Supplementary data for article: Dodoš, T.; Rajčević, N.; Tešević, V.; Marin, P. D. Chemodiversity of Epicuticular N-Alkanes and Morphological Traits of Natural Populations of Satureja Subspicata Bartl. Ex Vis. along Dinaric Alps – Ecological and Taxonomic Aspects. Chemistry and Biodiversity 2017, 14 (2). https://doi.org/10.1002/cbdv.201600201

Supplementary material for: [https://doi.org/10.1002/cbdv.201600201]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2416

Cross-species amplification of nuclear est-microsatellites developed for other pinus species in pinus nigra

Due to the current lack of nuclear microsatellites (simple sequence repeats SSRs) specifically developed for Pinus nigra, an important European coniferous species, we cross-species amplified 12 EST-SSRs (expressed sequence tagged SSRs) developed for other Pinus species in P. nigra in order to delineate loci which can be used for assessing levels of genetic diversity and genetic structuring in this species. We amplified these loci in individuals from seven populations from the central Balkans representing four recognized infraspecific taxa of P. nigra (ssp. nigra, var. gocensis, ssp. pallasiana, and var. banatica). Contrary to expectations on high transferability of EST-SSRs into related species, only three out of 12 tested loci were successfully amplified in P. nigra, but they displayed lack/low levels of polymorphism or generated multilocus amplification products. Thus, our estimates on levels of genetic diversity (HE = 0.183) and genetic differentiation (F-ST = 0.007) were based on variability of a single locus harboring four alleles only and they should be taken with cautions. Our study highlights the need for the development of high-resolution molecular markers, such as co-dominant genic or genomic SSRs or predominantly biallelic SNPs, or utilization of anonymous dominant markers, such as AFLPs, for genotyping in P. nigra

Genetic patterns in Pinus nigra from the central Balkans inferred from plastid and mitochondrial data

Pinus nigra J.F. Arnold, European black pine, is a typical component of Mediterranean and sub-Mediterranean coniferous forests with highly fragmentary distribution. Western Mediterranean populations of this species have been studied genetically to date, while eastern populations from the central Balkans, which are larger and more abundant, are still genetically understudied. We analyzed seven populations of P. nigra representing all infraspecific taxa recognized within the central Balkans (subspecies nigra with varieties nigra and gocensis Dordevic; and subspecies pallasiana (Lamb.) Holmboe with varieties pallasiana and banatica (Endl.) Georgescu et Ionescu), with three chloroplast microsatellites (cpDNA SSRs) and one mitochondrial (mtDNA) locus. Although our molecular data failed to support circumscription of studied infraspecific taxa, we found that genetic patterns at both genomes are in accordance with those found previously in westward populations of this species, that is - exceptionally high levels of genetic diversity (H-T = 0.949) and low genetic differentiation (G(ST) = 0.024) at the cpDNA level, and moderate levels of genetic diversity (H-T = 0.357) and genetic differentiation (G(ST) = 0.358) at the mtDNA level. Based on genealogical relations of mtDNA types currently present in Balkans' and Iberian/African populations, we inferred that the ancestral gene pool of P. nigra already harbored polymorphism at position 328 prior to the divergence to two lineages currently present in westward and eastward parts of the species range distribution. Subsequent occurrence of three mutations, which distinguish these two lineages, suggests their long-term isolation

Genetic patterns in Pinus nigra from the central Balkans inferred from plastid and mitochondrial data

Pinus nigra J.F. Arnold, European black pine, is a typical component of Mediterranean and sub-Mediterranean coniferous forests with highly fragmentary distribution. Western Mediterranean populations of this species have been studied genetically to date, while eastern populations from the central Balkans, which are larger and more abundant, are still genetically understudied. We analyzed seven populations of P. nigra representing all infraspecific taxa recognized within the central Balkans (subspecies nigra with varieties nigra and gocensis Dordevic; and subspecies pallasiana (Lamb.) Holmboe with varieties pallasiana and banatica (Endl.) Georgescu et Ionescu), with three chloroplast microsatellites (cpDNA SSRs) and one mitochondrial (mtDNA) locus. Although our molecular data failed to support circumscription of studied infraspecific taxa, we found that genetic patterns at both genomes are in accordance with those found previously in westward populations of this species, that is - exceptionally high levels of genetic diversity (H-T = 0.949) and low genetic differentiation (G(ST) = 0.024) at the cpDNA level, and moderate levels of genetic diversity (H-T = 0.357) and genetic differentiation (G(ST) = 0.358) at the mtDNA level. Based on genealogical relations of mtDNA types currently present in Balkans' and Iberian/African populations, we inferred that the ancestral gene pool of P. nigra already harbored polymorphism at position 328 prior to the divergence to two lineages currently present in westward and eastward parts of the species range distribution. Subsequent occurrence of three mutations, which distinguish these two lineages, suggests their long-term isolation.Ministry of Education, Science, and Technology of the Republic of Serbia {[}173029, 173030

n-alkanes in the needle waxes of Pinus heldreichii var. pančići

This is the first report of n-alkanes in needle epicuticular waxes of the variety Bosnian pine, Pinus heldreichii var. pančići. n-Hexane extracts of needle samples, originating from seven isolated localities in Serbia, were analysed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The results evidenced n-alkanes ranging from C18 to C33 in epicuticular waxes. The most abundant alkanes were C27, C23, C25 and C29 (12.53 %, 12.46 %, 12.00 % and 10.38 % on average, respectively). The carbon preference index (CPItotal) of Pinus heldreichii var. pančići ranged from 1.1 to 2.1 (1.6 on average), while the average chain length (ACLtotal) ranged from 25.0 to 25.8 (25.3 on average). A high level of individual quantitative variation in all of these hydrocarbon parameters was also found. The obtained results were compared with the bibliographic references for Pinus heldreichii var. leucodermis and other species of the Pinus genus.Ovo je prvo saopštenje o n-alkanima u voskovima iglica varijeteta munike, Pinus heldreichii var. pančići. n-Heksanski ekstrakti uzoraka iglica koji potiču sa sedam izolovanih lokaliteta u Srbiji analizirani su gasnom hromatografijom (GC) i gasnom hromatografijom-masenom spektrometrijom (GC-MS). Rezultati su pokazali da se n-alkani nalaze u opsegu od C18 do C33. Među njima su najobilniji C27, C23, C25 i C29 (12,53, 12,46, 12,00 i 10,38 % u proseku, redom). Ugljenični preferencijalni indeks (CPItotal) Pinus heldreichii var. pančići je bio u opsegu od 1,1 do 2,1 (prosečno 1,6), a dužina niza ugljenikovih atoma (ACLtotal) u opsegu od 25,0 do 25,8 (prosečno 25,3). Takođe je utvrđen visok nivo individualne kvantitativne varijabilnosti u svim analiziranim parametrima ovih ugljovodonika. Dobijeni rezultati su upoređeni sa literaturnim podacima koji se odnose na Pinus heldreichii var. leucodermis i druge vrste roda Pinus.Projekat ministarstv

Supplementary data for the article: Nikolić, B.; Matović, M.; Todosijević, M.; Stanković, J.; Cvetković, M.; Marin, P. D.; Tešević, V. Volatiles of Tanacetum Macrophyllum Obtained by Different Extraction Methods. Natural Product Communications 2018, 13 (7), 891–893. https://doi.org/10.1177/1934578x1801300727

Supplementary material: [https://doi.org/10.1177/1934578X1801300727]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2169

Prunus spinosa L. leaf extracts: polyphenol profile and bioactivities

Prunus spinosa leaf extracts in solvents of different polarity (water, ethanol and acetone), their phenol, flavonoid and anthocyanin contents and biological properties were the object of this study. The richest in phenols as well as in flavonoids was acetone extract with 181.19 mg GAE and 80.10 mg QE per gram of dry extract, respectively. Moreover, the quantity of anthocyanins obtained by HPLC analysis was also the highest in acetone sample. Examined samples possessed antioxidant properties evaluated through four in vitro assays (DPPH, ABTS, FRAP and TRC). The acetone extract was proved to be the best antioxidant among tested samples, which could be ascribed to polyphenols, especially anthocyanins. The aqueous and the ethanol extract exhibited antibacterial effects, being particularly active against B. cereus and E. cloacae. T. viride, P. funiculosum, P. ochrochloron, P. verrucosum var. cyclopium were the most susceptible among fungal microorganisms examined. Both, the aqueous and the ethanol extract expressed inhibitory activity towards enzymes linked to diabetes mellitus type II. Additionally, the ethanol extract showed significantly higher potential in inhibiting α-glucosidase than the drug used as the positive control. Furthermore, the aqueous sample revealed antitumor effects on following malignant cell lines: HeLa, K562 and MDA-MB-453. The results presented herein suggest that P. spinosa leaves should be considered as a natural source of bioactive compounds with potential application in phytopharmacy and food industry

Population diversity of n-alkanes in the needle cuticular wax of relicts pinus heldreichii and P. peuce from the scardo-pindic mountains

The variability of n-alkanes in the needle cuticular wax of Pinus heldreichii and P. peuce in two natural populations from the Scardo-Pindic mountains was investigated for the first time. Gas chromatography (GC) and gas chromatography/mass spectrometry (GC-MS) analyses of two-year-old needles were performed us-ing an Agilent 7890A GC equipped with an inert 5975C XL EI/CI mass spectrometer detector (MSD) and a flame ionization detector (FID) connected by a capillary flow technology 2-way splitter with make-up. An HP-5MS capillary column was used. n-Alkanes ranged from C19 to C33. In P. heldreichii the most abundant were C23, C25 and C27 (16.3, 15.6 and 12.8 % on average, respectively), while in P. peuce they were C29, C25, and C27 (16.5, 15.3 and 13.5 % on average, resp.). Mid-chain and long-chain n-alkanes prevailed in both species. Principle component analysis (PCA) and Cluster analyses of five and six n-alkanes, respectively, showed divergence of the Scardo-Pindic populations from the Dinaric ones

Comparative anatomical investigation of five Artemisia L. (Anthemideae, Asteraceae) species in view of taxonomy

Comparative anatomical analysis of vegetative organs has been conducted on Artemisia campestris L., A. absinthium L., A. arborescens L., A. judaica L. and A. herba-alba Asso, using light microscopy, in order to examine the most important anatomical features and to find new valid taxonomic characters. Results have shown that general root, stem and leaf anatomical features and nonglandular and glandular trichomes are shared by all species. However, some characters (parenchyma sheath, which surrounded vascular bundle and extended to both epidermises, subepidermal collenchyma and the absence of secretory canals in the leaves) link together A. absinthium and A. arborescens from the same section. Some characters, as periderm and lignified pith parenchyma cells (A. campestris and A. arborescens), nonendodermal secretory canals in root cortex (A. absinthium and A. judaica) and secretory canals in the leaf phloem (A. judaica and A. herba-alba), connect species belonging to different sections. Moreover, some characters could be considered as species-specific, nonendodermal secretory canals in the root secondary phloem, triangular leaf shape on the cross section and secretory canals in the leaf parenchyma for A. campestris, secretory canals in the stem pith for A. absinthium, crystals in the pith parenchyma cells for A. arborescens and the absence of root secretory canals for A. herba-alba. Given results revealed qualitative characters, on the basis of which the studied species are anatomically distinguishable between each other, provide valuable features for better species identification and contribute to the anatomy of the genus Artemisia

Supplementary data for article: Dodoš, T.; Rajčević, N.; Tešević, V.; Marin, P. D. Chemodiversity of Epicuticular N-Alkanes and Morphological Traits of Natural Populations of Satureja Subspicata Bartl. Ex Vis. along Dinaric Alps – Ecological and Taxonomic Aspects. Chemistry and Biodiversity 2017, 14 (2). https://doi.org/10.1002/cbdv.201600201

Supplementary material for: [https://doi.org/10.1002/cbdv.201600201]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2416