Micropropagation of Feverfew (Tanacetum parthenium) and Quantification of Parthenolide Content in Its Micropropagated and Conventionally Grown Plants

Feverfew (Tanacetum parthenium) is a well-known multi-functional plant with anti-inflammatory, cardiotonic, antiangiogenic, and anticancer effects. The therapeutic value of this plant is due to its phytochemical constitutes, especially parthenolide. Tissue culture techniques have been applied to improve the bioactive components of many herbal plants. Hence, this study, was carried out to establish a protocol for micropropagation of the feverfew plant and to quantify parthenolide content in its micropropagated and conventionally grown plants. To establish an aseptic culture, different concentrations of sodium hypochlorite (NaOCl) were investigated for seed surface sterilization. Besides, the effects of plant growth regulators (PGRs) on the callus induction, shoot organogenesis from callus and in vitro rooting were evaluated. Additionally, the parthenolide yield of the micropropagated and conventionally grown plants was determined by using high-performance liquid chromatography (HPLC). The results showed that surface sterilization of feverfew seeds with 6% NaOCl for 15 min obtained 65.00 ± 2.69% aseptic seeds. Murashige and Skoog (MS) medium supplemented with 0.4 mg/L thidiazuron (TDZ) and 2 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) resulted in 86.00 ± 1.72% callus induction. The highest number of shoots (5.00 ± 0.15) per explant was obtained in the treatment of MS medium supplemented with 5 mg/L zeatin. MS medium fortified with 3 mg/L indole-3-butyric acid (IBA) produced the maximum number of roots per plantlet (8.90 ± 0.35). A total of 90% of the micropropagated plantlets survived when planted in perlite + peat moss (1:1 v/v); the micropropagated plantlets were successfully established in the ex vitro conditions. According to parthenolide analysis, its level was significantly higher in the micropropagated plants than conventionally grown plants. Among different solvents, ethanolic extraction obtained the highest parthenolide content of the feverfew plant. Hence, it can be concluded that micropropagation of feverfew could be applied to produce disease-free planting materials and to improve the parthenolide content of the feverfew plant.

Intestinal Organoids: New Tools to Comprehend the Virulence of Bacterial Foodborne Pathogens

Foodborne diseases cause high morbidity and mortality worldwide. Understanding the relationships between bacteria and epithelial cells throughout the infection process is essential to setting up preventive and therapeutic solutions. The extensive study of their pathophysiology has mostly been performed on transformed cell cultures that do not fully mirror the complex cell populations, the in vivo architectures, and the genetic profiles of native tissues. Following advances in primary cell culture techniques, organoids have been developed. Such technological breakthroughs have opened a new path in the study of microbial infectious diseases, and thus opened onto new strategies to control foodborne hazards. This review sheds new light on cellular messages from the host–foodborne pathogen crosstalk during in vitro organoid infection by the foodborne pathogenic bacteria with the highest health burden. Finally, future perspectives and current challenges are discussed to provide a better understanding of the potential applications of organoids in the investigation of foodborne infectious diseases.

Aklimatisasi dan Respon Pertumbuhan Mutan Leucaena Leucocephala Varietas Tarramba Teradaptasi Asam

Acclimatization is the final stage of plant propagation in tissue culture techniques that can determine the success of the nursery process. This study aimed to observe the growth response during the acclimatization stage of the acid-adapted of Leucaena leucocephala Tarramba variety, which developed from tissue culture techniques. The design used in this study was a completely randomized design (CRD) with the cultivation of 11 mutant lines from tissue culture, namely M1-M11 (an acid-adapted mutant from tissue culture addition of 1 ppm IBA) and 2 controls (lamtoro broodstock without gamma irradiation) namely K0 (lamtoro broodstock resulting from tissue culture addition of 0 ppm IBA), K1 (breeding lamtoro from tissue culture addition of 1 ppm IBA). The variables observed were the level of plant viability, plant height, and number of leaves. The results showed that the acclimatization of the plant Leucaena leucocephala to tissue culture production on the M3 and M9 mutant lines gave the best response to plant morphological growth up to 5 WAP (weeks after planting). Key words: acclimatization, IBA hormone, Leucaena leucocephala, tissue culture

Gradient biomimetic platforms for neurogenesis studies

There is a need for the development of new cellular therapies for the treatment of many diseases, with the central nervous system (CNS) currently an area of specific focus. Due to the complexity and delicacy of its biology, there is currently a limited understanding of neurogenesis and consequently a lack of reliable test platforms, resulting in several CNS based diseases having no cure. The ability to differentiate pluripotent stem cells into specific neuronal sub-types may enable scalable manufacture for clinical therapies, with a focus also on the purity and quality of the cell population. This focus is targeted towards an urgent need for the diseases that currently have no cure, e.g. Parkinson’s disease. Differentiation studies carried out using traditional 2D cell culture techniques are designed using biological signals and morphogens known to be important for neurogenesis in vivo. However, such studies are limited by their simplistic nature, including a general poor efficiency and reproducibility, high reagent costs and an inability to scale-up the process to a manufacture-wide design for clinical use. Biomimetic approaches to recapitulate a more in vivo-like environment are progressing rapidly within this field, with application of bio(chemical) gradients presented both as 2D surfaces and within a 3D volume. This review focusses on the development and application of these advanced extracellular environments particularly for the neural niche. We emphasise the progress that has been made specifically in the area of stem cell derived neuronal differentiation. Increasing developments in biomaterial approaches to manufacture stem cells will enable the improvement of differentiation protocols, enhancing the efficiency and repeatability of the process with a move towards up-scaling. Progress in this area brings these techniques closer to enabling the development of therapies for the clinic.

An improved organ explant culture method reveals stem cell lineage dynamics in the adult Drosophila intestine

In recent years, live-imaging techniques have been developed for the adult midgut of Drosophila melanogaster that allow temporal characterization of key processes involved in stem cell and tissue homeostasis. However, current organ culture techniques are limited to imaging sessions of <16 hours, an interval too short to track dynamic processes such as damage responses and regeneration, which can unfold over several days. Therefore, we developed a new organ explant culture protocol capable of sustaining midguts ex vivo for up to 3 days. This was made possible by the formulation of a culture medium specifically designed for adult Drosophila tissues with an increased Na+/K+ ratio and trehalose concentration, and by placing midguts at an air-liquid interface for enhanced oxygenation. We show that midgut progenitor cells can respond to gut epithelium damage ex vivo, proliferating and differentiating to replace lost cells, but are quiescent in healthy intestines. Using ex vivo gene induction to promote stem cell proliferation, we demonstrate that intestinal stem lineages can be traced through multiple cell divisions using live imaging. Both asymmetric and symmetric divisions can be identified in the reconstructed lineages. We find that daughter cells of asymmetric divisions remain in close proximity of each other, while the progeny of symmetric divisions actively move apart, with implications for cell differentiation and tissue organization. We show that the same culture set-up is useful for imaging adult renal tubules and ovaries for up to 72 hours. By enabling both long-term imaging and real-time ex vivo gene manipulation, our simple culture protocol provides a powerful tool for studies of epithelial biology and cell lineage behavior.

Efficiency of in vitro culture techniques applied to soybean (Glycine max (L.) Merr.) accessions from the VIR collection

Using a wide range of modern biotechnologies and genetic techniques to study plant germplasm accessions held by VIR makes it possible to procure valuable results, required for the development of new high-yielding cultivars adapted to adverse environmental conditions and possessing specified technological properties, particularly to identify and mark new genes and alleles useful for plant breeding. This research trend is in line with Presidential Decree No. 680 “Concerning the development of genetic technologies in the Russian Federation”. Soybean is among the key crops in agricultural production, but the use of next-generation breeding tools to obtain new soybean cultivars with desired properties is still limited. Successful application of novel methods also requires new approaches to studying soybean accessions, specifically their ability to regenerate and produce calluses for subsequent inclusion in biotechnological programs.Ten soybean accessions of various origin, contrasting in ripening schedules, were selected to study the possibility of effective introduction into in vitro culture and further assessment of their ability to produce calluses and regenerate in in vitro culture. The work included evaluating the effects of different seed sterilization techniques (one-step sterilization, using a commercial bleach, and two-step one, combining the impacts of a chlorine-containing preparation and hydrogen peroxide), types of explants (epicotyls, hypocotyls, cotyledon nodes, and cotyledon leaf segments), and phytohormone composition of nutrient medium: (1) MS + 1.13 mg/L BAP + 0.5 mg/L HA, and (2) MS +1 mg/L BAP + 0.1 mg/L IAA).The assessment results showed that the option of two-step seed sterilization was the most effective for soybean at the stage of in vitro culture initiation, while hypocotyls, epicotyls, and cotyledon nodes had the highest callus formation ability in both types of nutrient media.

IN VITRO PROPAGATION OF PLUM ROOTSTOCK DOCERA 6

This paper describes research on the application of tissue culture techniques to the micropropagation of interspecific rootstock ʹDocera 6ʹ. The experimental work was carried out in the period 2017-2018, in the in vitro propagation laboratory of the Fruit Growing Institute Plovdiv. Аxillary buds were employed as initial explantеs in two different seasons (March-May; September-October). The action of the mineral medium was studied in the multiplication stage. The best result was obtained on LS medium included BAP 0.5 mg/l and IAA 0.05 mg/l. Тhe obtained average multiplication rate is 3.08. The concentration of auxin applied to the basal medium influence the quality of the root system Treatment with high concentrations of IBA added to the rooting medium gives the best results (V5). Тhe influence of the season on growth and development of micropropagated ʹDocera 6ʹ rootstock during ex vitro acclimatization is also part of our research. The spring acclimatization gives better results than the autumn.

Occurrence of Escherichia coli in cloacal samples of broiler chicken from Kollam and Kottayam districts

Foodborne pathogens like E. coli are considered as the major causes of foodborne illness in humans worldwide. The present study was undertaken to determine the occurrence of E. coli in cloacal samples of broiler chicken from Kollam and Kottayam districts. The occurrence of E. coli in cloacal samples from broiler chicken was 76.5 per cent from Kollam and 79 per cent from Kottayam through culture techniques. Out of the total 400 cloacal swab samples collected from broiler chicken, 77.8 per cent were positive for E. coli. The samples which were subjected to conventional culture techniques were further analysed for PCR confirmation. The study revealed that, 56.5 and 67 per cent samples were positive for E. coli from Kollam and Kottayam, respectively. An overall occurrence of 61.8 per cent out of 400 samples were confirmed for E. coli by PCR. One Health approach can be used as a suitable tool to combat the foodborne zoonotic diseases, since it is an integrated, multidisciplinary, holistic approach. Proper implementation of biosecurity measures in farms is mandatory to control foodborne zoonotic diseases.