Bursera fagaroides popularly used in México possess bioactive lignans. These compounds are low in the bark, and its extraction endangers the life of the trees. The aim of the present investigation was to search for alternative sources of cytotoxic compounds from B. fagaroides as leaves and in vitro callus cultures. The friable callus of B. fagaroides were established using a combination of phytoregulators: 4 mgL-1 of 2,4-dichlorophenoxyacetic acid (2,4-D), 1 mgL-1 Naphthaleneacetic Acid (NAA) and 1 mgL-1 Zeatin. The maximum cell growth was at day 28 with a specific growth rate μ= 0.059 days-1 and duplication time td=11.8 days. Scopoletin with a concentration of 10.7 µg/g dry weight was the main compound inducible as a phytoalexin by the addition of high concentrations of 2,4-D, as well as by the absence of nutrients in the culture medium. The com-pounds γ-sitosterol, and stigmasterol were also present. Open column chromatography was used to separate and identify: yatein, acetyl podophyllotoxin and 7'8'-dehydropodophyllotoxin in the leaves of the wild plant. Cytotoxic activity on four cancer cell lines was tested, being PC-3 prostate carcinoma (IC50 of 12.6±4.6) the most sensitive to the wild-type plant extract and HeLa cervical carcinoma (IC50 of 72±5) to the callus culture extract.

Plant stem cell cultures have so far been established in only a few plant species using cambial meristematic cells. The presence of stem cells or stem cell-like cells in other organs and tissues of the plant body, as well as the possibility of de novo generation of meristematic cells from differentiated cells, allows to consider the establishment of stem cell cultures in a broader range of species. Therefore, this study aimed to establish a stem cell culture of the medicinal plant Calendula officinalis L. Callus tissues were induced from explants taken from leaves and roots. Appropriate combinations of plant growth regulators and light parameters for in vitro cultivation were selected. Already at this stage, stem and dedifferentiated cells could be identified, and cell suspension cultures were established from specific parts of the formed callus. Cell suspension cultures with a high proportion of stem cells originating from roots (92–93%) or leaves (72–73%) were developed. The amount of stem cells derived from the roots in the cell suspension reached a plateau at a level of 5.60–5.72105.

The evaluation of the possibility of obtaining dihaploids by the method of anther culture in vitro to accelerate selection for resistance to prolonged flooding of seeded rice with water was carried out. The object of research is rice hybrids F2 of the Rice Breeding and Seed Production Laboratory of the “Donskoy” ASC, obtained as a result of interbreeding the best economically valuable varieties with samples bearing genes of resistance to prolonged flooding with water. Basic nutrient medium with an optimal composition of nutrition elements and growth hormones stimulating callo- and morphogenesis were used. For the induction of callus formation, 12604 anthers were planted, according to 26 hybrid combinations represented by 68 plants, as a result, 716 neoplasms were obtained, including 586 non–morphogenic callus, 130 regenerating plants. Cultivation of anthers revealed large genotypic differences in the samples. According to the responsiveness to neoplasms, 60% of the panicles showed a positive result, the rest did not give callus. The most responsive to the formation of callus were hybrid combinations: 5009/2 – 84 pcs., 5010/2 – 94 pcs., 4565/3 – 85 pcs., 4641/2 – 69 pcs. They also showed the ability to morphogenesis. Androgenic plants were obtained by 14 hybrid combinations, their share was 1.03% of the total number of inoculated anthers. 30 green regenerative lines from four rice hybrids were obtained, differing in visual morphological assessment: 5009/2 - 5 pcs., 5010/2 – 5 pcs., 4565/3 – 2 pcs., 4641/2 – 18 pcs. The isolated lines are characterized by good responsiveness in the culture of anthers in vitro, carry genes for resistance to prolonged flooding, and can be used in rice breeding programs using DG technologies.

Thai basil is a renowned medicinal plant and a rich source of bioactive antioxidant compounds having several health benefits, with actions to prevent of cancer, diabetes and cardiovascular disease. Plant cell and tissue culture technologies can be routinely established as an important, sustainable and low-cost biomass source for the production of high-value phytochemicals. The current study aimed at developing an effective protocol for the production of Thai basil leaf derived callus cultures with sustainable and high production of biomass and antioxidants as an alternative of leaves production. MS basal medium with various concentrations of plant growth regulators (PGRs) compatible with nutraceutical applications (i.e., gibberellic acid (GA3) and 6-benzylaminopurine (BAP) either alone or in combination with naphthalene acetic acid (NAA)) were evaluated. Among all tested PGRs, the combination BAP:NAA (5 mg/L:1 mg/L) yield maximum biomass accumulation (fresh weight (FW): 190 g/L and dry weight (DW): 13.05 g/L) as well as enhanced phenolic (346.08 mg/L) production. HPLC quantification analysis indicated high productions of chicoric acid (35.77 mg/g DW) and rosmarinic acid (7.35 mg/g DW) under optimized callus culture conditions. Antioxidant potential was assessed using both in vitro cell free and in vivo cellular antioxidant assays. Maximum in vitro antioxidant activity DPPH (93.2 % of radical scavenging activity) and ABTS (1322 µM Trolox equivalent antioxidant capacity) was also observed for the extracts from callus cultures grown on optimal conditions. In vivo cellular antioxidant activity assay confirmed the effective protection against oxidative stress of the corresponding extract by the maximum inhibition of ROS and RNS production. Compared to commercial leaves, callus extracts showed higher production of chicoric acid and rosmarinic acid associated with higher antioxidant capacity. In addition, this biological system also has a large capacity for continuous biomass production, thus demonstrating its high potential for possible nutraceutical applications.

Callus induction of F. moluccana (sengon) was still an obstacle to indirect organogenesis regeneration. The purpose of the study was to determine the callus induction formation from root fragments of F. moluccana plantlets. Primary explants (fragments of roots) were cultivated on MS induction basal media and three concentration combination of PGRs (BAP and NAA): 0.5 ml/l BAP; 0,1 ml/l NAA (single PGR); and combination of 0.5 ml/l BAP + 0.1 ml/l NAA (double PGR). When roots were used as explants, high formation rates of callus (more than 70%) were obtained. Highest formation rates of callus were in NAA added at all clones (12 clones), then BAP added (7 clones) and BAP + NAA added (5 clones). The results indicated that BAP and NAA concentrations used in the media were influence the producing callus and affect the amount of callus produced from roots of sengon. The addition of NAA also gives higher callus proliferation results than the addition of BAP or the addition of a combination of the two hormones. The results indicated that BAP and NAA concentrations used in the media were influence the producing callus and affect the amount of callus produced from roots of sengon.

Wounding and exogenous auxin are needed to induce adventitious roots in Chestnut microshoots. However, the specific inductive role of wounding has not been characterized in this species. In the present work two main goals were established: First, we prompted to optimize exogenous auxin treatments to improve overall health status of the shoots at the end of the rooting cycle. Second, we developed a time-series transcriptomic analysis to compare gene expression in response to wounding alone and wounding plus auxin, focusing in the early events within the first days after treatments. Results suggest that the expression of many genes involved in the rooting process is under direct or indirect control of both stimuli. However, specific levels of expression of relevant genes are only attained when both treatments are applied simultaneously, leading to the successful development of roots. In this sense, we have identified four transcription factors upregulated by auxin (CsLBD16, CsERF113, Cs22D, CsIAA6), with some of them being also induced by wounding. The highest expression levels of these genes occurred when wounding and auxin treatments were applied simultaneously, correlating with the rooting response of the shoots. Results in this work clarify the genetic nature of the wounding response in chestnut, its relation to adventitious rooting and might be helpful in the development of more specific protocols for the vegetative propagation of this species.

Exosomes are membrane-bound, biologically active nanovesicles of size 30-200 nm produced by many cell types, such as both mammalian and plant cells. They are wrapped in a phospholipid bilayer and play a significant role in intercellular communications. The ease in their isolation and the ability of plant-derived exosome-like nanovesicles (PDEVs) to efficiently deliver bioactive constituents into mammalian cells have made them popular in contemporary research. PDEVs share many characteristics with mammalian EVs (MEVs), including shape, size, surface charge, and consist of bioactive molecules like lipids, proteins, nucleic acids, and tiny metabolites. However, the chemical composition profile of PDEVs and their biogenesis mechanism differ significantly from those of MEVs. They have been widely explored as potential therapeutic agents and are considered as good alternatives to act as a carrier for drug delivery. The present work elucidates the isolation of exosome-like-nanovesicles (henceforth exosomes) from the culture supernatants of an in vitro cultured callus tissue derived from a bone healing plant known for its osteogenic activity, i.e., Cissus quadrangularis. The physical and biological properties of exosomes were successfully studied using different characterization techniques. To assess their therapeutic potential, we studied the internalisation of calcein-AM labeled exosomes by human derived mesenchymal stromal cells (hMSCs). Additionally, we evaluated the potential of exosomes in the migration of cells in a cell scratch assay with hMSCs and their effect on amelioration of oxidative stress was investigated on preosteoblast MC3T3-E1 cells that were pre-treated with these exosomes. Furthermore, we investigated their proliferation and differentiation to osteoblasts like cells with the help of resazurin assay and alkaline phosphatase assay (ALP). The obtained results provide a primary justification for the use of Cissus quadrangularis-derived exosomes as a nanocarrier for drug molecules for various therapeutic bone applications.

Plant cell and tissue culture has been a crucial component of efforts to enhance crops and advance plant biotechnology ever since the beginning of the XX century when Hildebrand discovered the theory of totipotency – a somatic plant cell still has the capacity to develop into every cell that makes up a whole plant. Further standardization of the techniques for use in crop improvement has resulted from the original interest with the creation of the numerous plant tissue culture techniques through the various advancement. The most important approach used in plant tissue culture is the cultivation of differentiated tissue that results in the development of an undifferentiated mass of parenchymatous cells and their subsequent differentiation by organogenesis or somatic embryogenesis. This has been playing a significant role in the creation of virus-free seedlings and fast clonal multiplication, and it is also a crucial component in transgenic' regeneration. Additionally, cell cultures with a variety of mutations emerging in it offers chances to enhance crops. For the purpose of identifying homozygous recombinants in a single generation, haploid cell culture offers an additional benefit. Wide hybridization and the embryo rescue procedure have helped to produce haploid embryos and homozygous lines of doubled haploids which are difficult and time consuming to produce using traditional methods. Developed haploid embryos from the male gametophyte, carried out in cultures of immature anthers or isolated microspores, are referred to as androgenesis and could be the most efficient way to create lines of doubled haploids. The cell cultures also offer the chance to extract a variety of beneficial biochemicals. Induction of mutations in cultured cells and tissue in vitro to produce beneficial cultivars and manipulation of the centromere-specific histone CENH3 also become possible in recent years. Compared to traditional methods, in vitro cultures offer a number of both scientific and practical advantageous for scientists and plant breeders. However, the usefulness of individual methods should be examined by analyzing their advantages and disadvantages. When it comes to oat (Avena sativa L.) the efficient method of plant regeneration is still missing compared to the most common cereals, possibly because this cereal is known as recalcitrant for in vitro culture. An effort has been made to provide a succinct overview of the various in vitro techniques utilized or potentially involved in breeding of oat (Avena sativa L.) in this review. The present work aims to summarize the crucial methods of A. sativa cultivation under tissue culture conditions with a focus on the progress that has been made in biotechnological techniques which are used in breeding of this species.

Somatic embryogenesis-mediated plant regeneration is essential for genetic manipulation of agronomically important traits in upland cotton. Genotype specific recalcitrance to regeneration is a primary challenge in deploying genome editing and incorporating useful transgenes into elite cotton germplasm. In this study, transcriptomes of a semi-recalcitrant cotton (Gossypium hirsutum L.) genotype ‘Coker312’ were analyzed at two critical stages of Somatic Embryogenesis that includes non-embryogenic callus (NEC) and embryogenic callus (EC) cells, and the results compared to a non-recalcitrant genotype ‘Jin668’. We discovered of 305 differentially expressed genes in Coker312, whereas, in Jin668, about 6-fold more genes (2,155) were differentially expressed. A total of 154 differentially expressed genes were common between the two genotypes. Gene enrichment analysis of upregulated genes identified functional categories such as lipid transport, embryo development, regulation of transcription, sugar transport, vitamin biosynthesis, among others. In Coker312 EC cells, five major transcription factors were highly upregulated: LEAFY COTYLEDON 1 (LEC1), WUS-related homeobox 5 (WOX5), ABSCISIC ACID INSENSITIVE3 (ABI3), FUSCA3 (FUS3), and WRKY2. In Jin668, LEC1, BABY BOOM (BBM), FUS3, and AGAMOUS-LIKE15 (AGL15) were highly expressed in EC cells. We also found that gene expression of these embryogenesis genes was typically higher in Jin668 when compared to Coker312. We conclude that significant differences in expression of the above genes between Coker312 and Jin668 may be a critical factor affecting the regenerative ability of these genotypes.

Conventional fluorescent lamps used in tissue culture are costly light sources showing excessive wavelength emission-bandwidth that must be replaced by alternative, less costly and much lower power-consuming energy sources. The use of Light-Emitting Diodes (LEDs) is the best option due to their potential role as elicitors of secondary metabolite production in many plant models. Gynura procumbens(G. procumbens) is widely used for treating various diseases. Here, leaf explants were cultivated in MS medium supplemented with 0.5 mg/L of NAA and 2.0 mg/L of BAP for 30 days under white, blue, and red LEDs. Secondary metabolites were analyzed by HPLC and LC-MS. Blue LEDs elicited the highest antioxidant activity, total flavonoid, and phenolic content. Furthermore, the content of cyanidin-monoglucosides increased significantly increased under blue light.

The viability, productivity and survival of higher plants under the adverse factors influence are largely determined by the functional activity of the antioxidant system. The aim of our work was to investigate changes in formation of high-molecular (superoxide dismutase and peroxidase) and low-molecular (phenolics, including flavanols and proanthocyanidins) antioxidants in callus culture of Camellia sinensis under influence of phenolic precursors (L-phenylalanine – 3 mM, trans-cinnamic acid – 1 mM, naringenin – 0.5 mM). According to the data obtained, precursors effect on tea callus cultures did not lead to significant increasing of superoxide dismutase and peroxidase activity in most cases. However, it led to the increased accumulation of the total phenolics content, as well as flavanols and proanthocyanidins contents. For C. sinensis callus cultures, the most promising regulator of phenolic compounds was L-phenylalanine, in the presence of which its content increased almost twice. Thus, the exogenous effect of various precursors is possible to use for the targeted regulation of the certain phenolics classes accumulation in plant cells.