In vitro Regeneration of Picralima nitida (Stapf). T. Durand & H. using Zygotic Embryo

Disinfected mature seed embryos of Picralima nitida, were cultured in MS medium supplemented with different concentrations and combinations of 2,4-D, BAP and NAA to determine an efficient protocol for in vitro propagation. Nine culture media made of combination of different components were used in a factorial design with three replications. Results showed up to 80 ± 4% disinfection rate with combination of triton x- 100 (0.2%) and sodium hypochlorite (30%). Embryo germination was highest on control medium. Rooting was higher (2±1 roots per embryo) after 4 weeks on control medium and on BAP supplemented medium at 0.8 μM while the longest root (1.5±0.5 cm) was observed on 2,4-D supplemented medium at 1.8 μM. Black soil was suitable for leaf formation (4 ± 2 leaves) and shoot elongation (2±1 cm) after 8 weeks in acclimatisation. These results show efficient disinfection, regeneration and acclimatisation of Picralima nitida. Plant Tissue Cult. & Biotech. 31(2): 143-151, 2021 (December)

Development of the prickly pear cactus Opuntia stricta (Haw.) Haw. (Cactaceae) in vitro in response to the replacement of potassium nitrate for a commercial kno 3 fertilizer

ABSTRACT: In micropropagation, potassium nitrate (KNO3), an ACS reagent grade chemical, used in the preparation of growing mediums is expensive and its procurement depends on bureaucratic procedures, as it is controlled by the Brazilian Army. This research to assessed the effect of replacing the ACS KNO3 for a commercially available fertilizer (KNO3- based) on the micropropagation of the prickly pear cactus (Opuntia stricta (Haw.) Haw. cv. Elephant Ear. Treatments used six different fertilizer concentrations (0, 0.5, 1, 1.5, 2 and 2.5 g L-1) and a control consisting of 1.9 g L-1 KNO3, as shown in the MS salts. The survival, size and number of sprouts and the value of fresh biomass were evaluated. After seedling acclimation, we assessed the survival, number of sprouts, length, and number of roots, racket formation, average fresh biomass mass, macronutrient absorption and morphological changes of the seedlings. Explants inoculated with fertilizers at concentrations of 0.0; 2.0 and 2.5 g L-¹ did not grow. The response of explants at concentrations of 0.5 and 1.5 g L-1 of the fertilizer were the same as those developed in a KNO3 medium, and at a concentration of 1.0 g L-1, in all variables, the means were higher than those of the control medium. Therefore, it showed the feasibility of using fertilizers in the in vitro cultivation of the prickly pear cactus, which may remove bureaucratic barriers and reduce product costs by 99.12%.

Polyscias filicifolia (Araliaceae) Hairy Roots with Antigenotoxic and Anti-Photogenotoxic Activity

Hairy root cultures are considered as a valuable source of bioactive phytoconstituents with expanding applicability for their production. In the present study, hairy root cultures of Polyscias filicifolia (Araliaceae), a traditional Southeast Asian medicinal plant, were established. The transformation with Agrobacterium rhizogenes ATCC 15834 allowed to obtain 15 root lines. The K-1 line, demonstrating the highest growth capabilities, was subjected to further investigations. To enhance the biosynthetic potential of hairy roots, methyl jasmonate elicitation approach was applied (MeJA; at different doses and exposure time), with subsequent transfer of elicited roots to control medium. This strategy resulted in chlorogenic acid production up to 1.59 mg/g dry weight. HPLC-PDA-ESI-MS analysis demonstrated variation in extracts composition and allowed to identify different caffeic and ferulic acid derivatives. Next, cytotoxic, antigenotoxic, and anti-photogenotoxic properties of hairy roots extracts were determined. None of the tested extracts were cytotoxic. In addition, they demonstrated significant antigenotoxic activity with the highest protective potential; up to 52% and 49% of inhibition of induction ratio (IR) induced by the 2-aminoanthracene was revealed for extracts derived from hairy roots elicited for 3 days with 50 µM MeJA and roots elicited for 7 days with 100 µM MeJA and then transferred for 30 days to control medium, respectively. These same extracts exhibited the highest anti-photogenotoxic potential, up to 36% of inhibition of chloropromazine-induced genotoxicity.

Dietary Behavior of Drosophila melanogaster Fed with Genetically-Modified Corn or Roundup®

With the rise in concern about GMOs and pesticides on human health, we have utilized Drosophila melanogaster as a model organism for understanding the effects of Roundup-Ready® GMO diets on health. We recorded dietary behavior during and after exposure to a medium containing GMO or non-GMO corn, Roundup® in organic corn medium, and sucrose with or without one of the two Roundup® formulations. No differences in behavior were observed when Drosophila were exposed to a medium containing Roundup-Ready® GMO or non-GMO corn. Drosophila can detect and refrain from eating sucrose containing one Roundup® formulation, Ready-to-Use, which contains pelargonic acid in addition to glyphosate as an active ingredient. Drosophila exhibited dose-dependent increased consumption of sucrose alone after exposure to a medium containing either Roundup® formulation. This may indicate that flies eating a medium with Roundup® eat less and were thus hungrier when then given sucrose solution; that a medium with Roundup® is more difficult to digest; or that a medium with Roundup® is less nutritious, as would be the case if nutritionally important microbes grew on control medium, but not one containing Roundup®.

On necessity for analytical solution of the Bloch equations for nuclear magnetic resonance signals at condition express control of liquid medium

The necessity of using express analysis methods to control medium condition is substantiated. It has been shown that the method of express control based on the phenomenon of nuclear magnetic resonance is one of the most preferable. It was found that to increase the information about the medium condition state obtained from the recorded NMR signal, it is necessary to use a mathematical model (based on analytical solutions of the Bloch equations). Two approaches are considered that are used to describe the NMR signal in a liquid medium. It is determined that in the classical approach in the system of Bloch equations it is possible to take into account the peculiarities of using radiotechnical methods of signal registration. The direction of the analytical solution of the Bloch equation is proposed. The experimental data are compared with the numerical solution.

Aloe vera increases collagen fibres in extracellular matrix and mRNA expression of peroxiredoxin-6 in bovine ovarian cortical tissues cultured in vitro

Summary In vitro culture of ovarian tissue containing primordial follicles is an important tool to study the initiation of follicular populations and to develop efficient culture systems to support in vitro follicle growth. Considering that in vitro culture favours oxidative stress, it is very important to supplement culture medium with antioxidant substances such as Aloe vera extract. This study aims to evaluate the effects of different concentrations of Aloe vera on the distribution of collagen fibres in the extracellular matrix, follicular activation, development and survival in bovine ovarian cortical tissues cultured in vitro, as well as on expression of mRNAs for antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), peroxiredoxin 6 (PRDX6) and glutathione peroxidase 1 (GPX1)]. To this end, ovarian cortical tissues were cultured for 6 days in α-MEM alone or supplemented with different concentrations of Aloe vera extract (1.0, 5.0, 10.0 or 50.0%). After culture, fragments were fixed and processed histologically to evaluate follicular morphology and activation, as well as the extracellular matrix by staining with picrosirius red. The levels of mRNA for SOD, CAT, PRDX6 and GPX1 in cultured ovarian tissues were evaluated by real-time polymerase chain reaction (PCR). Ovarian tissues cultured with 10.0 or 50.0% Aloe vera had higher percentages of collagen fibres than tissues cultured in control medium. A significant increase in developing follicles was observed in ovarian tissues cultured in α-MEM alone or supplemented with 10% Aloe vera when compared with fresh control or tissues cultured with 1.0% Aloe vera. Presence of Aloe vera did not influence the percentage of morphologically normal follicles when compared with control medium. Ovarian tissues cultured with 50.0% Aloe vera had higher percentages of morphologically normal follicles than those cultured with 10.0% Aloe vera. Furthermore, 10% Aloe vera significantly increased mRNA levels for PRDX6. In conclusion, 10.0% Aloe vera improves extracellular matrix distribution in cultured tissues and increases the expression of mRNA for PRDX6 after 6 days in vitro.

The Role of MAPK3/1 and AKT in the Acquisition of High Meiotic and Developmental Competence of Porcine Oocytes Cultured In Vitro in FLI Medium

The developmental potential of porcine oocytes cultured in vitro was remarkably enhanced in a medium containing FGF2, LIF and IGF1 (FLI) when compared to a medium supplemented with gonadotropins and EGF (control). We analyzed the molecular background of the enhanced oocyte quality by comparing the time course of MAPK3/1 and AKT activation, and the expression of genes controlled by these kinases in cumulus-oocyte complexes (COCs) cultured in FLI and the control medium. The pattern of MAPK3/1 activation in COCs was very similar in both media, except for a robust increase in MAPK3/1 phosphorylation during the first hour of culture in the FLI medium. The COCs cultured in the FLI medium exhibited significantly higher activity of AKT than in the control medium from the beginning up to 16 h of culture; afterwards a deregulation of AKT activity occurred in the FLI medium, which was not observed in the control medium. The expression of cumulus cell genes controlled by both kinases was also modulated in the FLI medium, and in particular the genes related to cumulus-expansion, signaling, apoptosis, antioxidants, cell-to-cell communication, proliferation, and translation were significantly overexpressed. Collectively, these data indicate that both MAPK3/1 and AKT are implicated in the enhanced quality of oocytes cultured in FLI medium.

Effects of Biochar and Biochar–Compost Mix on Growth, Performance and Physiological Responses of Potted Alpinia zerumbet

Container crop production has become increasingly popular over the last 50 years. A major component of container or potting media is peat. Peatlands are a natural carbon sink, and peat is a nonrenewable natural resource. Peat harvesting has become an important environmental issue. There is a growing effort to explore alternative organic materials to completely or partially replace peat as a medium component. Biochar is a carbon-rich product that has gained increasing interest as a component of growing media. In the present study, biochar was produced from rice straw. Peat/perlite/biochar (PPB; 40/30/30 v/v) and peat/perlite/biochar/vermicompost (PPBC; 30/30/35/5 v/v) were evaluated relative to a basal or control medium of peat/perlite (PP; 70:30 v/v). Alpinia (Alpinia zerumbet ‘Variegata Dwarf’) was used as a test plant. Amending biochar and biochar–compost mix increased the pH of the growing media. Hydrophysical properties including container capacity, bulk density, air space and total porosity were all within or near the standard ranges for soilless growing media. Chlorophyll a and b contents of A. zerumbet plants grown in PPB medium were reduced by more than 20% and 28%, respectively, compared to those grown in PP or PPBC media. The net photosynthetic rate of PPB-grown plants was more than 28% lower than those grown in PP and PPBC media. As a result, shoot and root dry weights of plants produced in PPB medium were more than 42% and 22% less, respectively, than those grown in PP and PPBC media. Although visual quality of PPB-grown plants was lower, they still exhibited marketable quality, which was largely due to the fact that their side shoots, leaf numbers, leaf areas, leaf thickness, and shoot diameters were comparable to those produced in PP and PPBC media. The present study showed that in a peat/perlite basal medium, substitution of peat by biochar derived from rice straw at 30% affected the growth of A. zerumbet plants, mainly in dry matter accumulation, but the plants were still marketable. On the other hand, plants grown in the same basal medium with peat replaced by the biochar at 35% plus an amendment of compost at 5% were comparable to those grown in the control medium. As the value of ornamental plants depends on their aesthetic appearance, a potting medium comprised of peat/perlite/biochar/vermicompost at 30/30/35/5 by volume is recommended for the production of A. zerumbet plants. The substitution of peat at 35% suggests that peat use can be reduced in the formulation of potting media, thus contributing to the conservation of peatlands.

A blend of 3 mushrooms dose-dependently increases butyrate production by the gut microbiota

The gut microbiota has been indicated to play a crucial role in health and disease. Apart from changes in composition between healthy individuals and those with a disease or disorder, it has become clear that also microbial activity is important for health. For instance, butyrate has been proven to be beneficial for health, because, amongst others, it is a substrate for the colonocytes, and modulates the host’s immune system and metabolism. Here, we studied the effect of a blend of three mushrooms (Ganoderma lucidum GL AM P-38, Grifola frondosa GF AM P36 and Pleurotus ostreatus PO AM-GP37)) on gut microbiota composition and activity in a validated, dynamic, computer-controlled in vitro model of the colon (TIM-2). Predigested mushroom blend at three doses (0.5, 1.0 and 1.5 g/day of ingested mushroom blend) was fed to a pooled microbiota of healthy adults for 72 h, and samples were taken every day for microbiota composition (sequencing of amplicons of the V3-V4 region of the 16S rRNA gene) and activity (short-chain fatty acid (SCFA) production). The butyrate producing genera Lachnospiraceae UCG-004, Lachnoclostridium, Ruminococcaceae UCG-002 and Ruminococcaceae NK4A214-group are all dose-dependently increased when the mushroom blend was fed. Entirely in line with the increase of these butyrate-producers, the cumulative amount of butyrate also dose-dependently increased, to roughly twice the amount compared to the control (medium without mushroom blend) on the high-dose mushroom blend. Butyrate proportionally made up 53.1% of the total SCFA upon feeding the high-dose mushroom blend, compared to 27% on the control medium. In conclusion, the (polysaccharides in the) mushroom blend led to substantial increase in butyrate by the gut microbiota. These results warrant future mechanistic research on the mushroom blend, as butyrate is considered to be one of the microbial metabolites that contributes to health, by increasing barrier function and modulating inflammation.

Effect of adding anethole to the cryopreservation medium (powdered coconut water) on morphology, kinetics, and oxidative stress of buck sperm

The quality of post-thawing goat sperm is critical to the success of artificial insemination protocols and may be influenced by extenders, cryoprotectants, and antioxidant substances. Therefore, the objective of this study was to evaluate the effects of the antioxidant anethole on goat sperm diluted in preservation medium based on powdered coconut water (ACP-101c) and frozen. For that, each ejaculate was submitted to the following treatments: ACP-101c (control); control plus supplementation with 30, 300, or 2000 μg/mL anethole. The samples were thawed and evaluated for morphology, kinetics, membrane integrity, and reactive oxygen species (ROS). The addition of anethole increased morphological abnormalities (P < 0.05), however, it did not affect sperm kinetics. Flow cytometry analysis showed that sperm cells cryopreserved with 300 μg/mL anethole had lower acrosome integrity than those cryopreserved in other treatments. Evaluation of oxidative stress revealed that cells stored in the presence of 2000 μg/mL anethole had small amounts of ROS when compared to those preserved in the control medium alone or supplemented with 300 μg/mL anethole (P < 0.05). After cryopreservation of sperm with 2000 μg/mL anethole, the highest percentage of viable sperm without ROS was observed (P < 0.05). In conclusion, despite reducing ROS levels, the supplementation of anethole in ACP-101c did not affect sperm kinetics or membrane integrity post-thawing, however, it did cause morphological damage to sperm.