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Ugotavljanje vpliva gliv na razgradnjo velikih lesnih ostankov (VLO) v pragozdnem rezervatu smo razčlenili na dva dela. Prvi se je ukvarjal z ugotavljanjem zmanjševanja gostot VLO s stopnjo razgradnje, drugi pa z opisom pojavnosti gliv v vzorcih VLO z metodo gelske elektroforeze v gradientu denaturantov (DGGE). Z namenom ugotavljanja zanesljivosti drugačnih, zdravju varnejših metod od živosrebrne metode, tudi na vodnem volumometru. Pri tem je bila pilotno uporabljena tehtnica Sartorius MSA 323S z dodatkom za merjenje gostot YDK01. Merili smo osnovne, nasičene in absolutne gostote. Po pričakovanjih sta nasičeni in osnovni gostoti pokazali trend padanja gostot. Tehtnica Sartorius se je izkazala za zanesljivo in hitro metodo merjenja gostot v vodi, ki pa jo bo potrebno še optimizirati. Molekularne metode ugotavljanja prisotnosti gliv v VLO so v Sloveniji v razvoju. Na podlagi ekstrakcije glivne DNA iz vzorcev VLO smo s pomočjo analitske metode DGGE uspeli pridobiti profile združb gliv za posamezni vzorec. Rezultati so pokazali, da so nekatere glive značilne za začetne stopnje razkroja, druge pa za končne stopnje. Pestrost profilov se s stopnjo razkroja veča, odstopa le stopnja razkroja 1, ki ima nepričakovano veliko stopnjo pestrosti. Razvrstitev vzorcev glede na podobnost DGGE profilov ni bila skladna z razvrstitvijo v stopnje razgradnje, variabilnosti med vzorci znotraj stopenj razgradnje je bila visoka. Povezave med genetskimi in geografskimi razdaljami niso bile ugotovljene. Researching the role of fungi in the decay of coarse woody debris (CWD) of European beech was divided in two parts. Determining the loss of CWD density during decay was done in part one, while the second part investigated the distribution and succession of fungi in the decaying CWD, using the denaturating gradient gel electophoresis (DGGE). To study the suitability of other, less toxic, density measuring techniques than the standard mercury one,samples were measured in a mercury medium as well as in a water medium. For water measurements we used Sartorius MSA 323S scale with YDK01 Density Determination Kit for the first time in our labs. We measured basic, absolute and saturated densities. Lower densities in higher decay phases were detected,as expected. The YDK01 Density Determination Kit proved to be a reliable method of measuring wood sample densities in water. However, some optimizations will have to be considered in the future. Molecular methods for studying fungi in CWD are in Slovenia still in early phases. Based on DNA extracted from fungi inside CWD, we conducted a DGGE analysis, resulting in fungi community profiles for each sample. According to the results, different decay stages host different types of fungi species. The number of fungi species per sample increases with higher decay phases. Genetic distances determined from DGGE profiles and geographic distances were not in high correlation.

S pridobitvijo Evropskega LIFE+ projekta LIFEGENMON je Gozdarski inštitut Slovenije skupaj s partnerji iz še dveh držav pridobil možnost za razvoj sistema gozdnega genetskega monitoringa (GGM) za dve drevesni vrsti – navadno bukev in belo jelko. V nalogi skušamo odgovoriti na zastavljeno hipotezo, ali so izvorne metode (protokoli) primerne za učinkovito spremljanje genetske pestrosti gozdov iz demografskega in fenološkega vidika? Predstavljene so metode in triletni rezultati dela na dveh ploskvah za GGM v Sloveniji. Po izboru ploskev, pri katerem smo upoštevali dopolnjene mednarodne kriterije, predstavljene v nalogi, smo na vsaki izmed ploskev vzorčili in označili 250 odraslih dreves in 200 dreves v pomladku. Na vsaki ploskvi smo zabeležili osnovne sestojne značilnosti, ocenili obrod, če je bilo to mogoče, in preverili gospodarjenje s sestojem znotraj ploskve v preteklosti. Na 40 izbranih odraslih drevesih in v pomladitvenih jedrih smo spremljali fenološke faze brstenja, moškega in ženskega cvetenja, pri bukvi tudi rumenenja listov. Zbrane podatke smo shranjevali v LIFEGENMON bazo podatkov, ki smo jo oblikovali posebej za ta projekt. Rezultati kažejo, da je vpeljava uniformnega koncepta GGM težavna zaradi različne biologije izbranih drevesnih vrst. Pri ocenjevanju fenologije opažamo majhne razlike v pričetku določenih fenofaz med opazovanimi leti. Predlagamo nekatere dopolnitve trenutnih protokolov za spremljanje genetske pestrosti gozdov. Slovenian Forestry Institute is developing a system for Forest Genetic Monitoring (FGM) for two species – Fagus sylvatica and Abies alba within the LIFE+ project LIFEGENMON. In this Thesis we test a hypothesis whether the protocols established at the beginning of the project meet the demands for efficient monitoring of genetic diversity from a demographic and phenological point of view. We present the methods used and results of the first three years of work on the research plots in Slovenia. After the plots were established, we sampled and marked 250 adult trees and 200 seedlings from natural regeneration on each of the plots. We assessed general stand properties, fructification and history of forest management. On 40 trees per plot and in the natural regeneration, phenological stages of bud bursting, male/female flowering and in beech also leaf senescence were assessed. The data was stored in a customly designed database. The results suggest difficulties in implementing an unified FGM system due to the different biology of species. We observed small differences between the start of different phenological stages between years. We propose some modifications of current protocols for their optimisation.

Od kakovosti in izvora gozdnega reprodukcijskega materiala (GRM) bodo odvisni genetska pestrost, struktura, preživetje in uspevanje sadik ter končno uspevanje in odpornost bodočega gozda. Zato mora Gozdarski inštitut Slovenije (GIS) na podlagi javnega pooblastila po Zakonu o gozdovih in zahtev zakonodaje o GRM preveriti njegov izvor pred izdajo glavnega spričevala ali na zahtevo inšpektorata kadarkoli v času trženja in uporabe. Uporaba molekularnih metod pripomore k vedenju o izvoru in genetski kakovosti GRM. GIS kontinuirano razvija znanje, infrastrukturo, gensko banko in molekularne baze podatkov za izvajanje opisanih testov. Zaradi suma o (načrtno) napačni navedbi izvora GRM, nabranega jeseni 2016 iz vsaj enega semenskega objekta, smo v predstavljeni študiji analizirali izvor in genetsko pestrost GRM iz štirih semenskih objektov bukve (Fagus sylvatica L.), v katerih je bilo seme nabrano v istem letu, ter drugih vzorcev iz Slovenske gozdne genske banke, skupaj petnajst. Za analize smo uporabili jedrne mikrosatelite, šestnajst lokusov, ki jih na GIS uporabljamo rutinsko. S tehničnega vidika so se izbrani markerji izkazali za primerne za identifikacijo posameznikov, oceno genetske pestrosti in identifikacijo domnevnega izvora. Zaradi možnosti mešanja vzorcev, pridobljenih s tal, in tehničnih potreb bomo v prihodnosti za potrebe rekonstrukcije genotipa semenskega drevesa analizirali vsaj dvanajst semen na drevo. Genetska pestrost manjših vzorcev je bila značilno manjša od tiste v velikih vzorcih. Opozarjamo, da je GRM nujno treba nabirati najmanj z v odobritvi semenskega objekta predpisanega števila dreves, da zagotovimo ustrezno veliko genetsko pestrost GRM, ki ga sadimo v gozdove. Metode razvrščanja posameznikov na podlagi Bayesove verjetnosti in filogenetska drevesa so pravilno določili izvor referenčnih vzorcev, medtem ko je bila resolucija analize glavnih komponent manjša. Vse metode, uporabljene za identifikacijo domnevnega izvora vzorcev semena, so nedvoumno pokazale, da vzorec TURs ni bil nabran v sestoju TUR, različen je tudi od preostalega analiziranega genofonda bukve v Sloveniji. Quality and origin of forest reproductive material (FRM) define the possible genetic diversity, structure, survival and development of seedlings and resilience of the future forest to stress and disturbances. The Slovenian Forestry Institute (SFI) must, based on the public authorization, in accordance to the Forest Act, The Act on Forest Reproductive Material and other legislation requirements concerning FRM, check its origin before issuing the master certificate or, on demand of the inspectorate, at any time during its marketing and use. Application of molecular methods contributes to the determination of FRM origin and its genetic quality. SFI continually develops know-how, infrastructure, gene bank, and molecular databases for performing such tests. In this study, the origin (provenance) and genetic diversity of FRM collected from four beech (Fagus sylvatica L.) seed stands in 2016 were analysed, resulting on the suspicion of (intentional) mislabelling of the origin of the FRM, and compared to samples stored in the Slovenian Forest Gene Bank. In total 15 samples were analysed using 16 nuclear microsatellite loci. From the technical point of view, the selected markers proved to be appropriate for individual tree identification, evaluation of genetic diversity, and identification of the alleged origin. Due to the possible mixing of samples, for which seed was collected from the ground, and technical needs, we will analyze in the future at least 12 seeds per tree for the needs of the seed tree genotype reconstruction. Genetic diversity of smaller samples (seed collected from less seed trees) was significantly lower than that of large ones. We would like to emphasize that FRM should be collected at least from the number of trees prescribed in the decree on approval of the seed object in order to safeguard genetic diversity of FRM. Methods of clustering using Bayesian methods and phylogenetic trees correctly determined the origin of reference samples, while the resolution of the principle component analysis was lower. All methods used for identification of the alleged origin of seed samples, unambiguously proved that TURs sample was not collected in TUR seed stand, and it also differed from the rest of the analysed beech gene pool in Slovenia. In the presented case study, the applied methods for the molecular identification of FRM origin proved to be necessary and prevented introduction of the inappropriate FRM into Slovenian forests.

Quality and origin of forest reproductive material (FRM) define the possible genetic diversity, structure, survival and development of seedlings and resilience of the future forest to stress and disturbances. The Slovenian Forestry Institute (SFI) must, based on the public authorization, in accordance to the Forest Act, The Act on Forest Reproductive Material and other legislation requirements concerning FRM, check its origin before issuing the master certificate or, on demand of the inspectorate, at any time during its marketing and use. Application of molecular methods contributes to the determination of FRM origin and its genetic quality. SFI continually develops know-how, infrastructure, gene bank, and molecular databases for performing such tests. In this study, the origin (provenance) and genetic diversity of FRM collected from four beech (Fagus sylvatica L.) seed stands in 2016 were analysed, resulting on the suspicion of (intentional) mislabelling of the origin of the FRM, and compared to samples stored in the Slovenian Forest Gene Bank. In total 15 samples were analysed using 16 nuclear microsatellite loci. From the technical point of view, the selected markers proved to be appropriate for individual tree identification, evaluation of genetic diversity, and identification of the alleged origin. Due to the possible mixing of samples, for which seed was collected from the ground, and technical needs, we will analyze in the future at least 12 seeds per tree for the needs of the seed tree genotype reconstruction. Genetic diversity of smaller samples (seed collected from less seed trees) was significantly lower than that of large ones. We would like to emphasize that FRM should be collected at least from the number of trees prescribed in the decree on approval of the seed object in order to safeguard genetic diversity of FRM. Methods of clustering using Bayesian methods and phylogenetic trees correctly determined the origin of reference samples, while the resolution of the principle component analysis was lower. All methods used for identification of the alleged origin of seed samples, unambiguously proved that TURs sample was not collected in TUR seed stand, and it also differed from the rest of the analysed beech gene pool in Slovenia. In the presented case study, the applied methods for the molecular identification of FRM origin proved to be necessary and prevented introduction of the inappropriate FRM into Slovenian forests. Od kakovosti in izvora gozdnega reprodukcijskega materiala (GRM) bodo odvisni genetska pestrost, struktura, preživetje in uspevanje sadik ter končno uspevanje in odpornost bodočega gozda. Zato mora Gozdarski inštitut Slovenije (GIS) na podlagi javnega pooblastila po Zakonu o gozdovih in zahtev zakonodaje o GRM preveriti njegov izvor pred izdajo glavnega spričevala ali na zahtevo inšpektorata kadarkoli v času trženja in uporabe. Uporaba molekularnih metod pripomore k vedenju o izvoru in genetski kakovosti GRM. GIS kontinuirano razvija znanje, infrastrukturo, gensko banko in molekularne baze podatkov za izvajanje opisanih testov. Zaradi suma o (načrtno) napačni navedbi izvora GRM, nabranega jeseni 2016 iz vsaj enega semenskega objekta, smo v predstavljeni študiji analizirali izvor in genetsko pestrost GRM iz štirih semenskih objektov bukve (Fagus sylvatica L.), v katerih je bilo seme nabrano v istem letu, ter drugih vzorcev iz Slovenske gozdne genske banke, skupaj petnajst. Za analize smo uporabili jedrne mikrosatelite, šestnajst lokusov, ki jih na GIS uporabljamo rutinsko. S tehničnega vidika so se izbrani markerji izkazali za primerne za identifikacijo posameznikov, oceno genetske pestrosti in identifikacijo domnevnega izvora. Zaradi možnosti mešanja vzorcev, pridobljenih s tal, in tehničnih potreb bomo v prihodnosti za potrebe rekonstrukcije genotipa semenskega drevesa analizirali vsaj dvanajst semen na drevo. Genetska pestrost manjših vzorcev je bila značilno manjša od tiste v velikih vzorcih. Opozarjamo, da je GRM nujno treba nabirati najmanj z v odobritvi semenskega objekta predpisanega števila dreves, da zagotovimo ustrezno veliko genetsko pestrost GRM, ki ga sadimo v gozdove. Metode razvrščanja posameznikov na podlagi Bayesove verjetnosti in filogenetska drevesa so pravilno določili izvor referenčnih vzorcev, medtem ko je bila resolucija analize glavnih komponent manjša. Vse metode, uporabljene za identifikacijo domnevnega izvora vzorcev semena, so nedvoumno pokazale, da vzorec TURs ni bil nabran v sestoju TUR, različen je tudi od preostalega analiziranega genofonda bukve v Sloveniji.

Measuring density of wood samples with mercury volumemeter is a standard throughout the world. To find out the reliability of another less toxic density measuring technique, samples were measured in a mercury medium and in a water medium as well. This study is a revision of the previously published results (Finžgar et al., 2013), since the determination of decay phases was questionable and correct results are presented here. New data is also presented herewith. For water measurements, the MSA 323S scale (Sartorius, Germany) with YDK01 Density Determination Kit (Sartorius, Germany) was used for the first time in our labs. We measured density of coarse woody debris from the secondary virgin forest reserve Rajhenavski Rog, Slovenia. Results of the two methods employed were not comparable due to different density definitions. Also, lower densities in higher decay phases were not detected, although expected. Nevertheless, the scale MSA 323S proved to be a reliable method of measuring wood sample density in water. However, some optimizations will have to be considered in the future. V Sloveniji in po svetu se je merjenje gostote v živosrebrnem mediju obravnavalo kot standardni postopek. Merjenje gostot v živem srebru je problematično zaradi zahtev Evropske unije po umiku te nevarne snovi iz človekovega okolja. Zato so meritve gostot v tej raziskavi potekale v različnih medijih z namenom iskanja alternativnega postopka. Ta raziskava revidira že objavljene rezultate (Finžgar in sod., 2013), saj je bila takratna določitev nekaterih stopenj razkroja vprašljiva. Prav tako so predstavljeni nekateri povsem novi rezultati. Tehtnica MSA 323S z dodatkom za merjenje gostot YDK01 (Sartorius, Nemčija) je bila v laboratoriju Gozdarskega inštituta Slovenije uporabljena prvič. Merili smo gostoto vzorcev velikih lesnih ostankov iz pragozdnega rezervata Rajhenavski Rog v vodnem in živosrebrnem mediju. Rezultati med seboj niso bili primerljivi, saj so bile uporabljene različne definicije gostote. Prav tako nismo zaznali izrazito manjših gostot v višjih stopnjah razkroja. Z našo raziskavo potrjujemo, da gostota lesa z razkrojem pada. Merilna tehtnica MSA 323S z dodatkom za merjenje gostot se je izkazala za izjemno hitro in preprosto metodo merjenja gostot lesa, čeprav bo treba v prihodnjih raziskavah postopek optimizirati.