Syllabus: Plant Cell, Tissue, and Organ Culture –Practical Course

   School Year 2006-2007, Spring Term 2007

   15.3.07       Introduction: Getting Started with Tissue Culture: Conditions of Axenic Culture,
   Laboratory Equipment, Sterile Techniques, Culture Vessels and Different Types of Culture,
   Organization, Protocols, Literature.

   22.3.07       Training of the Work in Sterile Conditions: Sterilization of Tools, Water,
   Vessels, Nutrient Media, Working Area, Methods of the Surface Disinfections of Seeds, Sterile
   Sowing of Seeds, Fern Spores, etc. (Nicotiana, Daucus, Linum).

   22.3.07       Nutrient Medium Preparation: Different Variants of Nutrient Media Solidified
                with Agar (MURASHIGE – SKOOG 1962, GAMBORG et al. 1968, etc.),
                          Preparation and Pouring into Culture Vessels.

   29.3.07       Training of the Work in Sterile Conditions: Subculture of Sterile Culture
                        (Carnivorous Plants: Drosera, Dionaea; Ornamental Plants: Streptocarpus,
   Saintpaulia, Nephrolepis) - Segmentation and Explant Transfer onto Fresh Medium).
   Micropropagation/ De Novo Regeneration: Establishment of Primary Culture, Different Sources of
   Explants - Leaf, Stem and Rhizome (Saintpaulia, Streptocarpus, Begonia, Nephrolepis).

   5.4.07         Micropropagation/ Reproduction: Multiplication by Nodal Segments (Rosa).
   Meristem Culture: Isolation of the Apical and Axillary Meristems of Herbs and Woody Plants
   (Dianthus, Clematis, Rubus),

   12.4.07              Callus Culture: Callogenesis Initiation, Primary Culture of Cambial
   Segments of Carrot Root (Daucus). The Effect of Different Auxin and Cytokinine Ratio on
   Organogenesis of Tobacco Hypocotyl Segments (Nicotiana tabacum).

   19.4.07       A. Induction of Microtuberization on the Potato Nodal Cuttings (Solanum
   tuberosum). B. Culture of Wood Destroyed Fungi: As an Example of the Suspension Culture –
   Maintenance of the Culture and the Inoculum Preparation, (Pleurotus, Lentinus)

   26.4.07       A. Zygotic Embryo Culture: Culture Initiation – Seed Disinfections, Embryo
   Isolation and Inoculation (Picea, Zea, Capsella). B. Somatic Embryogenesis: Direct Somatic
   Embryogenesis Induction – Pea Apical Meristems (Pisum), Flower Receptacles of Cineraria
   (Senecio cruentus – hybrid), Nondirect Somatic Embryogenesis – Carrot Callus (Daucus).

   3.5.07         Cryopreservation: Potato Axillary Meristem Isolation, Encapsulation of Isolated
   Meristems into Alginate Beads, transfer of beads to M-S medium, (Dehydration and Freezing of
   Beads in Liquid N2).

   10.5.07       A. Phytopatology in vitro, Indexing for Microbial Cultivable Contamination of
   Primary and Long-term Culture, Test of Solanum tuberosum a Nicotiana tabacum Culture Health.
   B. Acclimation of Micropropagated Plants.Evaluation of Experimental Results and Control of
   Protocols, Test.

   17.5.06       Excursion to the Plant Tissue Culture Lab of Mendel Agriculture and Forestry
                           University and credit.