| Abstract [eng] |
The purpose of this work was to evaluate the effect of Stevia rebaudiana Bertoni natural stevioside sweetener on in vitro hybrid aspen cultivation. Four main objectives have been formulated: to evaluate stevia plant growth rate in different in vitro media and lighting; to form the most effective nutrient media for hybrid aspen microshoots using a stevioside sweetener; to select the most informative oligonucleotide primers for stevia DNA analysis; to evaluate DNA identity of the Stevia rebaudiana Bertoni clones using selected primers. Methods used for testing: producing nutrients by substituting a stevioside sweetener as a carbon source; isolation of DNA using CTAB method; DNA multiplication (PCR) using random primers; electrophoresis separation of PCR products, gel analysis and documentation of results. After evaluating the growth rate of stevia plants, we found that planted 20 mm cuttings after 2 weeks of growth in hormonal and non-hormonal media grow twice or even tripled at 2570 luxlight. The most effective nutrient media for hybrid aspen production were obtained using a mixture of 0.06 g/l sucrose and 0.06 g/l white stevioside in the medium. After three weeks of growing in such a medium, the hybrid aspen microshoots reach a height of 29 7.53 mm compared to the control microshoots that grow to 21.38 5.34 mm height. The most informative oligonucleotide primers for selected stevia DNA analysis were: Roth 170 - 01, Roth 170 - 02, Roth 170 - 03, Roth 170 - 04, Roth 170 - 05, Roth 170 - 10, Roth A - 14, Roth A - 19, Roth B - 01, Roth B -08, Roth B - 15, Roth B - 17. 100% polymorphism was obtained by determining the DNA identity of stevia plants and their clones. |
