The adaptation of regenerant plants to environmental conditions is the final stage of micropropagation. According to a number of authors, when in vitro plants are transferred to in vivo non-sterile conditions, a significant percentage of mortality is recorded. In a previous publication, the regenerative capacity of strawberry (Fragaria vesca L.) in vitro tissues on MS culture medium (Murashige & Skoog, 1962) and a regenerants was obtained (Chornobrov O. Yu., 2019). The objective of the study is to develop an optimal protocol of acclimation of in vitro F. vesca plants to in vivo conditions. Methods. Biotechnological and statistical methods of research were applied. For the research 'Ruiana' and 'Zhovte Dyvo' cultivars were used with in vitro cultivation cycle of 30-35 days. Prepared plants were planted in 0.33 L plastic contai ners, one piece in a mixture of coconut substrate and perlite (3:1). Plants were kept under high relative humidity (85-90%) conditions for 3-5 days, 6-8 days and 10-14 days. The studies were carried out in the Plant Biotechnology Laboratory of SS of NULES of Ukraine "BFRS" during 2019-2020. Results. The duration of Fragaria vesca regenerant plants exposure in conditions of high relative humidity significantly affected adaptation efficiency. The proportion of 'Ruiana' and 'Zhovte Dyvo' plants adapted to the greenhouse conditions were 47.6 ± 2.5% and 60.0 ± 1.7%, respectively, when the plants were kept for 10-14 days. A significant efficiency of plant adaptation (more than 70%) was obtained under condition of preliminarily exposure the roots of the plants in an auxin solution for 25-30 minutes with daily application of 30% solution of glycerine as foliar spray. The plants adapted to the greenhouse conditions had pigmentation characteristic of the variety, without signs of chlorosis and vitrification. Conclusions. An optimal protocol for in vitro adaptation of F. vesca cultivars to in vivo conditions was developed and viable plants were obtained. Further research will be aimed at studying the growth and development of F. vesca regenerant plants in open ground.
One of the methods of obtaining planting material of deciduous plants, in particular common ash (Fraxinus excelsior L.), broad-leaved linden (Tilia platyphyllos Scop.) and silver birch (Betula pendula Roth) is microclonal propagation. Asepticity of explants is a prerequisite for microclonal plant propagation. Chemical sterilization with liquid substances is mostly used for this purpose. The mode of decontamination is influenced by a number of factors, in particular the genotype of the plant. The purpose of the study was to optimize the sterilization protocol of F. excelsior, T. platyphyllos and B. pendula explants for microclonal propagation. For research, 20–30 cm of shoots isolated from 12-year-old T. platyphyllos, 10-year-old B. pendula, and 15-year-old F. excelsior in February-March 2021 were used. Plant material was cultured according to conventional methods on a nutrient medium MS (Murashige & Skoog, 1962). Biotechnological methods were used (plant tissue culture in vitro, microclonal propagation). MS Excel software package was used to process the experimental data, the mean and its standard error were calculated. One-way analysis of variance (ANOVA) was performed to analyze the effect of explant sterilization on asepsis. The expediency of keeping plant material during the day in 0.1 % solution of «Samshit» F. excelsior and 0.3 % solution «Fundazole» T. platyphyllos and B. pendula is shown. The sterilization protocol of experimental plants (efficiency over 50 %) was optimized by using a stepwise method using 70 % ethyl alcohol, 1.0 % and 2.0 % AgNO3 and 2.5 % and 5.0 % NaClO. The effect of the sterilization regime of experimental plants on asepsis is statistically significant at the level of 5%. To initiate the explants, a culture medium according to the MS prescription was used with the addition of 0.25 mg/L kinetin and 2.0 g/L activated carbon. Further studies are aimed at developing a protocol for direct regeneration of microshoots of F. excelsior, T. platyphyllos and B. pendula under the action of components of the culture medium in vitro.
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