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Gymnadenia conopsea (Fragrant orchid) is a widespread model species for the study of polymorphism in the European part of Russia and abroad. Flow cytometry refers to rapid methods effective for assessing the structure of a population by DNA size estimation. However, this method is only available in habitats with close proximity to the place of the experiment, because this indicator is determined only in fresh leaves. A small population of G. conopsea in the industrial area adjacent to the Novosibirsk urban agglomeration in the Novosibirsk Region (NR) was studied. Only diploids were detected in the G. conopsea cenopopulation occurred in the industrial zone of the NR. In this study we pointed on significance of genome size variation, which correlates with the adaptive evolution of the G. conopsea in West Siberia. The revealed absence of minor cytotypes (individuals with a higher level of ploidy) in G. conopsea population reduces intraspecific and intrapopulation diversity of the species.

In this study the results of agrobacterial transformation of Astragalus membranaceus (Fisch. Ex Link) Bunge, a medicinal plant from Mongolia and China with a wide spectrum of pharmacological action were presented. As explants for transformation, in vitro derived microshoots as well as hypocotyls, cotyledons and primary shoots isolated from seedlings were used. The most effective from two tested Agrobacterium rhizogenes strains (A4-RT, 15834 SWISS) was 15834 SWISS, causing the transformation of various types of explants, including primary shoots as a more responsive type. The optimal co-cultivation time of explants with bacterial suspension (48 hours) and the highest transformation rate of primary shoots (36.4%) were established. PCR analysis confirmed the transgenic nature of the roots transformed with strain 15834 SWISS and the absence of bacterial contamination.

In the West Siberia there is very little data on the biology of cenopopulations of Gymnadenia conopsea. Habitat destruction and environmental changes, which potentially affected species long-term viability, led to G. conopsea cenopopulations fragmentation and isolation. A detailed study of the morphology variability of the Fragrant orchid representatives was carried out in the cenopopulation in Novosibirsk region. Our results indicate that asymbiotic germination of G. conopsea seeds is difficult to achieve and the species has serious conservation issues. It is the first report when the introduction of G. conopsea in tissue culture was undertaken as the initial stage of germplasm conservation of Siberian cenopopulation. Three modified nutrient media with different growth additives were tested to promote protocorm and seedling formation. The advanced G. conopsea seedlings establishment was obtained in 1/3 Murashige and Skoog (MS) medium, supplemented by 1.0 mgl−1 2-isopentenyladenine (2iP), 0.1 mgl−1 1-Naphthaleneacetic acid (NAA) and 10% coconut water. This study allowed establishing a reliable and reproducible system for the G. conopsea maintenance and conservation ex situ.

For the first time light and scanning electron microscopy have been to reveal changes in the anatomical structure of leaf blades of Fragaria × ananassa Duch. regenerants cultivated in nutrient media supplemented with biogenic silica and green-tea-flavonoids-based mechanocomposite. An increase in the density of stomata per unit area of the leaf, size of the stomatal gaps, thickness of the epidermis and mesophyll have been noted. The revealed changes in the leaf blade structure under mechanocomposite treatment contribute to the further successful acclimatization of regenerants to ex vitro conditions.

Rhododendron mucronulatum Turcz., distributed throughout the northern region of East Asia has been considered to be an alternative natural source of taxifolin (dihydroquercetin) and rutin. The present study was conducted based on a biotechnological approach to develop an environment friendly and efficient system to produce taxifolin and rutin in R. mucronulatum microshoots, using different thidiazuron (TDZ) treatments (0.1; 0.5; 2.5 µM) in combination with various types of lighting including fluorescent (FL) and light-emitting diode (LED) (R/B- 80% red + 20% blue; 5LED-20% red + 20% blue + 20% green + 20% yellow + 20% white). The highest number of shoots per explant was obtained under 0.5 µM TDZ combined with 5LED in comparison with FL lighting. Among shoot clusters obtained under different lighting types and TDZ concentrations, a considerable increase in fresh and dry weight was observed in ones cultivated on medium, supplemented with 2.5 µM TDZ under FL and 0.5 µM TDZ at R/B or 5LED. The content of total chlorophylls in R. mucronulatum microshoots increased on TDZ-free medium under FL lighting, whereas, the TDZ treatment decreased chlorophylls concentration at FL and 5LED. The use of 0.1 µM TDZ at 5LED decreased the ratio of chlorophylls a + b to carotenoids and led to the highest accumulation of taxifolin and rutin, quercetin, hyperoside, and avicularin. Thus, it has been demonstrated that the application of combined action of LED and TDZ has great potential in terms of propagation efficiency, biomass accumulation, and taxifolin and rutin production in R. mucronulatum microshoots., Competing Interests: Conflict of InterestThe authors declare that they have no conflict of interest., (© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)