COMPARISON OF MOLECULAR AND GENETIC PROPERTIES OF PINE (Pinus sylvestris L.) SEED PLANTATIONS IN BOSNIA AND HERCEGOVINA

U radu je analizirana je genetička struktura dvije sjemenske plantaže običnog bora (Pinus sylvestris L.) sa 40 klonova (Kozji grm) i 20 klonova (Šamin gaj). Za analizu su uporabljeni izoenzimski biljezi, ukupno 9 enzimskih sustava, na ukupno 12 gen lokusa. Ukupno 55 alela, u obje klonske sjemenskim plantaže. U multiklonskoj plantaži \u27\u27Koziji grm\u27\u27 registrirano 45, a u sjemenskoj plantaži \u27\u27Šamin gaj\u27\u27 35 alela različitih alela. Genetička raznolikost procijenjena je srednjim brojem alela po lokusu (A/L). Dobivena je vrijednost od 3,83 za sjemensku plantažu Kozji grm i 2,91 za plantažu Šamin gaj. Srednji broj genotipova po lokusu (G/L) je iznosio 5,16 za sjemensku plantažu Kozji grm i 3,58 za sjemensku plantažu Šamin gaj. Srednja stvarna heterozigotnost klonova u sjemenskoj plantaži Kozji grm iznosi 0,2833, a u sjemenskoj plantaži Šamin gaj srednja heterozigotnost je 0,3166. Srednja teorijska heterozigotnost za plantažu Kozji grm iznosi 0,3366, a za sjemensku plantažu Šamin gaj 0,3491.The paper analyses the genetic structure of the pine (Pinus sylvestris L.) clones in the multiclone seed plantations of Kozji Grm (KG) and Šamin Gaj (ŠG). The clones in both seed plantations originate from natural populations of common pine in Bosnia and Hercegovina; KG (40 clones): Gornji Janj – 10 clones, Klekovača 7, Kaljina Bioštica 16, Romanija Glasinac 6, Igman 1, while the ŠG plantation contains 20 clones from one population (Igman). The genetic structure was analysed using 9 enzyme systems upon altogether 12 gene loci. Altogether 55 different alleles were observed: in KG plantation 45 alleles were seen, while in ŠG plantation there were 35 alleles. To estimate the genetic diversity, the mean number of alleles per locus (A/L) was calculated: it is 3.83 for KG, and 2.91 for ŠG, while the mean number of genotypes per locus (G/L) was 5.16 for KG and 3.58 for ŠG. The results confirm the assumption that the number of possible classes with genotypes is higher than with alleles. The differences are the most conspicuous if the differences are observed in the occurrence of the individual alleles and the allele frequencies. The mean real hetero-zygote property of the clones in KG plantation is 0.2833, while the same value in ŠG is 0.3166. The mean theoretical hetero/zygote property of KG plantation is 0.3366, and of ŠG plantation it is 0.3491. The present differences show a deviation of the real status from the status of genetic balance according to the Hardy/Weinberg law. The results show that the use of the seed produced in the multiclone seed plantation, which originates from the clones from different provenances, may increase the risk of introducing undesired alleles into the natural population. A more acceptable solution would therefore be the seed plantations from the clones of a single population, and the use of their seed for the regeneration of this population

The Analysis of Genetic Variability of Norway Spruce (Picea abies (L.) Karst.) Subpopulation at the Igman Mountain

Uporabom 12 izoenzimskih sustava, te analizom 20 gen lokusa, uspoređivana je genetička struktura dviju subpopulacija s planine Igman. Prema ekološkim pokazateljima, a misli se na klimu i njeno djelovanje, postoje razlike između subpopulacija, jer jedna pripada mrazištu a druga tipičnoj planinskoj klimi. Rezultati analize izoenzimskih biljega upućuju na postojanje razlika između analiziranih subpopulacija.In this study we analyzed the genetic structure of two autochthonous subpopulations of Norway spruce in the Mountain of Igman by usage of isoenzyme markers. We collected the material for the analysis in two separate plant communities. The subpopulation Igman – A is represented by fir-tree and spruce forest with randomly distributed white pine trees (Abieti-Piceetum illyricum Stef. 1960) while the Igman – B subpopulation is represented through the spruce tree of frosty type in the mountain area (Piceetum montanum s.lat. (Fuk. et Stef., 1958, emend. Horv. et al., 1974)). Between the subpopulations there is a 150 m difference in altitude. We analyzed the following systems of Acotinase (Aco-A), Glutamate dehydrogenase (Gdh-A), Glutamate oxaloacetate transaminase (Got-A, Got-B, Got-C), Izocitrate dehidrogenase (Idh-A, Idh-B), Leucine aminopeptidase (Lap-B), Malate dehydrogenase (Mdh-A, Mdh-B, Mdh-C), Menadione reductase (Mnr-A, Mnr-C), Phosphoglucose isomerase (Pgi-B), Phosphoglucomutase (Pgm-A), Shikimate dehydrogenase (Skdh-A), 6-Phosphogluconate dehydrogenase (6-Pgdh-A, 6-Pgdh-B, 6-Pgdh-C) and Fluorescentesterase (Fest-B). The frequency of the allele and the frequency of genotypes show diversity between subpopulations. The Allele differentiation was most evident at loci Got-C, 6-Pgdh-A. In the sample of the Igman – A subpopulation the frequency of the allele Aco-A2 was 7 % lower, and the frequency of 6-Pgdh-A2 7 % higher than in the sample from Igman – B subpopulation. The genotype subpopulations are most explicitly differentiated at loci Fest-B, Got-C, Lap-B, Mdh-C, Mnr-A, Mnr-C, Pgi-B, 6-Pgdh-A, 6-Pgdh-B, 6-Pgdh-C. If the Igman – A subpopulation is compared with Igman – B subpopulation, we have 8–14% higher frequency of homozygote: Got-C44 (52 % vs. 44 %), Fest22 (90 % vs. 80 %), Mnr-A22 (12 % vs. 4 %), Mnr-C22 (94 % vs. 82 %), 6-Pgdh-A22 (94 % vs. 80 %), 6-Pgdh-C22 (42 % vs. 30 %) and from 10–14 % higher heterozygote frequency for gene loci: Lap-B46 (12 % vs. 0 %), Pgi-B23 (52 % vs. 42 %), 6-Pgdh-B25 (54 % vs. 40 %). In Igman – B subpopulation versus Igman – A subpopulation has 10 % higher homozygote frequency, as follows: Pgi-B33 (46 % vs. 36 %), 6-Pgdh-B22 (50 % vs. 40 %) and between 8–14 % heterozygote frequency Fest-B12 (14 % vs. 2 %), Mnr-A24 (70 % vs. 56 %), Mnr-C23 (16 % vs. 4 %), 6-Pgdh-A23 (12 % vs. 4 %), 6-Pgdh-C25 (60 % vs. 46 %). By statistical calculation we obtained an average number of allele per locus, thus in the subpopulation A the number of allele per locus was 2,71, and the effective was 1,307, and in the subpopulation B it was 2,59, while the effective number was 1,332. The actual heterozygosis in subpopulation A was 24,4 %, and expected was 84,1 %, and in the subpopulation B the actual was 26,2 %, and expected 81,9 %. The number of polymorphous loci in both populations was 17, and the percentage of polymorphous loci was 85,00%. Through the analysis of the allele genetic closeness and genetic distance (d0), we can conclude that the closeness is very high, and differences are relatively small. Thus we determined that the allele closeness has the value of 0,959, and the distance is 0,041 according to Gregorius (1974), which in our case is an extremely high value taking into account the distance between subpopulations of approximately 2 km. Applied statistical parameters for comparison of populations did not show major differences, but the analysis of the direct comparison of the allele presence and their frequency points at the existence of differences, that is, the influence of diverse selection pressures at populations

Geographic variation of Pinus heldreichii Christ from the Western Balkans based on cone and seed morphology

Pinus heldreichii (Bosnian pine) is a Balkan-Apennine endemic and relict pine species that inhabits high mountains in the Mediterranean and sub-Mediterranean regions. Nineteen populations of P. heldreichii from the Western Balkans encompassing 187 individual trees were examined to evaluate morphological variation, a rarely studied aspect of the species. Univariate and multivariate statistics were applied in order to assess the variation of morphological traits of cones and seeds, evaluate the relationships among the sampled populations and verify geographic differentiation in the Dinaric Alps versus Scardo-Pindic mountains. The observations of P. heldreichii covering the populations from the western margins and the centre of the species distribution range indicated a morphological variation among populations and their geographic structure. In general, the southern populations (Scardo-Pindic group) had lower values for the most of morphological traits than the northern ones (Dinaric group). The observed geographic differences between these populations exhibit a north-western to south-eastern gradient, with a few inconsistencies. The southernmost sampled population, Tomorr in Albania, showed remarkable morphological divergence from the other studied populations and appeared to be a distinct morphological group. The pattern of morphological variation in Bosnian pine most likely resulted from multiple effects of long-term isolation and fragmentation in high mountain systems, adaptation to extreme environments and human disturbances

Morphological variability of differentiated by altitude above sea level population of pedunculate oak (Quercus robur L.) in Bosnia and Herzegovina

This study identified altitude differentiation in 44 natural populations of the pedunculate oak in Bosnia and Herzegovina, ranging in altitude from 82 to 860 m. The morphological variation was analyzed on the basis of ten measured and two derived traits, and the populations were divided by altitude into seven groups. The purpose of this study was to determine whether there is statistically significant variation between populations, and to identify the nature of altitude differentiation, as important to preservation and use. Fundamental statistical methods were applied to the analysis, with basic statistical parameters, and a discriminant canonical analysis to determine altitude differentiation in the populations. The results obtained point to statistically significant intra- and inter-popu­lation differences. Altitude-related clinal variation is less pronounced, and is determined largely by the microclimatic factors. These indicators will be taken into account in the formulation of guidelines for good management and restoration of the oak forests and their introduction in foothill and mountain regions, and this experiment is essential to further restore and preserve oak pedunculate using in situ and ex situ methods