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

Significance of diarylheptanoids for chemotaxonomical distinguishing between Alnus glutinosa and Alnus incana

Diarylheptanoids are a group of secondary metabolites widely distributed in the Betulaceae family and characteristic for Alnus species. In this study, the chemotaxonomic power of diarylheptanoids, such as hirsutanonol-5-O-β-d-glucopyranoside, rubranoside A, oregonin, platyphylloside, alnuside A and hirsutanonol, has been investigated in combination with principal component analysis (PCA) for differentiation of Alnus species. Concentrations of six diarylheptanoids in the bark extracts of two natural populations of Alnus glutinosa (black alder) and Alnus incana (gray alder) were determined by ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). PCA clearly shows the separation of three groups. Populations I (A. glutinosa) and II (A. incana) both consisted of individuals of the corresponding species. Four individuals from both sampled populations formed a separate group (population III), which possibly represents a hybrid group. Accordingly, diarylheptanoids may serve in combination with PCA as chemotaxonomic markers at the species level, which may also reveal hybrid species

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

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

n-alkani u voskovima iglica Pinus heldreichii var. pančići

This is the first report of n-alkanes in needle epicuticular waxes of the variety Bosnian pine, Pinus heldreichii var. pancici. 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 C(18) to C(33) in epicuticular waxes. The most abundant alkanes were C(27), C(23), C(25) and C(29) (12.53 %, 12.46 %, 12.00 % and 10.38 % on average, respectively). The carbon preference index (CPI(total)) of Pinus heldreichii var. pancici ranged from 1.1 to 2.1 (1.6 on average), while the average chain length (ACL(total)) ranged from 25.0 to 25.8 (25.3 on average). A high level of individual quantitative variation An 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

Sex expression in Serbian dendroflora: A case study of Fraxinus ornus var. angustifolia

Out of a total number of tree species in Serbian flora, 28% are hermaphrodites, 43% are monoecious and 29% are dioecious. The dendroflora appears to have a larger proportion of unisexual than hermaphrodite species. The monoecious system is the dominant unisexual system within the tree species in Serbia. The possible sex differentiation by RAPD markers and Correspondence Analysis was examined in a population of Fraxinus ornus var. angustifolia. On the basis of 6 selected RAPD bands which contribute to the differentiation in frequency between male and hermaphrodite individuals, Correspondence Analysis visualized the following tendency: 20 trees of each variety were separated into two groups that mainly correspond to sexual types. Our results showed that RAPD markers assisted by Correspondence Analysis could be used in identifying male and hermaphrodite individuals in F. ornus varieties.Projekat ministarstva br. P17301

Diversity of Fraxinus ornus from Serbia and Montenegro as revealed by RAPDs

PCR-RAPD markers revealed individual variation in F. ornus. A total of 122 fragments were amplified using 7 primers and of these 97 fragments were polymorphic. The percentage of polymorphic loci was between 53.3% and 74.6% with an average of 63.1%. The mean gene diversity for all populations was 0.30 and the mean Shannon's index was 0.44. Of the total genetic variation 87% was intra-population whilst 13% was inter-population. The Mantel test revealed significant correlation between genetic and geographical distance matrice. Results herein represent the first use of molecular genetic (DNA) markers to characterize genetic variation in F. ornus populations. The partition of total genetic variance indicates a relatively restricted population differentiation as expected in outcrossing species. Present and future information on genetic structure and variability in F. ornus needs to be incorporated into strategies for the preservation of genetic resources of tree species.U radu su korišćeni PCR-RAPD markeri radi procene individualnih varijacija kod vrste F. ornus. Ukupno 122 fragmenta je amplificirano korišćenjem 7 prajmera, i među njima je bilo 97 polimorfnih fragmenata. Procenat polimorfnih lokusa se kretao između 53.3% i 74.6% sa prosečnom vrednošću od 63.1%. Srednji diverzitet gena za sve ispitivane populacije je iznosio 0.30, dok je srednji Shannon's index imao vrednost 0.44. Od totalne genetičke varijabilnosti 87% pripada intra-populacionoj varijabilnost, a 13% inter-populacionoj. Mantel test je pokazao značajne korelacije između matrica genetičke i geografske distance. Rezultati ovog rada predstavljaju prvu upotrebu molekularno genetičkih (DNA) markera u cilju određivanja genetičke varijabilnosti populacija F. ornus. Odnosi unutar ukupne genetičke varijabilnosti ukazuju na relativno ograničenu populacionu diferencijacije u odnosu na vrednosti koje su očekivane kod stranooplodne vrste. Na osnovu ovih kao i budućih informacija koje se odnose na genetičku strukturu i varijabilnost vrste F. ornus potrebno je kreirati strategije za očuvanje genetičkih resursa drvenastih vrsta.Projekat ministarstva br. 17301

Phytochemical study of the genus Amphoricarpos

Phytochemistry deals with the study of secondary metabolites produced by plants that synthesize these compounds for many reasons, including their own protection against attack of herbivores and plant diseases. Secondary metabolites are believed to represent plant adaptation to various environmental factors and that they enabled the survival of the species. Secondary metabolites of plants can have curative or toxic effects in humans and animals. Herbal medicine has a long tradition in folk medicine and until the early 20th century, when synthetic organic chemistry began to develop, plants were the main source of medicines. The two basic goals of our phytochemical research are: Isolation and identification of new (biologically active) compounds - potential drugs, and chemotaxonomy (chemosystematics). In the following text through one selected example, the genus Amphoricarpos Vis., our phytochemical research is shown on both aspects

Sex expression in Serbian dendroflora: A case study of Fraxinus ornus var. angustifolia

Out of a total number of tree species in Serbian flora, 28% are hermaphrodites, 43% are monoecious and 29% are dioecious. The dendroflora appears to have a larger proportion of unisexual than hermaphrodite species. The monoecious system is the dominant unisexual system within the tree species in Serbia. The possible sex differentiation by RAPD markers and Correspondence Analysis was examined in a population of Fraxinus ornus var. angustifolia. On the basis of 6 selected RAPD bands which contribute to the differentiation in frequency between male and hermaphrodite individuals, Correspondence Analysis visualized the following tendency: 20 trees of each variety were separated into two groups that mainly correspond to sexual types. Our results showed that RAPD markers assisted by Correspondence Analysis could be used in identifying male and hermaphrodite individuals in F. ornus varieties.Projekat ministarstva br. P17301