Biogeography of Rhaponticoides, an Irano-Turanian element in the Mediterranean flora.

Floristic relationships between the Irano-Turanian and Mediterranean regions have been known from old. However, only a few biogeographical analyses based on molecular data have evaluated the history of steppe plants within the Mediterranean basin. Our study aims to contribute to a better understanding of the migratory and diversification processes by reconstructing the biogeography of Rhaponticoides (Cardueae), distributed in the Mediterranean and Irano-Turanian regions. We generated nuclear and plastid sequences that were analyzed by Bayesian inference. We used the resulting phylogeny for dating the diversification of the genus and examining the dispersal pathways. Two clades were recovered, an Irano-Turanian clade and a Mediterranean clade. The origin of the genus was placed in the Anatolian plateau in the Middle Miocene. The genus experienced several diversifications and expansions correlated to the Messinian salinity crisis and the environmental changes in the Pliocene and the Quaternary. Rhaponticoides migrated following two routes reflecting the two souls of the genus: Irano-Turanian taxa colonized the steppes of Eurasia whilst Mediterranean species migrated via eastern and central Mediterranean and North Africa, leaving a trail of species; both pathways ended in the Iberian Peninsula. Our study also confirms that more work is needed to unravel phylogenetic relationships in Rhaponticoides.

The comparative palynomorphological studies on Turkish Muscari genera and close relatives (Asparagaceae-scilloideae) with their taxonomic implications.

By this study, the palynomorphological traits of 49 taxa (53 plant samples) of the genus Muscari which has been the main research topic and also its relatives, Pseudomuscari, Hyacinthella and Bellevalia have been studied in details of both using light and scanning electron microscope. In the end of palynologic works, it was suggested the descriptive or symbolized main pollen types to make easier the separation or identification of the species of Muscari and its relative groups. According to this symbolization, the pollen types of species may be changeable as D-shaped, elliptical, triangular, and spindle shaped and all types could be seen in different members of the studied taxonomic groups in this paper. One of the most important findings of the study is that the taxa of the genus Muscari, Hyacinthella and Bellevalia do not show a main difference in terms of pollen structures and especially they exhibit harmony. Therefore, it does not possible to mention about the special pollen shape own to one species but it is possible to see common pollen types for closely related taxa or taxon groups. As a result, it could be commented that there is not evolved the close taxonomical groups within Asparagaceae in terms of pollen shape and morphologies and there is considerably a high suitability in point of theirs germ cells or microspores among them. RESEARCH HIGHLIGHTS: A detailed palynological description is provided for Muscari and close relative genus using scanning electron microscopy (SEM) and light microscopy (LM). Pollen size and sulcus length are been determined as the best informative pollen characters to understand Muscari and other studied species.

Phytochemical Analysis, Antioxidant and Anticancer Potential of Sideritis niveotomentosa: Endemic Wild Species of Turkey.

Sideritis niveotomentosa Hub. -Mor. is a local endemic species belonging to the Lamiaceae family. In this study, GC/MS analysis, total antioxidant capacity and anticancer effects of different extracts obtained from S. niveotomentosa were investigated comparatively. Total phenolic contents of extracts were determined by the Folin-Ciocalteu method, total flavonoid contents by aluminum chloride method, and also the free radical scavenging activities of the extracts by DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay. The cytotoxic effect of the extracts was studied via MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay on DLD1, HL60 and ARH77 cell lines. Pro-apoptotic gene expression levels were also tested in the most sensitive cell line ARH77 by Real-Time PCR. The expression levels of 4 pro-apoptotic genes, APAF, BAX, CASP3, and HRK were found to be upregulated in ARH77 cells that were treated extracts. Results showed that methanolic extracts contain more phenolic content than acetone extracts, consistent with DPPH results. As a result, Sideritis niveotomentosa extracts, especially methanolic extracts, are rich in phenolic content and have a strong radical scavenging effect. In addition, the extracts showed selective effects on cell lines. This study is pioneering in terms of future studies, and the findings provide hope for future experimentation.

Speciation and genetic diversity in Centaurea subsect. Phalolepis in Anatolia.

Mountains of Anatolia are one of the main Mediterranean biodiversity hotspots and their richness in endemic species amounts for 30% of the flora. Two main factors may account for this high diversity: the complex orography and its role as refugia during past glaciations. We have investigated seven narrow endemics of Centaurea subsection Phalolepis from Anatolia by means of microsatellites and ecological niche modelling (ENM), in order to analyse genetic polymorphisms and getting insights into their speciation. Despite being narrow endemics, all the studied species show moderate to high SSR genetic diversity. Populations are genetically isolated, but exchange of genes probably occurred at glacial maxima (likely through the Anatolian mountain arches as suggested by the ENM). The lack of correlation between genetic clusters and (morpho) species is interpreted as a result of allopatric diversification on the basis of a shared gene pool. As suggested in a former study in Greece, post-glacial isolation in mountains would be the main driver of diversification in these plants; mountains of Anatolia would have acted as plant refugia, allowing the maintenance of high genetic diversity. Ancient gene flow between taxa that became sympatric during glaciations may also have contributed to the high levels of genetic diversity.

Antioxidant capacity and fatty acid composition of different parts of Adenocarpus complicatus (Fabaceae) from Turkey.

Adenocarpus complicatus is distributed throughout the Anatolian peninsula and is widely used for human and animal nutrition. The purpose of this work was to study the antioxidant properties and fatty acid composition of different parts of this plant (fruits and mixed materials). The species was collected from Golyuzu village of the Seydisehir district near Konya province, Turkey. Fruit and mixed parts obtained from this species were ground and a 15g sample was used to prepare methanolic extracts. Powdered plant samples were extracted with 100mL methanol in a mechanical shaker. The obtained extracts were filtered and concentrated to dryness under reduced pressure and were subsequently stored at -20 degrees C. Antioxidant components, namely total phenolic and flavonoid content, were detected for each extract using spectrophotometric methods. Antioxidant capacity was evaluated by various assays including phosphomolybdenum, DPPH free radical scavenging capacity, metal chelating activity, and ferric and cupric ion reducing power. The fatty acid profiles of plant parts were also determined by using gas chromatography. The total phenolic content of fruit (36.21mgGAE/g) was higher than that of mixed materials (13.79mgGAE/g). The methanolic extract of mixed material had higher amounts of flavonoid than fruit extract. The free radical scavenging activity of extracts was expressed as IC50 value (microg/mL) (amount required to inhibit DPPH radical formation by 50%). The lower IC50 value reflects better free radical scavenging action. The radical scavenging activity of the samples was compared with BHT, it showed the mixed material to be almost two times more potent than the fruit extract. However, BHT is an excellent free radical scavenger with an IC50 of 34.061 microg/mL. The ferric and cupric reducing power potentials of the extracts were expressed as EC50 value (the effective concentration at which the absorbance was 0.5). Fruit extract exhibited strong ferric reducing power with an EC50 of 871.25 microg/mL. The metal chelating activity of the extracts increased with concentration. Chelating effect was 83.60% for fruit extract at 1mg/mL concentration. Oil content of fruit and mixed parts were detected as 6.71 and 6.14%, respectively. A total of 32 fatty acids were found in the oil. Essential fatty acids (linoleic and a-linolenic acid) were identified as the most abundant fatty acids in the oil. These results demonstrated that this plant species can be considered as an alternative to synthetic antioxidants. Likewise, the oil obtained from the plant can be used as a source of essential fatty acids for food and pharmacological applications.

Phylogeny of the Centaurea group (Centaurea, Compositae) - geography is a better predictor than morphology.

The Centaurea group is part of the Circum-Mediterranean Clade (CMC) of genus Centaurea subgenus Centaurea, a mainly Mediterranean plant group with more than 200 described species. The group is traditionally split on morphological basis into three sections: Centaurea, Phalolepis and Willkommia. This division, however, is doubtful, especially in light of molecular approaches. In this study we try to resolve this phylogenetic problem and to consolidate the circumscription and delimitation of the entire group against other closely related groups. We analyzed nuclear (internal transcribed spacer of the ribosomal genes) and chloroplast (rpl32-trnL intergenic spacer) DNA regions for most of the described species of the Centaurea group using phylogenetic and network approaches, and we checked the data for recombination. Phylogeny was used to reconstruct the evolution of the lacerate-membranaceous bract appendages using parsimony. The magnitude of incomplete lineage sorting was tested estimating the effective population sizes. Molecular dating was performed using a Bayesian approach, and the ancestral area reconstruction was conducted using the Dispersal-Extinction-Cladogenesis method. Monophyly of the Centaurea group is confirmed if a few species are removed. Our results do not support the traditional sectional division. There is a high incongruence between the two markers and between genetic data and morphology. However, there is a clear relation between geography and the structure of the molecular data. Diversification in the Centaurea group mainly took place during the Pliocene and Pleistocene. The ancestral area infered for the Circum-Mediterranean Clade of Centaurea is the Eastern Mediterranean, whereas for the Centaurea group it is most likely NW-Africa. The large incongruencies, which hamper phylogenetic reconstruction, are probably the result of introgression, even though the presence of incomplete lineage sorting as an additional factor cannot be ruled out. Convergent evolution of morphological traits may have led to incongruence between morphology-based, traditional systematics and molecular results. Our results also cast major doubts about current species delimitation.

Antioxidant capacity and fatty acid composition of different parts of Adenocarpus complicatus (Fabaceae) from Turkey / Capacidad antioxidante y composición de ácidos grasos de diferentes partes de Adenocarpus complicatus (Fabaceae) de Turquía

Adenocarpus complicatus is distributed throughout the Anatolian peninsula and is widely used for human and animal nutrition. The purpose of this work was to study the antioxidant properties and fatty acid composition of different parts of this plant (fruits and mixed materials). The species was collected from Golyuzu village of the Seydisehir district near Konya province, Turkey. Fruit and mixed parts obtained from this species were ground and a 15g sample was used to prepare methanolic extracts. Powdered plant samples were extracted with 100mL methanol in a mechanical shaker. The obtained extracts were filtered and concentrated to dryness under reduced pressure and were subsequently stored at -20°C. Antioxidant components, namely total phenolic and flavonoid content, were detected for each extract using spectrophotometric methods. Antioxidant capacity was evaluated by various assays including phosphomolybdenum, DPPH free radical scavenging capacity, metal chelating activity, and ferric and cupric ion reducing power. The fatty acid profiles of plant parts were also determined by using gas chromatography. The total phenolic content of fruit (36.21mgGAE/g) was higher than that of mixed materials (13.79mgGAE/g). The methanolic extract of mixed material had higher amounts of flavonoid than fruit extract. The free radical scavenging activity of extracts was expressed as IC50 value (μg/mL) (amount required to inhibit DPPH radical formation by 50%). The lower IC50 value reflects better free radical scavenging action. The radical scavenging activity of the samples was compared with BHT, it showed the mixed material to be almost two times more potent than the fruit extract. However, BHT is an excellent free radical scavenger with an IC50 of 34.061μg/mL. The ferric and cupric reducing power potentials of the extracts were expressed as EC50 value (the effective concentration at which the absorbance was 0.5). Fruit extract exhibited strong ferric reducing power with an EC50 of 871.25μg/mL. The metal chelating activity of the extracts increased with concentration. Chelating effect was 83.60% for fruit extract at 1mg/mL concentration. Oil content of fruit and mixed parts were detected as 6.71 and 6.14%, respectively. A total of 32 fatty acids were found in the oil. Essential fatty acids (linoleic and α-linolenic acid) were identified as the most abundant fatty acids in the oil. These results demonstrated that this plant species can be considered as an alternative to synthetic antioxidants. Likewise, the oil obtained from the plant can be used as a source of essential fatty acids for food and pharmacological applications. Rev. Biol. Trop. 62 (1): 337-346. Epub 2014 March 01.

Adenocarpus complicatus se distribuye por toda la península de Anatolia y es ampliamente utilizado para la nutrición humana y animal. El propósito de este trabajo fue estudiar las propiedades antioxidantes y la composición de ácidos grasos de diferentes partes de la planta (frutos y partes mezcladas). Las especies fueron recolectadas en Golyuzu, Seydisehir, cerca de la provincia Konya en Turquía. Para preparar los extractos metanólicos se tomó una muestra de 15g de frutas y partes mezcladas de esta especie. Muestras de plantas en polvo se extrajeron con 100ml de metanol en un agitador mecánico. Los extractos obtenidos se filtraron y se concentraron a sequedad bajo presión reducida y posteriormente se almacenaron a -20°C. Para cada extracto, mediante métodos espectrofotométricos se detectaron los componentes antioxidantes, llamados contenido total de fenoles y flavonoides. La capacidad antioxidante se evaluó mediante diversos ensayos: fosfomolibdeno, capacidad de captación de radicales libres DPPH, actividad quelante de metales y poder reductor de iones férricos y cúpricos. También se determinaron los perfiles de ácidos grasos de partes de la planta mediante el uso de cromatografía de gases. El contenido fenólico total de la fruta (36.21mgGAE/g) fue mayor que la de los materiales mezclados (13.79mgGAE/g). El extracto metanólico de material mezclado tenía una mayor cantidad de flavonoides que el extracto de la fruta. La actividad captadora de radicales libres de los extractos se expresó como valor de IC50 (mg/ml) (cantidad necesaria para inhibir la formación de radicales DPPH en un 50%). El valor bajo de IC50 refleja mejor acción eliminadora de radicales libres. La actividad captadora de radicales de las muestras se comparó con BHT, se mostró que el material mezclado es casi dos veces más potente que el extracto de la fruta. Sin embargo, BHT es un excelente eliminador de radicales libres con una IC50 de 34.061μg/mL. El potencial de reducción férrico y cúprico de los extractos se expresó como valor de CE50 (la concentración efectiva a la que la absorbancia fue de 0.5). El extracto de la fruta exhibe fuerte poder reductor férrico con una EC50 de 871.25μg/mL. La actividad quelante de metales de los extractos aumentó con la concentración. El efecto quelante de extracto de fruta fue de 83.60% en una concentración de 1mg/ml. El contenido de aceite del fruto y partes mixtas fue 6.71 y 6.14%, respectivamente. Un total de 32 ácidos grasos fueron encontrados en el aceite. Los ácidos grasos esenciales (ácido linoleico y α-linolénico) fueron identificados como los ácidos grasos más abundantes en el aceite. Estos resultados demostraron que esta especie vegetal se puede considerar como una alternativa a los antioxidantes sintéticos. Del mismo modo, el aceite obtenido de la planta se puede utilizar como una fuente de ácidos grasos esenciales para alimentos y aplicaciones farmacológicas.

In vitro investigation of cytotoxic and apoptotic effects of Cynara L. species in colorectal cancer cells.

Apoptotic and cytotoxic activity of plant extracts obtaining from naturally growing Cynara syriaca in Turkey and cultivated C cardunculus against DLD1 colorectal cancer cells was determined. Extracts from wild and cultivated Cynara species were obtained from their vegetative parts and receptacles using hexane and applied with five different dose (0.1-1 mg/ml) as well as apigenin for MTT tests for three time periods (24, 48 and 72 hours). After cells were treated with IC50 doses for each extract total DNA and RNA were isolated for determination of the cause of cell death. From isolated RNAs, cDNA were synthesized and amplification of p21, BCL-2 and BAX gene regions was carried out. Consequently, we found that pro-apoptotic (BAX) gene expression and a cell cycle inhibitor (p21) were induced in the presence of our artichoke extracts. In contrast, anti-apoptotic BCL-2 gene expression was reduced compared to the control group. In addition DNA fragmentation results demonstrated DLD1 cell death via apoptosis.