Research Projects

Research Organisation Partners

• Kemijski inštitut, Hajdrihova ulica 19, 1000 Ljubljana
• Institut “Jožef Stefan”, Jamova cesta 39, 1000 Ljubljana

Abstract

Cannabis sativa L. is an economically significant crop with applications in pharmaceuticals, cosmetics, food, and sustainable industries. However, its genetic improvement remains slow due to the limitations of traditional breeding methods. F1 hybrid varieties offer advantages such as heterosis, uniformity, and enhanced traits, yet their development is hindered by the lack of completely homozygous parental lines. This project, HaploCAN, aims to revolutionize C. sativa breeding by developing haploid inducer lines (HILs) for the efficient production of haploid and doubled haploid (DH) plants. These lines will accelerate the breeding of both medical cannabis and industrial hemp, facilitating the creation of elite F1 hybrids with desirable traits.

To achieve this, we will employ two complementary approaches: CRISPR/Cas9 genome editing and irradiation-induced mutagenesis. Genome editing will be used to introduce targeted changes in genes critical for haploid induction, leveraging state-of-the-art editing techniques, including CC-Exo and CC-PE systems. Simultaneously, gamma irradiation will induce random mutations to identify alternative genetic pathways for haploid induction, circumventing GMO regulations in regions with restrictive policies.

The project is structured into five work packages: (1) bioinformatics analysis and plasmid development, (2) genome editing of C. sativa, (3) mutagenesis via irradiation, (4) haploid induction through crosses with HIL, and (5) dissemination, risk assessment, and management. The work will be conducted in collaboration with leading Slovenian research institutions: the Biotechnical Faculty of the University of Ljubljana, the National Institute of Chemistry, and the Jožef Stefan Institute. Additionally, we will collaborate with international partners to optimize in vitro regeneration protocols and genetic transformation efficiency.

HaploCAN's expected outcomes include the establishment of the first haploid inducer lines in C. sativa, the first successful production of DH C. sativa plants, and a comparative evaluation of genome editing versus irradiation for mutation induction. These findings will be disseminated through high-impact publications, international conferences, and industry collaborations. The project’s broader impact extends to academia, breeding companies, the medical cannabis and industrial hemp sector, enabling faster and more precise genetic improvements. By bridging the gap between traditional and modern breeding technologies, this research will position C. sativa at the forefront of advanced plant breeding strategies, supporting its expansion in sustainable agriculture and biotechnology.

External link to Researchers Open in new window

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