Providing inoculum for Didymella bryoniae studies: the effect of light spectrum and storing at low temperature

The in vitro sporulation of Didymella bryoniae is of great importance for studies that require pure inoculum and in large quantities. Thus, the objectives of this study were to identify the best condition for D. bryoniae sporulation combining different light spectra (UV-A or UV-B light, white light, and continuous dark), with distinct culture media (PDA, V8, ML, and PDAB) and, to evaluate fungus’ survivability stored at -20°C over time. The fungus samples were only able to sporulate when subjected to the UV-B light treatment, regardless of the culture medium. The highest appearance of spores conidium type was observed in the PDAB medium, and the lowest production occurred in the ML medium. Reproductive structures, such as perithecia and pycnidia, were observed in all culture media. However, there was considerable variation in the amount of each structure between the different culture media. The ML and V8 media showed a greater number of perithecia and the PDA and PDAB media presented a greater proportion of pycnidia compared to perithecia. The storage duration at -20°C did not affect mycelial growth or mycelial growth rate. In conclusion, the UV-B light is essential for D. bryoniae in vitro sporulation. Moreover, the culture medium composition influences the type of fungal structure produced, as well as spores’ size and quantity. Freezing at -20°C is an efficient technique that can be used to store D. bryoniae for at least five months without loss of viability.

Morphogenesis of Anthers Isolated From Cabbage (Brassica Oleracea L.) In Vitro

This study aimed to optimize the steps of obtaining regenerated cabbage plants by direct embryogenesis from isolated anthers and ovaries. Stepwise pretreatment of inflorescences was usedfor the studied hybrids and inbred lines. First, the inflorescences were placed in water and kept at a temperature of +4-6∘C for one day without the use of biologically active substances. Then the inflorescences were placed in a solution of the drug Dropp (10 mg/l) and cultivated for two days. After that, the anthers and ovaries were isolated from the flower buds and cultured on the MS culture medium at a temperature of + 32∘C for one day. The cultivation of the isolated explants on a nutrient medium (containing 0.01 mg/lof Dropp, 1.0 mg/lof NAA, 500 mg/lof asparagine, 100 mg/l of tyrosine, and 10 g/l of sucrose)led to an increase in their morphogenetic potential in the culture of anthers and ovaries (by 3.42% and 5.54%, respectively).A cytological method was usedto demonstrate the haploid nature of the regenerating plants. The number of chromosomes in the root meristem andleaves, and the chloroplasts in the closing cells of the stomatawere calculated. Keywords: cabbage, culture in vitro, regenerated plants, anthers, ovaries, reproductive organs

Effect of conditioned medium of umbilical cord-derived mesenchymal stem cells as a culture medium for human granulosa cells: An experimental study

Background: The umbilical cord-derived mesenchymal stem cells conditioned medium (UC-MSCs-CM) produces secretomes with anti-apoptotic properties, and has the potential to prevent apoptosis of granulosa cells (GC) during controlled ovarian hyperstimulation. Objective: To observe the effect of UC-MSCs-CM on the interaction between pro- and anti-apoptotic proteins and the influence of growth differentiation factor 9 (GDF9) production in GC. Materials and Methods: UC-MSCs-CM was collected from umbilical cord stem cell culture on passage 4. GC from 23 women who underwent in vitro fertilization were cultured and exposed to UC-MSCs-CM for 24 hr. Then RNA of the GC was extracted and the mRNA expression of BCL-2 associated X (BAX), survivin and GDF9 were analysed using quantitative real-time PCR. The spent culture media of the GC were collected for measurement of insulin growth factor 1 using ELISA. Results: The expression of BAX was significantly different after UC-MSCs-CM exposure (4.09E-7 vs. 3.74E-7, p = 0.02). No significant changes occurred in survivin, BAX/survivin ratio, and GDF9 expression after UC-MSCs-CM exposure (p > 0.05). The IGF-1 level of the CM was significantly higher after the CM was used as a culture medium for GC (2.28 vs. 3.07 ± 1.72, p ≤ 0.001). A significant positive correlation was found between survivin and GDF9 (r = 0.966, p ≤ 0.001). Conclusion: IGF-1 produced by UC-MSCs-CM can work in paracrine fashion through the IGF receptor, which can inhibit BAX and maintain GDF9 production. Moreover, under the influence of UC-MSCs-CM, GC are also capable of producing IGF-1, which can impact GC through autocrine processes. Key words: Conditioned medium, BAX, Survivin, GDF9, IGF-1.

Photo-induced antifungal activity of chitosan composite film solution with Nano TiO2 and Nano Ag

This study investigated the ultraviolet (UV) light-induced effect of chitosan-titanium dioxide-silver (CTS-TiO2-Ag) nanocomposite film solution against Penicillium steckii ( ( P. steckii ) , as well as the underlying the physiological mechanism. The results indicated that the longer the UV exposure time, the better the pathogenic inhibition effect. After UV photoinduced treatment for 120 min, the colony diameter of P. steckii was the smallest at 4.85 mm. However, when this process is followed by an 8-h storage period, the conductivity of the P. steckii culture medium reached its highest level at 713 μs/cm. After a 120 h growth period in the same conditions, the lesion diameters and pathogenicity of the mangoes reached 12.61 mm and 41.67%, respectively. Since the cell membrane was severely disrupted, its permeability increased, causing serious intracellular protein and nucleic acid material extravasation. Furthermore, the malondialdehyde (MDA) , catalase (CAT) and superoxide dismutase (SOD) in the P. steckii reached maximum levels after 8 h of incubation, at 2.1106 μmol/mL, 44.06 U/mL, and 24.67 U/mL respectively. These results indicated significant P. steckii inhibition via the UV light induction of the CTS-TiO 2 -Ag composite film solution.

Russian and Foreign Cultivars of Honeysuckle (Lonicera edulis Turcz.): cultivation studies in vitro

Introduction. The demand for honeysuckle berries and planting material is growing. Clonal micropropagation is the most effective method for industrial plantations. The research objective was to study the effect of cytokinins and auxins on Russian and Canadian honeysuckle microshoots and roots. Study objects and methods. The study featured regenerated honeysuckle (Lonicera edulis Turcz.) of three Russian cultivars (Bakcharsky Velikan, Doch Velikana, Yugana) and two Canadian cultivars (Boreal Beauty, Boreal Beast). The experiment focused on the effect of sterilizing agents and sterilization time on the viability of honeysuckle explants at the stage of culture introduction in vitro. The effect of the growth regulator Cytodef in the QL nutrient medium on organogenesis was studied at the stage of micropropagation proper, the effect of auxin IBA on plant root formation – at the stage of rooting in vitro. Results and discussion. The greatest viability of honeysuckle explants (80–94%) was registered in the samples affected by Lizoformin 3000 (5%) and silver nitrate (0.2%) as sterilizing agents with a sterilization time of 10 min at the stage of in vitro culture introduction. The biggest quantity (8.8 pcs.) and total length (40.1 cm) of microshoots were observed when the content of cytokinin Cytodef in the culture medium QL was 0.3 mg/L at the stage micropropagation proper. The Boreal Beast cultivar had the largest total length of shoots (29.0 cm). The biggest quantity (5.5 pcs.) and total length (30.8 cm) of roots resulted from 0.5 mg/L of auxin IBA at the stage of rooting in vitro. Coconut substrate produced the highest survival rate (92–99%) at the stage of adaptation to non-sterile conditions in vivo, with the greatest number of leaves (8.1–10.2 pcs.) observed in Canadian cultivars. Conclusion. Cytodef and IBA proved to be effective growth-regulating substances for microplants of Russian and Canadian honeysuckle cultivars in vitro, which makes them promising for berry plantations.

Yeast Protein as an Easily Accessible Food Source

In recent years, the awareness and willingness of consumers to consume healthy food has grown significantly. In order to meet these needs, scientists are looking for innovative methods of food production, which is a source of easily digestible protein with a balanced amino acid composition. Yeast protein biomass (single cell protein, SPC) is a bioavailable product which is obtained when primarily using as a culture medium inexpensive various waste substrates including agricultural and industrial wastes. With the growing population, yeast protein seems to be an attractive alternative to traditional protein sources such as plants and meat. Moreover, yeast protein biomass also contains trace minerals and vitamins including B-group. Thus, using yeast in the production of protein provides both valuable nutrients and enhances purification of wastes. In conclusion, nutritional yeast protein biomass may be the best option for human and animal nutrition with a low environmental footprint. The rapidly evolving SCP production technology and discoveries from the world of biotechnology can make a huge difference in the future for the key improvement of hunger problems and the possibility of improving world food security. On the market of growing demand for cheap and environmentally clean SPC protein with practically unlimited scale of production, it may soon become one of the ingredients of our food. The review article presents the possibilities of protein production by yeast groups with the use of various substrates as well as the safety of yeast protein used as food.

Influence of N-Arachidonoyl Dopamine and N-Docosahexaenoyl Dopamine on the Expression of Neurotrophic Factors in Neuronal Differentiated Cultures of Human Induced Pluripotent Stem Cells under Conditions of Oxidative Stress

Oxidative stress (OS) is implicated in the pathogenesis of several neurodegenerative diseases. We have previously shown that N-acyl dopamines (N-ADA and N-DDA) protect the neural cells of healthy donors and patients with Parkinson’s disease from OS. In this study, we assessed the effects of N-acyl dopamines on the expression of neurotrophic factors in human-induced pluripotent stem cell-derived neuronal cultures enriched with dopaminergic neurons under conditions of OS induced by hydrogen peroxide. We showed that hydrogen peroxide treatment increased BDNF but not GDNF mRNA levels, while it did not affect the secretion of corresponding proteins into the culture medium of these cells. Application of N-acyl dopamines promoted BDNF release into the culture medium. Under conditions of OS, N-DDA also increased TRKB, TRKC and RET mRNA levels. Furthermore, N-acyl dopamines prevented cell death 24 h after OS induction and promoted the expression of antioxidant enzymes GPX1, GPX7, SOD1, SOD2 and CAT, as well as reduced the BAX/BCL2 mRNA ratio. These findings indicate that stimulation of the expression of neurotrophic factors and their receptors may underlie the neuroprotective effects of N-acyl dopamines in human neurons.