Plants of Schlumbergera truncata (Haw.) Moran were obtained by indirect morphogenesis from the segment section of shoots in vitro, they were multiplied and rooted. Also were determined the effect of the lighting regime, the composition of the nutrient medium on the consistency and frequency of callus formation. The studies were conducted during 2016–2018. The mode of effective sterilization (more than 90%) of S. truncata plant explants using 0.1% HgCl2 for 7–8 min was established. Optimal conditions for the induction of callus formation in stem node segments of S. truncata plants (rate more than 90% and significant growth) were created on MS (Murashige and Skoog 1962) nutrient medium supplemented with 1.0 mg/l BAP (6-benzylaminopurine) and 0.3 mg/l NAA (1-naphthylacetic acid) under conditions of placement on the nutrient medium and doing a significant number of cuts on the explants. The light intensity of 2.0–3.0 klx, obtained by a callus of dense consistency of dark green pigmentation, when using the thermostat condition without illumination, the callus had loose consistency, dark yellow pigmentation. It is established that the influence of the lighting regime and the composition of the nutrient medium on the frequency of callus formation is statistically significant. The largest number of shoots was obtained on the MS medium with the addition of 2.0 mg/l of BA. At the same times, shoot proliferation and root induction in such numbers were observed on MS culture medium with the addition of 0.5 mg/l BA and 0.5 mg/l kinetin (multiplication factor – 8.8±0.6 per 60-day cultivation cycle).
Conservation and reproduction of rare genotypes of Salix L. species, in particular the blunt-leaved willow (Salix retusa L.) and Jacquin’s Willow (Salix alpina Scop.) that are listed in the Red Data Book of Ukraine in the status of rare and endangered species, is one of the urgent tasks of the present. The aim of research was to develop methods of introduction of S. retusa and S. alpina into in vitro culture for their mass reproduction and conservation. The plant material was cultivated on a culture medium prescribed by MS, WPM, and DKW with the addition of growth regulators according to the conventional method. Effective sterilization (over 80%) of explants of S. retusa and S. alpina was achieved by applying a stepwise method, which consisted of consistently maintaining them in solutions 0.1% HgCl2 and 1.0% AgNO3 for 5–6 min. Significant results in the regeneration of explants by activating the growth of available meristems in vitro were observed on MS with the addition of 0.25–0.5 mg/l 6-(Furfurylaminopurine, kinetin) and 2 g/l activated carbon. Our further research will serve as a base for developing microclonal propagation of S. retusa and S. alpina for their conservation and reproduction in vitro.
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.
European white elm (Ulmus laevis Pall.) tissue in vitro is a donor material for obtaining cultures with stable resistance to pathologies of infectious origin, namely to Dutch elm disease. To solve this problem, it is necessary to develop an effective protocol for the regeneration of U. laevis in vitro. The purpose of this study is to investigate the effect of chloramine concentrations on the mycobiota of U. laevis plant tissues for propagation in vitro. 10-15 cm parts of shoots from 25-year-old U. laevis were used as plant material. The study was conducted in the autumn of 2021. Microshoots previously sterilized with chloramine (1.0%, 2.5%, 5.0%, 10.0%) for 10 min were cultivated on a solid nutrient medium according to the WPM recipe (McCown & Lloyd, 1981) with the addition of 0.2 mg∙l -1 2 - iP (6-(γ,γ- Dimethylallylamino)purine) and 2.0 g∙l-1 of activated carbon. For microbiological analysis, sterilised plant material was cultured by accumulation in Petri dishes with a nutrient medium (sour potato agar) in a thermostat without lighting at +26 ± 1°C and a relative humidity of 68 ± 2%. Methods of biotechnological, mycological, and statistical research were employed in this study. Over 95% of the samples were found to be infected with microscopic fungi of the genus Mucor Fresen., Penicillium Link, Chaetomium Kunze and Trichoderma Pers. The effect of preparation concentration on the total number of infected explants is statistically insignificant at 5%. It was found that 5.0% preparation is effective for neutralising mycobiota of the genus Chaetomium and Trichoderma; 10.0% – for neutralising Penicillium mycobiota. If the concentration of chloramine increases, the intensity of infection of explants with mycobiota of various genera decreases. As a result of the research, a small amount of aseptic cultures were obtained from the shoots of U. laevis isolated in autumn. This study is relevant for biologists, biotechnologists, microbiologists, and biological scientists
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