High Level of Phenotypic Differentiation of Common Yew (Taxus baccata L.) Populations in the North-Western Part of the Balkan Peninsula

Common or English yew (Taxus baccata L., Taxaceae) is a conifer species, native to Europe, northern Africa, Asia Minor and Caucasus. It is a dioecious, wind-pollinated and animal-dispersed tree, known for its high-quality wood and medicinal properties, albeit poisonous. The species is rare and has been legally protected at the European and national levels. In addition, its low population density and disjunct character of distribution have reinforced the need for its protection as regeneration is mostly lacking. The aim of this study was to phenotypically characterise the north-western Balkan yew populations. Phenotypic diversity was examined for seven populations, using morphometric analysis of nine phenotypic traits of needles. Descriptive and multivariate statistical analyses were conducted to evaluate the inter- and intrapopulation variability. In addition, to test correlations between geographic, climatic and phenotypic data, Mantel test was used. We identified a geographic structure across studied populations that exhibited high levels of variability on intra- and interpopulation levels. Two groups of populations have been defined and are consistent with previously described genetic divergent lineages from separate refugia. In addition, a significant correlation between phenotypic and geographic data were revealed, i.e., isolation by distance (IBD). However, the Mantel test revealed no significant correlation between morphometric and environmental data. In conclusion, our data reveal that the historical events and persistent IBD acted in combination to produce the morphological patterns observed in common yew populations in the north-western part of the Balkan Peninsula. Finally, we suggested conservation measures to be implemented on a stand level, with habitat preservation as the main goal. In addition, ex situ conservation should be considered, both in the form of collections and urban planting, as both provide additional gene pool reserves.

Evaluation of Antimicrobial Activity of Several Medicinal Plants Against Clinically Isolated Staphylococcus Aureus from Humans and Sheep

Background: Continuous and indiscriminate use of chemical drugs causes an important phenomenon of resistance to microorganisms. Accordingly, the effect of medications is minimized or offset, increasing drug use and the need to study mixtures with more latest and powerful formulations. On the other hand, it has been reported that many plants essential oils have a significant inhibitory effect on pathogenic microorganisms. Objectives: This study aimed to evaluate the antimicrobial activity of some curative herbs against some clinical bacteria of humans and sheep. Methods: The leaves of chicory (Cichorium intybus L.), Hypericum perforatum L., Lavandula angustifolia, Thymus vulgaris L., and Taxus baccata L. were collected and determined in the botanical laboratory of the University of Zabol. Forty grams of dried leaves was used in 400 mL of ethanol (96%) to prepare the ethanolic extract. DPPH was used to determine the activity of reactive oxygen species (ROS) trapping. The antimicrobial effects were studied by the disk diffusion (6 mm) method in Müller-Hinton agar medium according to the method by Bauer et al. Results: The minimum inhibitory concentrations (MICs) of chicory, thyme, H. perforatum, French lavender, and yarrow extracts in human clinical Staphylococcus aureus were 6.25, 12.5, 3.1, 25, and 6.25 ppm, respectively, but, in sheep, clinical S. aureus, were 12.5, 6.25, 3.1, 12.5, and 25 ppm, respectively. The minimum bactericidal concentrations (MBCs) of chicory, thyme, H. perforatum, French lavender, and yarrow extracts in human clinical S. aureus were 12.5, 25, 6.25, 50, and 12.5 ppm, respectively, but, in sheep clinical S. aureus, were 25, 12.5, 6.25, 25, and 50 ppm, respectively. The most effective extract in inhibiting the growth of S. aureus was the H. perforatum L. extract with an 8.9-mm diameter growth inhibition zone. Conclusions: Regarding the side effects of artificial medications and antimicrobials, as well as the significant influence of healing herb extracts used in this study, it was found that H. perforatum was the most effective plant against S. aureus. It should be noted that plant extracts were more effective in human clinical S. aureus than in sheep clinical S. aureus.

Search for alternative plant raw materials for food industry and environmentally safe animal breeding

The relevance of the search for alternative plant raw materials to enrich the diets of population throughout the world, including in the Russian Federation, is explained by global mass year-round insufficiency of biologically active nutrients in food. This work is aimed at identifying the most promising food plant species from the list, proposed on basis of long-term observations by staff of Tsytsin Main Moscow Botanical Garden of Russian Academy of Sciences (MBG RAS), by bachelors, studying in direction 06.03.01 (Biology) at Moscow State University of Food Production. The research materials (2016-2018) are fruits of hawthorns, rosehips, quince, eleutherococcuses, medlar, mahonia, spindle tree, yews, sumacs, oaks, red-fruited honeysuckles and needles of coniferous plants. Research results indicate the value of studied raw materials as a whole and its advantages in compared species within the genus of plants. Some recommendations for practical usage were made. The following things have been proposed: separation of fruit pulp and seeds before drying of fruits of Crataegus submollis and more preferable C. punctata; wide plantation cultivation of rosehips Rosa amblyotis and R. oxyodon; non-waste separate application of pericarp and seeds of Cydonia oblonga, Mespilus germanica and Mahonia aquifolium; medicinal use of fruits of Eleutherococcus sessiliflorus , and more preferable E. senticosus ; further study of cones of Taxus baccata and T. media; as a spice there is usage of fruits of Rhus typhina ; obtaining unique oils from fruit pulp of Lonicera glehnii, L . ferdinandii, L. maackii and arils of Euonymus europaeus; usage of acorns of Quercus robur, Q. rubra and needles of Picea abies, Pinus sylvestris in feed production. Thus, possibilities of intensifying plant resource usage in the food industry and agriculture were discovered, primarily due to fruits of native introduced species. It is necessary to continue search and further study of the most promising species - sources of alternative plant raw materials.

El censo como herramienta para conocer y proteger La Tejeda del Mosquito

Unos kilómetros al norte del Hayedo de Montejo, en la zona conocida como El Mosquito, la Asociación de Amigos del Tejo y las Tejedas (AATT) ha realizado un censo de Taxus baccata L. en el que se han contabilizado 8.304 ejemplares. Alrededor de la mitad (54%) tiene entre 5 cm y 1,5 m de altura mientras el resto presenta mayor talla, incluso hasta perímetros de tronco superiores a 3 m. La mayor parte se localizan en zonas muy concretas, ocupando el grueso de la población unas 55 ha, y con una distribución estructural y espacial que demuestra la plasticidad de la especie. El tamaño y la densidad de esta población supera la de cualquier otra de la Comunidad de Madrid y del Sistema Central conocidas hasta el momento, y se ha determinado que el área estudiada tiene todas las características exigidas para ser considerada Hábitat 9580*: Bosques mediterráneos de Taxus baccata L., con las peculiaridades en la composición de las especies acompañantes inherentes a los bosques mediterráneos, acentuadas por su influencia atlántica.

Comparative Genomics Analysis of Repetitive Elements in Ten Gymnosperm Species: “Dark Repeatome” and Its Abundance in Conifer and Gnetum Species

Repetitive elements (RE) and transposons (TE) can comprise up to 80% of some plant genomes and may be essential for regulating their evolution and adaptation. The “repeatome” information is often unavailable in assembled genomes because genomic areas of repeats are challenging to assemble and are often missing from final assembly. However, raw genomic sequencing data contain rich information about RE/TEs. Here, raw genomic NGS reads of 10 gymnosperm species were studied for the content and abundance patterns of their “repeatome”. We utilized a combination of alignment on databases of repetitive elements and de novo assembly of highly repetitive sequences from genomic sequencing reads to characterize and calculate the abundance of known and putative repetitive elements in the genomes of 10 conifer plants: Pinus taeda, Pinus sylvestris, Pinus sibirica, Picea glauca, Picea abies, Abies sibirica, Larix sibirica, Juniperus communis, Taxus baccata, and Gnetum gnemon. We found that genome abundances of known and newly discovered putative repeats are specific to phylogenetically close groups of species and match biological taxa. The grouping of species based on abundances of known repeats closely matches the grouping based on abundances of newly discovered putative repeats (kChains) and matches the known taxonomic relations.

Bottleneck Removal of Paclitaxel Biosynthetic Pathway By Overexpression of DBTNBT Gene Under Methyl- β- Cyclodextrin and Coronatine Elicitation in Taxus Baccata L.

Paclitaxel is a highly functionalized diterpenoid that is broadly used for the treatment of several cancer types. This valuable specialized metabolite naturally exists in the inner bark of Taxus species in low amounts. The limited-scale production of paclitaxel in Taxus cell cultures has necessitated the use of several elicitors. Recently, methyl-β-cyclodextrin (CD) and coronatine (COR) have been considered to be highly effective elicitors in producing plant specialized metabolites. Given the limited production of paclitaxel due to the rate limiting enzymes' function, bottleneck removal is conducive to the production of more significant amounts of paclitaxel. In the present study, the full length of DBTNBT coding sequence (CDS), as one of the paclitaxel pathway bottlenecks, was integrated downstream of the CaMV 35S promoter (pCAMBIA1304-DBTNBT) and transiently expressed in Taxus baccata leaves via Agrobacterium tumefaciens and vacuum infiltration method. Paclitaxel production and the expression level of several involved genes were evaluated through different treatments. The transient overexpression of the DBTNBT gene, associated with dual elicitation, resulted in 7.4-fold more paclitaxel production compared with the no-inoculation/no-elicitation control. These ratios were 2.1 and 1.8 in the CD+COR and pCAM treatments, respectively. Among T13αH, T14βH, DBAT, BAPT, DBTNBT, and ABC genes, the most increased expression level belonged to the DBTNBT gene, followed by ABC and BAPT genes. It seems as though in the near future, bottleneck removal could be used on a large scale in Taxus metabolic engineering, resulting in the relative removal of some other bottlenecks and an increase in the final paclitaxel production.

Screening for Selective Anticancer Activity of 65 Extracts of Plants Collected in Western Andalusia, Spain

Finding cytotoxic drugs with a high selectivity towards cancer cells is crucial to improve the low survival rates of patients diagnosed with metastatic cancers. Since plants are an important source of anticancer drugs, we have screened 65 extracts from 45 plants collected in several areas of Western Andalusia (Spain) for cytotoxic activity on lung cancer cells versus lung normal cells. An extract from the leaves of Tetraclinis articulata (Vahl) Mast. (Cupressaceae) showed a marked cytotoxicity (IC50 = 0.37 ± 0.03 μg/mL) and selectivity (selectivity index = 378.3) against the lung cancer cells; cisplatin, 5-fluorouracil, and an extract from the leaves of Taxus baccata L. (Taxaceae) were less cytotoxic and selective. Extracts from Cascabela thevetia (L.) Lippold (Apocynaceae), Frangula alnus Mill. (Rhamnaceae), Iberis ciliata subsp. contracta (Pers.) Moreno (Brassicaceae), Juniperus macrocarpa Sm (Cupressaceae), and Pancratium maritimum L. (Amaryllidaceae) also showed selective cytotoxicity (selectivity index > 10). Active extracts were also tested against a panel of cancer cell lines from a variety tissues. The plants identified in this work are potential sources of natural compounds with selective toxicity towards cancer cells.