BA and TDZ in the Morphogenesis in Vitro of Passiflora Setacea

Passiflora setacea DC (Passifloraceae) is considered an important species in the genetic breeding of passion fruit. However, its use is limited due to low seed germination. This paper aimed to study the effect of cytokinins 6-benzyladenine (BA) and thidiazuron (TDZ) on the in vitro morphogenesis of P. setacea using three explants: hypocotyl, nodal segment, and root segment. The explants were induced to morphogenesis in MS medium modified and with different concentrations of BA and TDZ. After 55 days, the percentage of calluses and shoots were evaluated, and anatomical characterization was performed. The three explants used in the in vitro morphogenesis of P. setacea showed callus and shoots formation, but in greater numbers in the nodal segments treated with BA. TDZ isolated affected equal to or less than BA on callus and shoots formation for the three explants. Direct and indirect organogenesis was observed in the three types of explants. From the results obtained for plant regeneration via in vitro morphogenesis of P. setacea, it is recommended to use a nodal segment in MSM medium and supplemented with 2.22 μmol L-1 of BA.

WHEAT ANDROCLINIC CALLI: DATA OF SCANNING ELECTRONIC MICROSCOPY

The processes of formation different types of calli, as well as the morphogenesis pathways in morphogenic calli, were studied by scanning electron microscopy (SEM) during anther culture in vitro in hybrid line Fotos of spring soft wheat. The microspore haploid origin of calli has been proven. The morphological status of the obtained calli was determined. It was shown that morphogenic callus consists of small densely packed meristematic cells covered with extracellular substance. This type of calli was obtained using a variant of the Potato II induction culture medium, added by 1.0 mg/l synthetic auxin 2,4-D. Nonmorphogenic callus consists of large, elongated, loosely located cells with a smooth surface. This type of calli was obtained using a variant of the Potato II culture medium, added by 2.0 mg/l 2,4-D. It was found that the introduction of various IAA concentrations into the Blaydes nutrient medium for regeneration in morphogenic calli implements the following pathways of morphogenesis in vitro: embryoidogenesis (without IAA addition), gemmorhizogenesis (0.5 mg/l), and rhizogenesis (1.5 mg/l). Revealed degenerative changes in cells of nonmorphogenic calli. The fundamental possibility of regulating of the morphogenesis pathways of in vitro of morphogenic calli in the direction necessary for research in biotechnological research has been confirmed.

FAK Shutdown: Consequences on Epithelial Morphogenesis and Biomarker Expression Involving an Innovative Biomaterial for Tissue Regeneration

By employing an innovative biohybrid membrane, the present study aimed at elucidating the mechanistic role of the focal adhesion kinase (FAK) in epithelial morphogenesis in vitro over 4, 7, and 10 days. The consequences of siRNA-mediated FAK knockdown on epithelial morphogenesis were monitored by quantifying cell layers and detecting the expression of biomarkers of epithelial differentiation and homeostasis. Histologic examination of FAK-depleted samples showed a significant increase in cell layers resembling epithelial hyperplasia. Semiquantitative fluorescence imaging (SQFI) revealed tissue homeostatic disturbances by significantly increased involucrin expression over time, persistence of yes-associated protein (YAP) and an increase of keratin (K) 1 at day 4. The dysbalanced involucrin pattern was underscored by ROCK-IISer1366 activity at day 7 and 10. SQFI data were confirmed by quantitative PCR and Western blot analysis, thereby corroborating the FAK shutdown-related expression changes. The artificial FAK shutdown was also associated with a significantly higher expression of filaggrin at day 10, sustained keratinocyte proliferation, and the dysregulated expression of K19 and vimentin. These siRNA-induced consequences indicate the mechanistic role of FAK in epithelial morphogenesis by simultaneously considering prospective biomaterial-based epithelial regenerative approaches.

Preservation of representatives the genus Drosera L. using biotechnological methods

The peculiarities of obtaining planting material of rare representatives Drosera spatulate L. and Drosera aliciae L. using microclonal propagation in order to preserve and cultivate them in ex vitro conditions were studied. The method of sterilization of D. spatulate and D. aliciae explants with 80-90% obtaining aseptic material has been developed. The influence of different sterilization options on the development of microshoots has been studied. The best mode of sterilization is 0,1% solution of AgNO3 and 12.5% solution of H2O2. The features of organogenesis and regeneration of the whole organism from cultivated tissues and organs of Drosera L. was investigated. The effect of exogenous growth regulators at different stages of plant morphogenesis in vitro is shown. Improved conditions of rhizogenesis in vitro. It was found experimentally that MS nutrient media with the addition of 2 g∙l-1 PVP is optimal at the stage of introduction into culture in vitro D. spatulate and D. aliciae. The regeneration of microshoots of D. spatulate and D. aliciae depending on the type of explant and the composition of nutrient media was studied. Morphogenesis was most effective on nutrient media with the addition of 0.25 mg∙l-1 kinetin and on the hormone free MS media. Such cultivation conditions provided 100% regeneration of plants with a reproduction rate of 1:8. Studying the effect of cytokinins on the microclonal reproduction of D. spatulate and D. aliciae, it was found that the development and induction of multiple shoot formation in vitro is best performed on hormone free MS media. To induce the formation of the root system, it is necessary to add into MS nutrient media 0.5 mg∙l-1 IBA. According to the results of the research, a method of microclonal propagation was developed by cutting stem culture, which made it possible to obtain genetically stable, disease-free regenerating plants of D. spatulate and D. aliciae with an optimally formed root system and vegetative mass. The obtained homogeneous planting material can be used in floriculture, creation of terrariums, for pharmacological purposes and for the purpose of introduction. Keywords: Drosera L., microclonal reproduction, morphogenes.

Morphogenesis in vitro and peroxidase activity in barley cv. Steptoe and its ABA-deficient mutant AZ34: effects of inhibitors of ABA synthesis and auxin transport

The relationship between the effect of ABA on morphogenesis in vitro and auxin transport, as well as the role of peroxidases in the action of ABA on morphogenesis in vitro in the ABA-deficient barley mutant AZ34 and its parent form cv. Steptoe was studied.