New cells growing on the former ZE either form calluses of different patterns and colours or differentiate to suspensor and meristem cells of early somatic embryos founding new embryogennic culture.
Our research group is mainly involved in the studies of the metabolism and physiological functions of growth regulators, polyamines and phenolic compounds in plants. We investigate the role of these biologically active compounds in plant development and in the response of plants to abiotic stresses.
In our experiments we use the diverse plant systems from the whole plants to the cell cultures. Our research is primarily focused on somatic embryogenesis of conifers. In the scope of this theme we study the regulation of somatic embryo development, the role of phytohormones in somatic embryogenesis and the effects of abiotic stresses on somatic embryos.
We use a wide array of approaches:
Microscopy – light, confocal and electron microscopy, enhanced by advanced computer image analysis
Biochemical methods – studies of activities of enzymes involved in metabolism of biologically active compounds (e.g. radiometry)
Molecular biology methods – specific gene expressions, and transformation of tissue cultures
Analytical methods – qualitative and quantitative determination of biologically active compounds by gas- and liquid chromatography in tandem with mass spectroscopic detection (cooperation with the IEB Laboratory of mass spectrometry).
New somatic embryogennic cultures can be induced from zygotic embryos cultivated in vitro on the induction medium. Phytohormones auxins and cytokinins play important role in this phase.
Somatic embryogenic culture can be induced from immature zygotic embryos extirped from seeds in the course of summer.
Quantification of histochemical detection of polyphenolic compounds in somatic embryos of Norway spruce
Under standard cultivation conditions, Norway spruce somatic embryos (SEs) accumulate polyphenolic compounds mostly in the root cap. In response to UV-B, the accumulation of polyphenolics increases in protodermal cells and subprotodermal cortical cells and idioblasts in hypocotyl and cotyledons where they can attenuate UV-B stress. To quantify polyphenolic compounds accumulated in somatic embryos on the histological level resin sections were prepared, stained with Toluidine Blue.