In Vitro Lab
by Prof. Stefaan Werbrouck
Our
Research
Plant tissue culture offers a controlled environment for plant propagation, genetic modification, and the production of valuable secondary metabolites. However, several challenges limit its efficiency and success. These challenges include slow regeneration rates, susceptibility to viral and bacterial contamination, and limited production of desired compounds. Our research addresses these challenges through the 5 following research domains:
5 Research Domains
1. Plant Growth Regulators
Meta-topolin derivatives for good quality micropropagation
Cytokinin selection is crucial for successful plant micropropagation. These hormones exhibit a complex range of stimulatory and inhibitory effects on plant development. Naturally occurring cytokinins include both isoprenoid compounds and diverse aromatic derivatives. Our research explores the potential of novel topolin-type cytokinins to enhance tissue culture techniques in various plant species. This work aims to overcome specific challenges encountered during the micropropagation process.
Fluorated Meta-topolins
Fluorine modification can enhance molecular stability and potency. We investigate fluorine-modified topolin derivatives as powerful cytokinins, demonstrating exceptional results in Phalaenopsis micropropagation. These halogenated compounds significantly increase shoot formation in Phalaenopsis hybrids, which typically exhibit slow in vitro growth. Interestingly, container aeration influences the cytokinin's effects on shoot growth.of Phalaenopsis.
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Cytokinin Oxidation Inhibitors
Low plant regeneration after transformation hinders progress in genetic engineering. Our research tackles this by developing efficient micropropagation systems. We utilize novel, designed cytokinin oxidase inhibitors for various economically important crops. This collaborative effort involves the Institute of Experimental Botany in Czech Republic.
2. In Vitro Technology
LEDs as unconventional light sources
LEDs are being explored as an alternative light source for plant micropropagation due to their efficient energy use and ability to control plant development through specific wavelengths. Our research utilizes growth chambers with red, blue, and far-red LEDs alongside production LEDs to study the impact of these monochromatic light spectra on plant morphology and development in vitro.
Slow release nanoparticles of plant growth regulators
Gradients of growth regulators are crucial for in vitro plant regeneration. We address this by developing a system using calcium carbonate particles embedded with regulators, further aggregated with calcium alginate. These milliparticles, containing millions of microscopic carriers, are used with explants of difficult-to- propagate plants. This approach facilitates controlled release and allows observation of cell dedifferentiation into complete plantlets.
Temporary Immersion Bioreactors
The SETIS™ temporary immersion system combines a plant growth chamber with a connected nutrient reservoir. Periodic, short immersions in the nutrient medium and regular headspace aeration promote rapid in vitro growth for various plant species. Our research evaluates the advantages and limitations of the SETIS™ system, applying it to large-scale plant propagation experiments.
3. Plant Health
Fungal volatiles as biostimulants
Serendipita indica is a endomycorrhiza fungus. We study the impact of its volatile organic compounds (VOCs) on salt-stressed plants in vitro, using both a semi-solid medium and a temporary immersion bioreactor system (SETIS). VOCs improved plant growth under all salt levels and even without added stress. Increased antioxidant activity suggests VOCs help plants cope with stress. The adapted SETIS system will be valuable for further research on volatile-compound mechanisms.
Virus free mother plant stocks
Viral diseases threaten yields in vegetatively propagated crops, creating challenges for propagation companies that supply the agricultural sector. Traditional virus detection methods are limited, but high-throughput sequencing (HTS) offers a rapid and comprehensive way to identify the full "virome" of a plant. We develop and optimize HTS-based virus detection for certification and sanitation therapy, allowing companies to produce virus-free plants. Additionally, we explore HTS for detecting bacteria and other pathogens, ultimately enhancing crop health and strengthening the market position of propagation companies.
Identifying and beating endogenous bacteria
Internal bacteria contaminate in vitro cultures, hindering plant growth and valuable compound production. Disrupting hormone balance, these bacteria pose a challenge even when dormant. Our research explores how bacterial communities affect in vitro plant health. We use isolation and DNA techniques to identify contaminants, aiming to develop strategies for both eradication and potential exploitation.
4. Secondary Metabolites
Callus and cell cultures
While stilbenes like resveratrol offer health benefits, their production is limited and cannot meet the market demand. This study investigates boosting stilbene production in peanuts using in callus and cell suspension cultures. Since callus cultures yield low amounts, we are testing fungal-derived biotic elicitors to increase the synthesis of resveratrol, piceatannol, and piceid.
5. Ploidy Breeding
Polyploidy, the doubling of a plant's genome, is a common occurrence in plant evolution and has been widely utilized in agriculture and horticulture to enhance crop traits. Despite the potential benefits of increased cell size, biomass, and stress tolerance, the application of polyploidy in forestry has been limited. However, polyploid trees offer promising advantages, such as larger leaf area, improved drought resistance, and potential applications in tree breeding programs. This approach is being explored on a case-by-case basis for various tree species, and our research relies on the experience to regenerate adventitious shoots from tissue treated with mitosis inhibitors like oryzalin.
6. Screening Services
As part of the Cropfit network, we collaborate with colleagues contributing their innovative plant bioassays to evaluate the next generation of sustainable crop protection agents, fertilizers and biostimulants from both academic and industrial sources. Our role within this collaboration involves testing new compounds for their hormone-like ativity in a series of bioassays.
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