bringing such a nice result.

Enjoy the documentary!

Source:

https://www.euronews.com/2019/10/14/chicory-roots-will-boost-your-gut-health-say-scientists

https://www.euronews.com/2019/10/14/the-deep-rooted-truth-about-chicory-root

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CHIC at the European Researchers’ Night in Indjija (Serbia)

IBISS, partner in CHIC Project worked on organizing the presentation regarding the results of our CHIC Project, though team member Jovana Petrovic directly participated in this manifestation.

On September 27th 2019, Jovana Petrovic from University of Belgrade, Institute for Biological Research “Siniša Stanković, National Institute of Republic of Serbia presented knowledge gained so far on health-beneficial effects of Cichorium intybus and its possibilities to be used on every day basis.

People seemed very keen to get to know with facts how this plant that grows on meadows nearby, could exert useful effects as a tonic, coffee substitute and prebiotic.

Since this manifestation is dedicated to popularization of science making it interesting to children of school age, retired people and overall families with children, this Researchers’ Night was a unique opportunity for people to get familiar with new breakthroughs in research that they could directly benefit on.

The presentation of the Project took place at Indjija, the intersection city between Belgrade and Novi Sad.

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Development of four conceptually different New Plant Breeding Techniques

The objective of CHIC’s in this area is to develop four different methods for genome editing of chicory. These methods differ in their degree of ‘DNA invasiveness’: the extent to which CRISPR genes are introduced into the plant’s DNA. In the end all four methods lead to identical genetic outcomes.

CHIC methods  related to the presence of CRISPR DNA in the chicory plant’s DNA

These two methods are based on stable integration of CRISPR genes in the chicory genome, leading to so called transgenic  chicory plants. After the CRISPR genes have done their work, the genes are removed from the genome. In the 1st method the crossing and selection will be used to select offspring plants without the CRISPR genes and with the desired mutation. In method 2, specialized enzymes will be used to remove the CRISPR genes from the plant’s DNA, so making crosses is not needed.

During the 1st two years of the project, thanks to applying the 1st method we have obtained transgenic chicory plants. These plants are now being analyzed for functional edits in the chosen target genes: genes involved in the production of the enzyme germacrene A synthase. This enzyme is essential for the production of terpenes.

Methods related to the non-presence of CRISPR genes in the plant’s DNA

These two methods are based on administering CRISPR tools to chicory cells without incorporating CRISPR genes in the plant’s DNA.

In method 3 plasmid-DNA that harbours CRISPR genes, is introduced into individual chicory cells. Plasmid DNA is best known from bacteria: circular DNA molecules that encode important traits. After introduction into a chicory cell, the CRISPR DNA is expressed and the CRISPR tools are being assembled in the cell. The plasmid DNA is eventually degraded. 

In method 4 the CRISPR tools, protein and guiding RNA, are assembled outside the plant cell and then introduced into plant cells. This way there is no CRISPR DNA entering the plant cells.

For all methods we need to treat individual chicory cells (protoplast). The challenge then is to grow complete plants from these single cells. We have shown that this is indeed possible. We have also optimized techniques to introduce plasmid DNA and the CRISPR tools into chicory cells.

This has already resulted in plant cells in which the genes involved in the production of the enzyme germacrene A synthase are permanently disabled, thanks to the small alteration the CRISPR tools made in the plant DNA.

Optimizing the CRISPR tools

One of the CRISPR tools is a guide RNA (gRNA) molecule. The other is an enzyme called Cas9. The gRNA guides the Cas9 enzyme to the place in the plants’ DNA where a mutation is desired. We have designed gRNAs and have shown that they indeed guide the enzyme, enabling it to temporarily break the DNA at the correct place.

We have obtained plants with desired mutations in both chromosome sets (chicory is diploid) and in many or even all the gene copies chicory possesses. We are now testing the plants for the terpene content that may have some type of bioactivity.

Genome insight

The existing DNA screening techniques are not effective to screen plant material for small changes in the DNA in a high throughput manner. Therefore we are developing a new technique, based on techniques one of the partners developed for kiwi.

During 2019, the partners decided to jointly invest into a better assembled genome sequence of chicory, as the available genomic databases were of insufficient quality for our research.

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Great success of CHIC project in the exhibition held by Fondazione Edmund Mach in the 2019 European Researchers’ Night Edition

On Friday 27th of September, 2019 Fondazione Edmund Mach (partner in H2020 CHIC project) made an exhibition of CHIC project.

University and school students, retired people and families could see in vitro cultures and protoplasts through the microscope and do some tasting of our nice CHIC products.

 

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Plant Gene Edit­ing panel dis­cus­sion

On 11 September 2019 Michiel de Both (KeyGene) visited the Vikki Plant Science Centre of the University of Helsinki (Finland), to give a seminar as an invited speaker in their seminar series in plant science. The seminar covered genome editing technology in crop plant breeding, as well as the CHIC program, and was attended by ~80 people.

Because of the strong concerns in the Finnish plant science community about the 2018 Court of Justice ruling, the institute organized a panel discussion to debate issues of regulation, IP and science policy related to genome editing.

The panel consisted of a representative of the Finnish seed company Boreal Plant Breeding Ltd., of the Finnish Food Safety Authority Evira, of Kirsi-Marja Oksman-Caldentey of CHIC partner institute VTT, of Michiel de Both on behalf of KeyGene and of Alan Schulman, the current president of the European Plant Science Organization EPSO.

Journalists and politicians present in the audience took part in the discussions, which concluded with the realization that any change in the European regulation must be instigated by the EU member states, supported by informing stakeholders and the general public of the importance of plant breeding and innovation for the European economy.

 

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CHIC at the European Researchers’ Night 2019

During the night of September 27th the Museum of Science in Trento (MuSe) will be part of the European Researchers’ Night 2019, and it will host the staff from Fondazione Edmund Mach (FEM) to talk about their research topics.

The event is free and open to all the people, from kids to elderlies, starting from 6PM and lasting until midnight. It will consist both of short talks and demo/hands-on sessions, where the public will be involved with different science topics and invited to ask any question related to the research activity.

For CHIC project, scientists will talk about New Plant Breeding Technologies and Cas9 delivery inside plant cells, showing in vitro material (plants, protoplasts at the microscope) and explaining the challenges faced while doing Genome Editing in industrial crops.

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Fascination of Plants Day 2019 Success Stories

The biennial international Fascination of Plants Day (FoPD) is an event encouraging people from around the world become fascinated and enthusiastic about plants. Coordinated by the European Plant Science Organization (EPSO), it takes place globally on May 18, every uneven year since 2012.

In 2019, for it’s fifth edition, FoPD was a huge success gathering 862 events across 52 countries! Thanks to our 56 national coordinators, thousands of enthusiastic event organisers, partners, 5 national patronages and sponsors we have created real success stories. To know more about them, check our Fascination of Plants Day Success Stories 2019 publication.

In 2021, Fascination of Plants Day will be back for a new adventure. While waiting for its launch you can follow us on Twitter (@PlantDay18May), Instagram (@fascinationofplantsday), Facebook, Youtube and on our website: http://www.plantday18may.org

We welcome you to join us in preparing for Fascination of Plants Day 2021! Get organizing events in your country and keep tweeting about plants (#PlantsDay, #FoPD). We look forward to seeing you all in 2021!

Contact: Global Coordinators: Alexandra Barnoux, EPSO, BE; Trine Hvoslef-Eide, Norwegian University of Life Sciences, NO, Przemysław Wojtaszek, Adam Mickiewicz University, PL; Karin Metzlaff, EPSO, BE

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