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Improving the methods for cryopreservation of endangered conifers
Gaiduschová, Daniela
Preservation of plant genetic material is essential for any biological field. Compared to serial in vitro sub-culturing, cryopreservation often represents optimal long-term conservation technique. The crucial factors influencing successful cryopreservation include 1) proper pre-treatment, 2) cryoprotectants, 3) adequate freezing pace and 4) cold-hardening. Finding the appropriate method for cryopreservation of each species is the ultimate goal of research in this field. This work is focused on finding the optimal protocol for cryopreservation of embryogenic cell masses of coniferous species bog pine (Pinus uncinata subsp. uliginosa) and optimizing the existing one for European silver fir (Abies alba), both endangered in the Czech countryside. Cryopreservation design compares effectivity of different pre-treatments and cryoprotectants, determines the role of cold-acclimation step and evaluates resistance of selected conifer cell lines to freezing and desiccation. Post-thaw recovery of Bog pine cell line BE4 was better than cell line BR1, where no living cells were visible using FDA/PI staining. Proliferation rate was higher for European silver fir than for bog pine, especially for cell line II-2-10. Cryoprotective mixture with DMSO (PGD I) proved to be more effective than the mixture with glycerol (PGG) in cryopreservation of European silver fir. PGD I also proved to be optimal cryoprotective mixture for bog pine. In both cases, 0.4M sucrose was used as a pre-treatment and cold-hardening was included.
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Improving the methods for cryopreservation of endangered conifers
Gaiduschová, Daniela
Preservation of plant genetic material is essential for any biological field. Compared to serial in vitro sub-culturing, cryopreservation often represents optimal long-term conservation technique. The crucial factors influencing successful cryopreservation include 1) proper pre-treatment, 2) cryoprotectants, 3) adequate freezing pace and 4) cold-hardening. Finding the appropriate method for cryopreservation of each species is the ultimate goal of research in this field. This work is focused on finding the optimal protocol for cryopreservation of embryogenic cell masses of coniferous species bog pine (Pinus uncinata subsp. uliginosa) and optimizing the existing one for European silver fir (Abies alba), both endangered in the Czech countryside. Cryopreservation design compares effectivity of different pre-treatments and cryoprotectants, determines the role of cold-acclimation step and evaluates resistance of selected conifer cell lines to freezing and desiccation. Post-thaw recovery of Bog pine cell line BE4 was better than cell line BR1, where no living cells were visible using FDA/PI staining. Proliferation rate was higher for European silver fir than for bog pine, especially for cell line II-2-10. Cryoprotective mixture with DMSO (PGD I) proved to be more effective than the mixture with glycerol (PGG) in cryopreservation of European silver fir. PGD I also proved to be optimal cryoprotective mixture for bog pine. In both cases, 0.4M sucrose was used as a pre-treatment and cold-hardening was included.
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Evaluation of taste repellents for protection of tree planting stock from deer browsing
Triner, Libor ; Kuneš, Ivan (advisor) ; Kupka, Ivo (referee)
The main goal of this thesis is to assess the efficacy of two repellent products with active ingredients denatonium benzoate and capsaicin to protect the newly established forest tree planting prior to browsing damage by wild game. Repellents were tested on two different experimental plots planted with silver fir (Abies alba). Application of repellents held in three terms and total three times were the damages, which were included in the framework of the six-rating scale, has been detected. The effectiveness of these repellents has been demonstrated already during the second term damage assessments caused by game. During the more than 14-month experiment was damaged almost 2.5 times fewer seedlings treated with repellents based on denatonium benzoate and capsaicin than untreated seedlings, serving as a control. The most common form of damage to the tree was bitten off terminal bud with little scope of additional damage. Damage to seedlings by game differed significantly even within the investigated areas. Use of repellents did not affect the physiological death of seedlings. It was justify the conclusion that most of the damages were caused by roe deer (Capreolus capreolus). Based on several scientific papers on the efficacy of repellent products based on capsaicin and denatonium benzoate was concluded that the main active ingredient in tested repellents is capsaicin.
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Studium vlivu fytohormonů a ektomykorhizních hub při maturaci a konverzi somatických embryí jehličnanů
Filippova, Mariia
This thesis deals with the exploration of the influence of various phytohormones, primarily the influence of abscisic acid on the process of maturation and the influence of auxin and gibberellin on the process of germination of two types of conifers, Abies alba and Picea abies, often used as a material in the discovery of somatic embryogenesis. Each genotype prefers specific concentration ABA. 15 uM and 45 uM is the most effective for the cell lines Abies alba; 15 uM and 30 uM ABA for Picea abies. Auxin (IBA) and gibberellin (GA3) did not support the growth of roots of Abies alba. The interaction between fungal cultures Paxillus involutus and Amanita muscaria and between the embryos during the germination was studied as well. Fungal culture Paxillus involutus had very fast speed of growth on the germination medium. Fungal culture Amanita muscaria was growing very slowly. The effect of fungi on the germination has not been observed.
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