The Forest Research Plot was chosen on a typical site for the Pokljuka plateau. It is covered by a supposedly autochthonous old Norway spruce stand with rejuvenation centres. Data on the physiological parameters can be used as representation values for a subalpine Norway spruce stand. The studies include: standard site and dendrological parameters, physiological parameters (foliar analysis, antioxidants, hormones and aminoacids in spruce needs), genetical and biochemical analysis (isoenzymes and molecular markers), microbiological activity (acid phosphatase, rate of decomposition, number and occurrence of fungal decomposers of wood and litter) and studies of symbionts and pathogens of Norway spruce (mapping of fruitbodies of mycorrhizal fungi, types of ectomycorrhizae, endophytes, epiphytes - lichens and phytopathological surveys). All physiological and ecological studies are planned to be repeated at certain time periods in order to understand better the functioning of the forest ecosystems on Pokljuka.

Mechanisms of air pollution toxicity are very complex and depend on various physiological and biochemical properties of plants. These mechanisms (including formation of free radicals) are still poorly understood. In addition, physiological responses of forest plants to air pollution stress can be modified by various biotic (e.g., insects, pathogens, mycorrhizae associations, genetic variation) and abiotic (e.g., increasing CO2 concentrations, ultraviolet-B radiation, nitrogen desposition, nutrient deficiencies, drought) factors. An example of air pollution effects on forest trees may be responses of ponderosa pine seedlings to elevated concentrations of ozone: various physiological changes in trees (lowered net photosynthesis, altered carbon allocation, deterioration of photosynthetic pigments, etc.) have led to reduced growth and biomass of plants.

Plant hormones play a major role in the development and control of a wide range of plant physiological processes. Here we examine the ways in which they may influence the response of plants to air pollutants both indirectly, for example through the control of stomatal behaviour by ABA and directly, through their reactions with oxidising pollutants, which can lead to the formation of further damaging molecular species.

The evaluation of data on the forest tree condition is weak point in forest decline monitoring as it enables only a very rough estimation of the state of forest health. The paper analyses the interpretation of the forest tree defoliation data in order to estimate their relevance and applicability. Quality control results of defoliation assessment in Slovenia are presented and the state of defoliation with its changes in the year 1987, 1991, 1993, 1994 on the 16 km grid.

We have been isolating AC/GT and AG/CT SSRs from the Norway spruce (Picea abies K.) nuclear genome. We isolated several hundreds positive clones from a small-insert genomic library and following sequence analysis we designed primers for 36 of them, 24 containing AG and 12 AC SSRs. After testing them on a panel of spruce individuals 25 of the primer pairs producted a single-locus hypervariable pattern, with the remaining ones giving either a single monomorphic product (18) or very poor amplification (19) or amplification of multiple bands (38). Segregation in accordance with a simple Mendelian model of inheritance was demonstrated for all the loci amplified with the primer pairs giving a simple variable pattern. We screened a panel of 19 spruce trees at these loci. The average number of alleles per locus was 14 and expected heterozygosity 0.80, with up to 23 alleles per locus and heterozygosities exceeding 0.94. This shows that nuclear SSRs can be very useful markers in the population genetics of trees even though the overall efficiency of the marker identification process is quite low due to the high percentage of primer pairs producting complex or "dirty" patterns. We attribute this phenomenon to the high complexity of the spruce genome. Other methods, including the construction of libraries highly enriched for SSR sequences, that we developed in order to make SSR retrival and typing easier and faster will be discussed. We recently extended the use of PCR amplified SSR markers to the chloroplast genome. We demonstrated that mononucleotide poly(A/T) stretches are frequent in the chloroplast genomes of plants and show high levels of between and within population variation, making them ideal tools for cytoplasmic population genetic overcoming the difficulties in finding within species variation that are frequently encountered when analysing the cpDNA molecule by RFLPs or PCR-RFLPs. We will present results of the analysis of mediterranean pine species populations by using a set of cpSSRs that are distributed over the whole cpDNA molecule and discuss the possible applications of such markers for studying gene flow and for paternity analysis.

Giemsa C-banding method was applied on chromosomes in the female gametophyte tissue of Norway spruce (Picea abies (L.) Karesten). A karyogram of one single tree growing in the Botanical Garden of the University of Graz, Austria, was made by using an image analysis system. All chromosomes (n=12) showed pericentic C-bands. Chromosomes II, V and IX possed intercalary C-bands located at secondary constrictions. Telomeric C-bands appeared in chromosomes II and III on both arms.

Cytogenetical and biochemical investigations of common oak (Quercus robur L.) were applied in order to find suitable markers in bioindication of oak populations. In cytogenetical analyses the chromosome aberration and mitotic index were recorded. Chromosome banding, B-chromosomes, location and number of nuclear organizer regions were indicated using Giemsa C-banding method, fluorescence staining and silver impregnation. In biochemical analyses SDS-electrophoresis of soluble proteins and isoelectric focusing of isoperoxidases were used.

It was our aim to investigate whether the cytokinin status of Norway spruce seedlings (Picea abies L. Karst.) was changed when the plants were grown under nutritive stress. Cytokinins in shoots and roots from spruce seedlings, grown in various hydrocultural media with complete (control) or poor nutrient supply (stress), were determined by an indirect competitive enzyme-liked immunosorbent assay. The shoots of spruce seedlings grown in a poor acidic culture medium, to which aluminium ions (AlCl3, 0.8 mM) were added, showed up to 5-fold higher concentrations of zeatin riboside and isopentenyladenosine (iPa). When adding naphtylacetic acid to the nutrient medium also higher levels of cytokinin ribosides were measured in the shoots. In the roots however cytokinin riboside levels did not increase significantly under Al- or auxin treatment. Feeding experiments using tritiated iPA have shown that the metabolism of this cytokinin is strongly retarded in the roots of stressed seedlings. The experiments show that acid conditions and nutrient shortage together with aluminium ions in the rhizosphere increase the number of lateral roots in Norway spruce seedlings. It is assumed that the increased number of sites of cytokinin biosynthesis and the reduced metabolism of iPA result in the higher content of cytokinin ribosides in the shoots of seedlings.

In 1992, a monitoring survey on a national scale was carried out using Hypogymnia physodes (L.) Nyl. as a biomonitor for trace element air pollution. The primary aim was to analyse epiphytic lichens collected at 86 sampling locations of the 16 x 16 km bioindication grid using k(0)-based instrumental neutron activation analysis (k(0)-INAA), and X-ray fluorescence spectrometry (XRF) for sulphur and lead, to obtain information about the levels of elements in the atmosphere and to identify significant pollution sources. The geographical concentration patterns of the trace elements obtained from the lichen data were mapped and compared with the index of atmospheric purity (IAP) calculated on the basis of data from lichen thalli type mapping, obtained on a more dense bioindication grid in 1991. The results obtained showed good agreement between the mapping of sulphur and trace elements with the status of lichen vegetation. The most exposed regions with elevated trace element levels and lower values of IAP were in the north-western Alpine part of Slovenia which coincides with high precipitation, and in the east of Slovenia, where many local pollution sources are situated.