Description of the research
The scientific work in the field of plant biotechnology and experimental biology focuses on the following research topics:
- Analysis of the plant genome structure and evolution at the cytomolecular level.
This research includes analyses of the chromatin structure during the different phases of the cell cycle, analyses of the nuclear genome size in different species and determining the physical localisation of both genes and non-coding DNA sequences, including the genes encoding the important traits in crop plants. The research also focuses on the detection and characterisation of the chromosomal changes that occur during both the evolution and ontogenesis, as well as being the result of biotechnological modifications. The stability of a plant genome after treatment with physical and chemical mutagens is also assessed. This research is often conducted on model species, increases its complexity and interdisciplinary character.
- Functional genomics of crop and model plants.
The main field of research includes studies on identification and determining the role of genes related to the processes of plant development in vivo and in vitro, as well as genes controlling plant response to abiotic stresses. The research is carried out on a crop species barley (Hordeum vulgare) and a model species Arabidopsis thaliana. The studies in barley include identification and regulation of genes which are involved in: (1) root system development with the special emphasis on morphogenesis of root hairs, (2) metabolism of brassinosteroids and strigolactanes, the plant growth regulators that affect shoot development, (3) response to abiotic stresses, especially water deficit, (4) DNA repair. The research on deciphering the role of epigenetic modifications, especially DNA methylation, in the adaptation of plants to environmental stress conditions is also conducted. A separate subject of studies includes analysis of genetic and epigenetic mechanisms engaged in plant regeneration in vitro. These mechanisms are studied in A. thaliana in the aspect of identification of major genes controlling developmental plasticity of plant cells and elucidating their regulation at the transcriptional and post-transcriptional level. In barley, the molecular and cytological processes that are involved in the phenomenon of albinism in in vitro microspore culture are being determined.
- Morphogenetic analyses and mathematical modelling of plant organ growth and development.
The research focuses on the regulation of plant organ and tissue morphogenesis with special emphasis on the apical meristems of the shoot and root and the formation of lateral organs. Modelling and quantitative analysis of organ growth and geometry are complemented by biomechanical studies of growth regulation, including the role of the cytoskeleton and the mechanical properties of the cell walls in the regulation of the functions of the apical and lateral meristems. Mathematical growth models that include oriented cell divisions and computer simulations are based on empirical data. Such an approach enables the identification of the biomechanical phenotype traits that are important for breeding crop plants.
- Genesis and the function of the electrical potential in plants
The generation of electrical gradients is a fundamental aspect of signal transduction in plants. The studies concern the changes in the plasma membrane potential and the modulation of the slow vacuolar (SV) channels in response to light, temperature, heavy metals, hormones and other agents. Two basic techniques are used in the experiments – the patch-clamp technique and impaled microelectrodes.
- Analysis of symplasmic communication and physico-chemical changes in the cell wall during cell differentiation.
The mechanisms of plant cell differentiation are studied in terms of symplasmic communication and changes in the physico-chemical properties of the cell wall during plant development. Investigations of symplasmic communication relate to analyses of the symplasmic domains and their role in plant development from embryo to seedling and during the formation of tissues and organs, as well as to the changes that are caused by environmental conditions. Moreover, the studies concern analyses of the changes in the chemical components of the cell wall (cellulose, hemicellulose, pectins and AGPs) during cell differentiation and under the influence of biotic and abiotic stress factors. These types of analyses can contribute to a better understanding of the role of the symplasm and apoplast in plant development and hence the future use of this knowledge to “design” a plant that has the required features.
The studies use the following methods:
- imaging and flow cytometry
- molecular cytogenetics techniques, e.g. fluorescent DNA-DNA hybridisation in situ, immunostaining techniques
- genotoxicity assays (TUNEL, comet method)
- techniques of 3D imaging using confocal microscopy and advanced methods of digital image processing
- genome editing and targeted mutagenesis (CRISPR/Cas9 system)
- transmission and scanning electron microscopy
- molecular markers (AFLP, SSR, SNP)
- DNA cloning and sequencing, including NGS;
- transcriptome analysis using DNA microarray, Real-Time qPCR; RNA-seq
- differential proteome analysis;
- TILLING strategy;
- immunocytochemical methods for detection of cell wall chemical components usingthe light and electron microscopy;
- application of low- and high-molecular weight fluorochromes and dextrans for symplasmic communication analysis
- bioinformatics techniques;
- plant in vitro cultures, including anther, isolated microspore, and protoplasts culture;
- genetic transformation techniques using Agrobacterium;
- classical electrophysiological measurements patch clamp technique cell pressure probe technique
- tensometer investigations and rheological tests
- computer modelling of plant growth and development using the growth tensor method and biomechanical approach