TECHNOLOGIES
Pathogens and Pest BIOASSAYS TO MINE GENETIC DIVERSITY
To untap the potential of genetic diversity, we are developing high-throughput bioassays. The TCI Laboratory has state-of-the-art facililities to conduct high-throughput screening for virus resistance. It includes the development of infectious clones and in vitro bioassays to assess resistance against pathogens and their insect vectors.
Growth room with cages to conduct bioassays with insect vectors on crop species.
Development of infectious clones combining assemblies in bacteria and yeast. (Shakir et al., 2023)
SEQUENCING TECHNOLOGIES TO CHARACTERIZE DIVERSITY AND EVOLUTION
Long Read sequencing technologies have revolutionized the field of plant and pathogen biology by increasing the quality of large crop genomes. They can also be used to generate single read full-length genomes of pathogens with small genomes such as viruses. The TCI Laboratory has established methods to monitor the evolution of DNA virus populations in disease resistant plants. This tool is used to assess the durability of disease resistance. The Cider-Seq method is also suitable to profile transposable elements in plants.
General pipeline of the Cider-Seq method to profile ssDNA viruses and extrachromosomal circular DNAs (eccDNAs) (Adapted from Metha et al., 2020)
GENOME EDITING
Genetic transformation and genome editing have emerged as important techniques for both fundamental and applied plant biology. The TCI Laboratory has established high-throughput methods to genetically transformed crop plants. Lately, those methods have been completed with approaches to generate T-DNA-free genome editing events. Those approaches are particularly suitable to develop varieties for farmers and industries.
The genetic transformation and editing platforms available at TCI Laboratory are used to valide candidate genes (genes associated with biotic or abiotic stress resistance) in both model and crop plant species.
Overall scheme for the generation of transgenic and genome edited banana and cassava with the TCI platform (Zorrilla et al., 2020)
CRYOPRESERVATION OF TROPical plants
The TCI Laboratory has a long and proven expertise in the development of protocols to saveguard vegetatively propagated plant species. State-of-the-art cryotanks allow theTCI Laboratory to have a storing capacity of several thousands genotypes.
SCreening for Drought Stress tolerance
The TCI laboratory uses a so-called Bananatainer and state-of-the-art greenhouse facilities to screen the banana collection for capacity to cope with water deprivation.
Biological nitrification inhibition (BNI)
Some plant species, including several tropical plant species, have the capacity to inhibit key steps in the nitrogen cycling occuring in the soil. Root exudates from certain genotypes of sorghum, brachiaria, wheat and other species have been demonstrated to contain molecules inhibiting the nitrification processes converting ammonium into nitrates. Because nitrogen losses represent a major pollution to the environment, there is a growing interest to identify plant species which can mitigate or slow down the nitrification and denitrification in the agrosystems.
A platform to screen BNI activities in plant tissues and exudates has been developed in order to identify species and genotypes of interest as well as to study the molecular mechanisms of nitrification inhibition. This work is part of the expertise available in the Plant Genetics and Rhizospheric Processes at University of Liège.
Overview of the pipeline for identification of biological nitrification inhibition (BNI) capacity in wheat roots. (a, b) Wheat plants growing in the large hydroponic tanks under glasshouse conditions; (c, d) high-throughput root exudate collection system; (e) pure anoxic culture of the wild-type Nitrosomonas europaea; (f) concentration process of root exudates with the rotary evaporator; (g) microplate bioassay after Griess staining; (h) timeline for sample processing and analysis.(Jauregui et al., 2023)
BIOPESTICIDE SCREENING
The TCI Laboratory uses in vitro bioassays to screen and identify natural molecules with pesticide activities with a focus on virus vectors (i.e. whitefly, aphid). A platform using artificial feeding assays and in planta bioassay enables testing plant extracts, proteins and other selected molecules for activities against different developmental stages of the insect vectors. The platform can also be used to identify insect proteins interacting with the viruses they vector.
Articial feeding bioassay at TCI Lab