KASP marker development for time of flowering and Ascochyta resistance in chickpea

The developed of a pair of NILs (Near Isogeneic Lines) for flowering time allowed to identify new targets associated this trait. In the main figure above, a pair of chickpea NILs is presented, at flowering time with early genotype showing flowers and late genotype showing no flowers. The gene CaELF3a, in chromosome 5 (Ca5), was identified as a prominent candidate, together wiht other homologs of flowering-related genes in Arabidopsis. This study unveil new insights into mechanisms governing flowering time in chickpea. It is being the basis for developing new KASPTM markers to accelerate marker-assisted breeding in chickpea programs.  

Ascochyta blight, caused by the fungal pathogen Ascochyta blight (AB), is a devastating biotic stress that poses a significant threat to chickpea cultivation, infecting all above-ground parts of the plant (Figure below). A region strongly associated with AB resistance in Ca4 has been detected in the framework of Task 4.3 – Genotyping collections and research populations, led by IFAPA. The aim is to develop KASP markers to select efficiently resistant genotypes.

Ascochyta blight lessions. Left to right in leaves, stems and pods.
The BELIS project has received funding from the Horizon Europe research and innovation programme under the Grant Agreement N°101081878.

Soybean activities within the southeast-European group

Research groups from CREA (Lodi, Italy), University of Zagreb, Faculty of Agriculture UNIZG FAZ (Croatia) and Institute of Field and Vegetable Crops Novi Sad IFVCNS (Serbia) are the partners within the WP4 Genetic effects and genotyping tools in legumes and WP5 Proof of Concept – Breeding of grain legumes of the BELIS project, with activities focused on soybean (Glycine max L.) adapted to southeast-European growing conditions. Main phenotyping trials for our work on developing the genomic selection models for selection of various traits (grain yield and quality, drought tolerance, etc.) will start in 2025, but in 2024 we are actively preparing genetic materials which will be included in trailing. At the experimental field of University of Zagreb, Faculty of Agriculture, a set of 115 soybean genotypes (cultivars and breeding lines) has been drilled in late April this year for the purpose of seed multiplication and screening of flowering and maturity time.

Drilling has been executed in almost perfect conditions and warm and rainy Spring allowed the good establishment of genotypes in the field. The same set of material has also been drilled at the experimental fields of the Institute of Field and Vegetable Crops Novi Sad as backup, while some additional genotypes are also being growing with our partners from CREA (Italy). We are hoping for the good growing conditions in the following months and, hopefully, for the successful harvest in the Autumn which will enable us to continue smoothly with further activities.

The BELIS project has received funding from the Horizon Europe research and innovation programme under the Grant Agreement N°101081878.

Unmanned Aerial Vehicle technology in red clover breeding programmes

An important goal of BELIS is the implementation of UAV (Unmanned Aerial Vehicle) or drone technology in practical breeding programmes.

BELIS foresees two approaches with UAV technology in red clover: (1) the development of a model to predict the amount of flower heads – an important predictor of seed yield – in field plots with red clover, and (2) the application of a model for biomass prediction in practical breeding programmes.

Up to now, ILVO shared a protocol for UAV data collection with the partner breeders (DLF, DSV, Agroscope and Barenbrug). In 2024, partner breeders will set up their field trials (forage yield and seed yield trails) and acquire experience with UAV data collection and processing. ILVO will guide partners where necessary. In 2025, breeders will start the actual UAV data collection, and ILVO will develop the models for flower head numbers and biomass. A validation of the model is foreseen in 2026 and 2027.

The BELIS project has received funding from the Horizon Europe research and innovation programme under the Grant Agreement N°101081878.

A diverse collection of pan-European pea field trials established to boost genomic prediction models

A total of 170 pea genotypes from LAMMC, KWS Group, Agritec, CPSBB, CREA, and IAS-CSIC were sown in the field trials at the Lithuanian Research Centre for Agriculture and Forestry (LAMMC). The collection of pea genotypes has been sown by partners in five locations across Germany (KWS), the Czech Republic (Agritec), Bulgaria (CPSBB), and Lithuania (LAMMC). This experiment is part of WP5, which focuses on legume phenotypic and molecular breeding as proof of concept.

Specifically, it addresses Task 5.2 – Breeding of Grain Legumes. This multi-site approach will allow for a comprehensive evaluation and comparison of the genotypes under diverse environmental conditions and provide valuable data for genomic prediction models.

Field trials of BELIS Pan-European Pea at LAMMC.
The BELIS project has received funding from the Horizon Europe research and innovation programme under the Grant Agreement N°101081878.

Faba bean: Use of NIRs for quality traits and validation of markers linked to drought, yield and broomrape resistance

The aims of the Spanish partner IFAPA in BELIS are mainly focused on two actions:

Optimize calibration models using NIRs (Near infrared spectroscopy) for quality traits such as protein, tannins and vicine-convicine content
(Task 3.4).

IFAPA has performed the tannin evaluation in a set of 400 faba bean accessions grown in a new environment (Córdoba, Spain) using the vanillin colorimetric method (Hagerman and Butler, 1978). Measurements of the remaining seed quality parameters (vicine, convicine, and crude proteins) in whole and ground faba beans using HPLC, Spectrophotometry and NIRS is scheduled to start in September 2024.

Figure 1. Measurement of seed quality parameters (tannin content) in faba bean by spectrophotometry analysis.

Validate previous GWAS analyses and develop informative KASPTM markers linked to major traits (drought, yield and broomrape resistance)
(Task 4.3).

In order to validate the significant SNPs for drought detected by GWAS analyses (Gutiérrez et al. 2023), 30 new accessions (4 repetitions each) are being evaluated for morphological, phenological and physiological drought related traits in collaboration with the Julius Kuhn Institute (Germany) in two stationary shelters, one for control and the other for the stress treatment. In the case of the yield a new RIL population (29H x Vf136) has been evaluated this season for seven yield related traits. Data will we used for QTL analysis to reveal possible colocalization/correlation of QTLs with the significant GWAS markers previously identified (Gutiérrez et al. 2024). The validation of markers for broomrape resistance will start October 2024 in collaboration with Agrovegetal.

Figure 2. Left: field trial for drought resistance (June 10, 2024) performed under movable rain out-shelters at JKI (Germany); Right: faba bean field trials (March 19, 2024) for the evaluation of yield related traits in Cordoba (Spain).

The BELIS project has received funding from the Horizon Europe research and innovation programme under the Grant Agreement N°101081878.

UNL-ITQB and IAS-CSIC collaborative effort on phenotyping of Vicia spp.

UNL-ITQB & IAS-CSIC joint activity!

In a collaborative effort, members from the ITQB-UNL team (Portugal) have visit the IAS-CSIC Vicia spp. field trials at Córdoba, Spain, in the past month of April, in a phenotyping marathon of 300 different accessions with their Portable Spectroradiometer Fieldspec4 (Malvern Panalytical).

This equipment will allow to compare the accessions’ spectral signatures in the VIS–NIR–SWIR (350 nm – 2500 nm), which will later be used for the calculation of dozens of vegetation indices associated with the content of chlorophylls, anthocyanins, carotenoids, flavonoids, nitrogen, water, cellulose, lignin, as well as other indices for stress responses and biomass/greenness. These have been the first steps of the establishment of a large scale, high-throughput analysis of Vicia plant´s stress status, achievable through a great international collaboration.

The BELIS project has received funding from the Horizon Europe research and innovation programme under the Grant Agreement N°101081878.

Interview with BELIS Project Coordinator, Bernadette Julier – INRAE

«BELIS is dedicated to enhance genetic progress on legumes through technical novelties as well as improve the conditions to deliver the genetic progress to end-users.« – Bernadette Julier

Introduce yourself, your professional background and your role within the BELIS Project.

I am a geneticist, working at INRAE since 1994. My carrier is dedicated to legume crops, a group of species that is of more and more interest for agriculture sustainability and protein autonomy in Europe. It was already true in 1994 and even earlier but few people put this objective as a top priority at that time. My main interest is on perennial forage legumes, among which lucerne or alfalfa that is adapted to a large number of pedoclimatic conditions. I consider breeding as a lever that is able to fix a number of issues: productivity, quality, resistance to pests and diseases, resistance to cold or drought, adaptation to new growing conditions (in mixtures with other species for example). In combination to agronomic practices, genetics and plant breeding, aiming at producing new improved varieties, contribute to bring progress to the farms and the farmers.

I am the Project Coordinator of BELIS, meaning that I have gathered partners to define the most relevant aspects to be studied, in order to reach the ambition given by Horizon Europe on legume breeding. I am also involved in BELIS as a scientist, engaged in producing more results on the genetic control of key breeding traits in lucerne. In addition, I am working with breeders to test new breeding schemes that make use of molecular markers to speed up genetic progress.

Describe BELIS in 1 sentence.

BELIS is dedicated to enhance genetic progress on legumes through technical novelties as well as improve the conditions to deliver the genetic progress to end-users.

BELIS is made up of 34 partners from 18 countries. Describe this consortium and the role played by the different actors for the development of the project.

The consortium is composed of research institutes, some of them being also breeders, private breeders, and also three technical institutes, one registration office and one international organism in charge of education.

To reach BELIS ambition, we needed skills on different legume species, on different traits, on different technics, on different disciplines.

  • We have chosen to work on many legume species, 7 used at grain legumes for feed and food, 7 used at forage legumes for feed or ecosystem services.
  • The traits under study are related to yield, quality, biotic and abiotic resistances, depending on the species. We are studying new phenotyping methodologies such as remote-sensing technics based on image capture.
  • Technics cover phenotyping, genotyping, detection of markers associated to traits, analysis of genetic diversity, breeding methodologies.
  • Disciplines gather genetics and plant breeding, but also phytopathology, entomology, biochemistry. We have also on board economists and experts of regulations on variety rights.

Why this project is relevant: what challenges is BELIS aiming to respond to?

The European agriculture has a very poor protein autonomy, which means that Europe is importing protein from abroad (soybean meal from South America mainly) and consuming nitrogen fertilizers produced with a huge expense of fossil energy. This situation is a threat for Europe sovereignty. In Europe, the legume crops are under-utilized, mostly because the return for farmers is considered as insufficient. As a consequence, the seed industry is not very active on these species and the efforts dedicated to legume breeding are weak. The genetic gap between legume varieties and the main cash crops is thus continuously broadening.

The challenge of BELIS is to give the conditions to produce a strong genetic progress for legume crops, so that the offer in improved legume varieties is no more a limit to legume cropping.

«We expect that the legume seed sector is able to produce improved varieties for many legume crops, for food, feed and ecosystem services, to be delivered to end-users. This will enhance European autonomy for protein, contribute to crop diversification and reduce energy consumption in agriculture.« – Bernadette Julier

Describe the project’s approach, the type of solutions the project is going to work on, the disciplines involved…

A first approach is to facilitate the phenotypic and molecular breeding, by developing new protocols and testing them in “proof-of-concept” breeding programmes.

A second approach is to improve the variety registration process. We will develop and test cost-effective molecular markers to assist the Distinction-Uniformity-Stability tests. Value for Cultivation and Use tests will be conducted in a few agrizones, at a transnational level, with the objective to enhance the relevance of the evaluations of candidate varieties.

A third approach is to inform about the legume varieties available in Europe and their adaptation to different regions, including by making use to crop modelling.

Another approach is to analyze the governance and business models of the public and private breeders to identify the most promising ones. BELIS will also analyze the financing mechanisms and look for possibilities to include down-stream actors of the value chains in this support.

Finally, BELIS has also the ambition to build a network of all actors involved and interested in legume genetic progress: breeders, scientists, extension services, registration offices, seed industry, and all end-users (feed and food industry, farmer associations, consumer associations). This network, during and beyond BELIS, will be the place for dissemination, trainings, collaborations and all types of exchanges that may boost legume breeding.

BELIS was kicked-off in October 2023. Could you describe what the first steps of the project are?

It was important to all meet in persons and to build a common view of the whole project, in which each one has an important role. Partners discussed about the experiments and studies, focussing on the ones to be installed during the first year.

What may be the project’s benefits for the sector and for society in general?

We expect that the legume seed sector is able to produce improved varieties for many legume crops, for food, feed and ecosystem services, to be delivered to end-users. This will enhance European autonomy for protein, contribute to crop diversification and reduce energy consumption in agriculture.

The BELIS project has received funding from the Horizon Europe research and innovation programme under the Grant Agreement N°101081878.

1st BELIS project Kick-Off Meeting!

The first BELIS project Kick-Off Meeting was held on 17th and 18th October 2023 in Ancenis, France.

This meeting was a great opportunity to present the BELIS objectives and concept: enable a shared understanding of the project and its aims, as well as an effective collaboration between project partners.

The BELIS team also was able to get to know the project partners and foster interaction, establish the basis for working together, as well as present the project organisation, management and decision-making and detail the work packages, timetable and roadmap for the first year of the project.

We also had external presentations from the European Commission by Susana Gaona Saez, Policy Officer from DG AGRI, from the coordinator of the Legume Generation project (our sister Horizon Europe project on legume breeding), Donal Murphy-Bokern, and from Sandra Goritschnig (Alliance Bioversity-CIAT), the coordinator of the Eva-Network project for legume genetic resources characterisation run by CGIAR.

The KoM was the occasion for project partners to visit the facilities of the seed company Cérience, a BELIS partner that runs breeding programmes on forage legumes together with INRAE, and uses the last technologies in seed treatment.

To conclude the meeting, during the working dinner partners presented posters describing their planned activities for breeding each legume species of BELIS, and a vote for the BELIS logo was organised to select collectively the favourite option and share ideas for its improvement.

The BELIS project has received funding from the Horizon Europe research and innovation programme under the Grant Agreement N°101081878.

Launch of Horizon Europe funded project ‘BELIS’: Breeding European Legumes for Increased Sustainability

The 5-year innovation project aims at increasing the competitiveness of the legume breeding industry in the EU and Associated Countries.

BELIS seeks to design conditions that allow an achievement of genetic progress by breeders and its efficient delivery to the seed industry, as well as to other stakeholders.

The BELIS project started on 1st October 2023, with a consortium of 34 partners, including research and technical institutes, plant breeders and seed companies as well as other relevant actors in legume breeding from 18 countries (15 EU countries, Switzerland, United Kingdom and Lebanon).

On 17th and 18th October 2023, the consortium partners will gather at the project’s kick-off meeting that will mark the official launch of this multi-actor project. The event, organised by INRAE as project coordinator, will be held in person and will take place in Ancenis, France, at Cérience/Terrena headquarters. Over these two days, partners will engage in exchanges about BELIS’ objectives and methodologies with the aim of establishing the first steps of the project. The meeting includes a visit to the Terrena seed factory in Beaufort-en-Vallée.

Sustainability and competitiveness challenges in legume breeding

Legumes are widely acknowledged to provide multiple benefits in farming and food systems: They are a source of proteins and other nutrients for human and animal consumption, and when included in cropping systems they can fix atmospheric nitrogen, besides other advantages linked to crop rotations. However, their presence in European cropping systems has not reached its potential, and Europe is still highly dependent on plant protein imports. One of the reasons is the lack of sufficient legume varieties that can provide stable and abundant yields adapted to different agro climatic situations and stresses, with quality traits suitable for food and feed.

Less-than-optimal breeding efforts have so far been invested by public and private breeders. But with adequate breeding inputs, the genetic gains in legume crops could reach those obtained in major crops. While legume breeding is mostly based on traditional phenotypic recurrent selection that requires updating, the use of molecular information and exploitation of genetic resources could also increase selection intensity and speed.

In addition, the genetic progress achieved in new varieties could be delivered much more efficiently to the legume value chain actors (farmers, food and feed industry) and the specific requirements of down-stream actors could be better considered in breeding. The first step of genetic progress delivery relies on the variety registration process, ruled at an international level but conducted by national registration offices. The current registration process needs to be improved by putting a stronger focus on the stakeholders’ needs. The second step is to provide adequate information on the available varieties. Extension services could efficiently deliver information to all actors of the legume value chains, and the seed industry is pivotal in the delivery of genetic progress to farmers by granting access to improved seeds. Finally, improved governance and business models should be developed to allow more efficient, profitable and diverse legume varieties to reach farms.

BELIS objectives

As a whole, the BELIS project will address these challenges and deliver tools and solutions that will enhance the economic sustainability of legume-based farming systems. This ultimate goal will be attained through three specific objectives:

  1. To develop tools and methodologies for cost-effective breeding programmes and deliver proofs of concept, with and for breeders.
  2. To facilitate the economic and regulatory environment: variety registration, variety recommendation and business models.
  3. To implement an efficient, ambitious and durable transfer of innovation through the BELIS platform that includes a network of breeders and actors from scientific research, extension services and seed, food and feed industries, as well as a training portfolio.

The project will focus on seven forage crops (red, white and annual clovers, lucerne, sainfoin, birdsfoot trefoil and vetches) and seven grain crops (pea, faba bean, soybean, white lupin, lentil, chickpea and common bean) representing the diversity of legume species cultivated in Europe.

The project will focus on seven forage crops (red, white and annual clovers, lucerne, sainfoin, birdsfoot trefoil and vetches) and seven grain crops (pea, faba bean, soybean, white lupin, lentil, chickpea and common bean) representing the diversity of legume species cultivated in Europe.

BELIS will provide scientific advances and technical innovations on key aspects of the breeding process: genotyping and phenotyping methods and tools, proposals for agronomic and technological characterisation of varieties, cross testing in different conditions, and pathways for improving and homogenising registration processes. All these results will be streamed to design and validate new governance and business models for the legume breeding industry. An ambitious networking, dissemination, capacity building and exploitation plan will help BELIS results and recommendations reach the breeding industry, the farmers and the regulatory bodies.

BELIS will contribute to a more competitive legume breeding industry in Europe and to delivering profitable and adapted genetic varieties for European farmers seeking better legume seeds.

For more information, please contact:

BELIS Project coordinator: Bernadette Julier bernadette.julier@inrae.fr

Visit BELIS project website: http://www.belisproject.eu/

The BELIS project has received funding from the Horizon Europe research and innovation programme under the Grant Agreement N°101081878.