Effect of antioxidants and growth regulators on shoot organogenesis in the apical meristem culture of Fragaria × ananassa (Duchesne ex Weston) Duchesne ex Rozier

The initiation of strawberries into in vitro culture is known to be complicated by the inhibition of organogenesis by phenolic oxidation products. An important role in this process is given to the selection of growth regulators that increase meristematic cell activity and shoot proliferation at the stage of organogenesis induction. The present study aims to obtain a viable apical meristem culture of garden strawberry and to study the effect of different antioxidants (reduced glutathione (RG); a new preparation, i.e., a mechanical composite (MC) on the basis of biogenic silicon and green tea catechins and plant growth regulators (6-benzylaminopurine; thidiazuron) on the initiation of axillary shoot formation in strawberry meristem culture. Terminal buds containing an apical meristem and two leaf primordia isolated from the stolons of two garden strawberry cultivars (Sunny Meadow and Festival Chamomile) were used as primary explants for the initiation of strawberries into in vitro culture. It was found for the first time that the MC exhibits higher antioxidant activity as compared to reduced glutathione, reduces darkening of initial explants, as well as enhancing regeneration up to 13.0% at p ≤ 0.05. Furthermore, the best effect on the formation of microshoots per explant is observed toward the end of material introduction into in vitro culture when combining the MC with growth regulators in the culture medium. Here, the effect of strawberry cultivar on explant regeneration and the number of microshoots per explant are insignificant. It is concluded that the procedure for using the MC as an effective antioxidant during material initiation into the culture can be applied to the large-scale in vitro propagation of garden strawberries. Moreover, the technology for obtaining the MC from plant waste is environmentally friendly, which is a significant advantage for its use in in vitro technologies.

Primary morphogenesis of Sorbus Torminalis L. (Grantz) into in vitro culture

Nowadays in vitro methods, combined with ex situ and becoming an increasingly important means of preserving and maintaining the level of phytodiversity stability. Sorbus torminalis L. is a tree of the Rosaceae family, which grows on the territory of Ukraine, belongs to rare, valuable aboriginal species and is listed in the Red Book of Ukraine, with its protection status - endangered. The peculiarities of introduction of in vitro culture of perennial representatives of S. torminalis with the use of different types of explants, sterilizing substances, cultivation conditions and nutrient medium composition are presented in the paper. For in vitro culture of S. torminalis, annual shoots with apical and lateral buds 15-25 cm long are optimal. The influence of different sterilization options on the development of primary microshoots has been studied. For sterilization of artificially awakened and young shoots it is most effective to use 0.1% solution of AgNO3 (7 min) and 15% solution of H2O2 (10 min). The developed method of sterilization of S. torminalis explants provided 80-90% yield of aseptic plant material. It was found that the sterilization regime did not significantly affect the primary morphogenesis of explants and was uniform. The optimal components of nutrient media at the stage of introduction into vitro culture and primary morphogenesis of S. torminalis have been established. For the cultivation of different types of explants of S. torminalis, used WPM nutrient media with the addition of synthetic plant growth regulators 6-benzylaminopurine, thidiazuron, kinetin 0,5-1,5 mg mg∙l-1 and α-naphthylacetic acid 0,01-0,05 mg∙l-1 both alone and in combination with each other. In particular, for the regeneration of plants from the lateral and apical buds of explants is effective WPM medium with the addition of BAP 1,5 mg∙l-1 + 0,5 mg∙l-1 NAA and WPM + TDZ medium 0,5 mg∙l-1 with adding PVP 200 mg∙l-1. To induce the laying of additional buds and shoots on the explant from the apical meristems in the medium should be added 4,0 mg∙l-1 BAP + 0,01 mg∙l-1 NAA with the addition of PVP 200 mg∙l-1.

Cellular Processes in Human Ovarian Follicles Are Regulated by Expression Profile of New Gene Markers—Clinical Approach

In the growing ovarian follicle, the maturing oocyte is accompanied by cumulus (CCs) and granulosa (GCs) cells. Currently, there remain many unanswered questions about the epithelial origin of these cells. Global and targeted gene transcript levels were assessed on 1, 7, 15, 30 days of culture for CCs and GCs. Detailed analysis of the genes belonging to epithelial cell-associated ontological groups allowed us to assess a total of 168 genes expressed in CCs (97 genes) and GCs (71 genes) during long-term in vitro culture. Expression changes of the analyzed genes allowed the identification of the group of genes: TGFBR3, PTGS2, PRKX, AHI1, and IL11, whose expression decreased the most and the group of ANXA3, DKK1, CCND1, STC1, CAV1, and SFRP4 genes, whose expression significantly increased. These genes’ expression indicates CCs and GCs epithelialization processes and their epithelial origin. Expression change analysis of genes involved in epithelization processes in GCs and CCs during their in vitro culture made it possible to describe the most significantly altered of the 11 genes. Detailed analysis of gene expression in these two cell populations at different time intervals confirms their ovarian surface epithelial origin. Furthermore, some gene expression profiles appear to have tumorigenic properties, suggesting that granulosa cells may play a role in cancerogenesis.