Chloroplast DNA polymorphism of an exotic P. mugo Turra population introduced to seaside spit of Kursiu Nerija in Lithuania

The aim of this study was to elucidate the introduction history of P. mugo in the unique landscape of the Lithuanian seaside spit of Kursiu Nerija by assessing its genetic structure and the genetic diversity. The individuals were sampled in 12 populations within an area of 3 km × 50 km along the Lithuanian part of Kursiu Nerija. P. mugo was introduced over 200 years ago to prevent sand erosion by establishing a forest cover. Chloroplast DNA polymorphism of 220 individuals of P. mugo together with 18 P. sylvestris and 11 putative P. sylvestris × P. mugo hybrids was assessed by the aid of five microsatellite markers. The standard intra-population diversity indexes were calculated. The intra-specific variation between distinct morphotypes as well as the population differentiation within the most spread P. mugo ssp. rotundata morphotype was assessed based on the haplotype frequencies by hierarchical AMOVA, G_ST/R_ST test, UPGMA clustering and PCA methods. The genetic diversity of P. mugo in Kursiu Nerija was high (H_e = 0.95; 83 different haplotypes). All except one of the P. mugo populations sampled contained a notable share of private haplotypes. AMOVA revealed high intra-specific diversity but low differentiation between the P. mugo populations. Most of the haplotypic variance was within populations. The UPGMA clustering produced groups more corresponding to the sub-species morphotypes than the geography of the populations. There was no geographical pattern of reduction in genetic diversity towards the younger plantations. A strong candidate for a species-specific DNA marker was found. After several events of introduction, the genetic diversity of P. mugo in Kursiu Nerija is very high and is structured based on the sub-species morphotypes rather than geography. The high frequency of shared and notable frequency of private haplotypes in most of the populations indicate that the major part of the P. mugo material originates from a number of geographically and genetically related sources, which more likely are introductions from abroad that the local collections. The high frequency of private haplotypes in the northernmost populations leaves a possibility for minor introductions from other genetically distinct sources. The absence of private haplotypes in one of the sampled populations indicates the use of local seed collections. The large number of shared haplotypes provides a strong evidence for a geneflow among the P. mugo taxa.     

Reduction of herbivore density as a tool for reduction of herbivore browsing on palatable tree species

Browsing of overabundant free-living herbivores often limits the natural as well as artificial regeneration of forest in Central-European Mountains. The aim of the study was to assess the efficiency of the extensive reductions of herbivore populations for protection of forest regeneration in preferred areas. We analysed the relationship between the intensity of shoot browsing and relative density of herbivores in viewpoint of natural regeneration of secondary mountain spruce stands with low proportion of broadleaved trees in Králicky Sněžník Reserve (Czech Republic). The distribution of seasonally migrating herbivores was established by counting faecal pellet groups (standing crop method) in growing season. The intensity of browsing of spruce, beech and rowan was assessed using the proportion of browsed shoots on individual tree sapling. Density of red deer was re-counted from the number of pellet groups and it was 15–56 individual/km². The intensity of spruce browsing was low in the whole area in all seasons. In general, the intensity of rowan and beech shoot browsing was high in the whole area. There were no correlations between deer density and browsing intensity of rowan (browsing was severe in all plots). We conclude that the natural regeneration of attractive trees and shrubs is nearly impossible in habitats where proportion of these food items is too small, even when the density of herbivores is low. Reduction of herbivores density is useful for protecting spruce and other tree species with low preference by herbivores. Forest managers should take into account the high palatability of deciduous tree species and distribution of herbivores on localities. Preference of localities is often based on grasses in the food supply. Regeneration of highly palatable tree species requires both low density of herbivores and sufficient protection of saplings.     

Effects of drought and rewatering on growth and transpiration in European beech seedlings late in the growing season

Drought stress is one of the most important environmental factors affecting plant growth and survival. To date, most studies aim at understanding of post-stress physiological and anatomical adaptation to drought stress; however only few studies focus on plant recovery. In the present study, transpiration, shoot water potential, and anatomical and morphological measurements were performed on 4-year-old European beech seedlings with fully developed leaves. The seedlings were exposed to three levels of soil water potential (well-watered, moderate drought stress and severe drought stress) and followed by rewatering under greenhouse conditions. Reduced transpiration rates were observed in the stressed seedlings as a response to drought stress, whereas anatomical and morphological variables remained unchanged. Three days after rewatering, transpiration rates in both moderately and severely stressed seedlings recovered to the levels of those of well-watered seedlings. Drought stress promoted leaf budding, resulting in higher shoot dry mass of stressed seedlings. Our findings indicate that anatomical and morphological adaptations of European beech seedlings to drought stress are visibly limited during late-season growth stages. These results will help us to further understand factors involved in drought adaptation potential of European beech seedlings faced with expected climate-related environmental changes. To complete our findings, further experiments on plant recovery from drought stress should be focused on different periods of growing season.     

Needle blight caused by Dothistroma pini in Slovakia: distribution,host range and mating types

Dothistroma needle blight (DNB) has been observed in Slovakia during the last two decades. Up until 2017, Dothistroma septosporum has only been detected and molecularly confirmed to cause DNB in Slovakia. Here, we report the detection of Dothistroma pini at six localities around Slovakia, representing different plantation types. Four pine species (Pinus sylvestris, P. nigra, P. mugo and P. jeffreyi) were confirmed as hosts of D. pini in Slovakia, of which only P. mugo has been previously reported as host in Slovakia. Three gene regions (ITS, EF1 –α, and ß-tubulin) of each of the 13 isolates were sequenced and assigned as D. pini. Based on ITS sequences, the studied isolates represent the haplotypes Dp_HAP.1, Dp_HAP.2. Both mating types were detected but at different localities. Our results suggest that in addition to D. septosporum, D. pini may contribute to DNB also in Slovakia.     

Occurrence and pathogenicity of Phytophthora × cambivora on Prunus laurocerasus in Serbia

Cherry laurel (Prunus laurocerasus) is a native plant species in Serbian forests, but is also widely used for ornamental plantings. Following two extremely wet summers in 2014 and 2015, in spring and summer of 2016 and 2017, numerous cherry laurel plants with symptoms indicative for Phytophthora diseases, like wilting and chlorosis of leaves, dieback and bleeding bark necroses, were recorded in a park in Belgrade and in two ornamental nurseries in central Serbia. From necrotic bark samples and rhizosphere soil, self‐sterile Phytophthora isolates with woolly colonies were obtained. Due to the production of ellipsoid and elongated, non‐papillate sporangia in water and of ornamented oogonia with two‐celled antheridia in mating tests with tester strains of both Phytophthora × cambivora and P. cryptogea, these isolates were identified as P. ×cambivora which was confirmed by ITS sequence analysis. Pathogenicity of P. ×cambivora from cherry laurel (PCCL) was tested by inoculating one‐year‐old seedlings of cherry laurel under the bark. P. ×cambivora from European beech (PCB), and isolates of P. cactorum (CAC), P. cryptogea (CRY), P. plurivora (PLU) and P. ×serendipita (SER) were included as comparison. Three and a half months after inoculation, nine of the twelve plants in PCB, three in PCCL and CAC and two in PLU declined with longitudinal necroses and chlorosis, wilting and premature shedding of leaves. These results demonstrate the ability of P. ×cambivora to infect and cause decline of cherry laurel plants. The particularly high aggressiveness of the P. ×cambivora isolate from beech shows that this pathogen poses a serious risk to cherry laurel in the rare natural communities of cherry laurel and beech in Serbia.     

Landscape effect for the Cervidaes Cervidae in human-dominated fragmented forests

We analyse the relationships between the main Cervidae [moose (Alces alces), red deer (Cervus elaphus) and roe deer (Capreolus capreolus)] species and a complex of environmental factors in an extensive fragmented landscape of Central Lithuania. The highest determining positive influence on moose density was the proportions of wet forest sites. In forest complexes with fewer proportions of wet sites, the most important factor was the total forest area. The proportion of shrub cover, upland and dense undergrowth area, and road density also has significant effect on moose density. The total area of forest complexes has the highest determining positive influence on red deer density. The highest density of red deer was calculated in large forest complexes (>2,745 ha) with a <17.6 % proportion of pine and <36.5 % of deciduous forests. Other significant factors were core area, road density and urbanization level. Forest edge density has the highest influence on the roe deer populations. The highest density of roe deer was recorded on forest areas with >51 m ha^?1 of edges in wet forest (>25.4 %) dominating areas. The proportion of deciduous, coniferous mixed and pine forest, also shrub and density of edges also had significant effect.     

Distribution of Aeolothrips intermedius Bagnall (Thysanoptera: Aeolothripidae) and its potential prey Thysanoptera species on different cultivated host plants

The results of the monitoring of Thysanoptera species on cultivated plants in Slovenia (2000–2001), Croatia (1994–1996), and Serbia and Montenegro (1988–2003) are presented in this study. The aim of the investigation was to study the host plant distribution of the predator Aeolothrips intermedius Bagnall and its potential prey. Banded thrips were found on 30 different host plant species belonging to 16 botanical families, always in mixed populations with phytophagous or facultative phytophagous insects (including 18 Thysanoptera species). On the vegetative parts of the cultivated plants, banded thrips were found less numerous in spite of the massive population of some harmful thrips species. This indicates highly important role of pollen as alternative food for Aeolothrips intermedius.     

Manure application followed by biochar application increases plant production regardless of soil dehydrogenase activity

Biochar is obtained by the pyrolysis of biomass, and contains abundant carbon and minerals. Biochar supplementation of soils can greatly improve soil health and quality, but these beneficial effects typically develop slowly over time. Depending on the quality of the biochar and the soil to which it is applied, it may take years before positive effects are apparent. This is because organic substances are slowly sorbed onto the biochar over time, and the biochar eventually becomes part of the sorption complex of the soil. It is therefore advisable to apply biochar together with some organic material. We examined the effect of co-application of different doses of biochar with manure on soil dehydrogenase activity (DHA), soil oxidizable carbon (COX), cumulative soil respiration, soil buffering capacity, the soil exchange reaction (pH/KCl) and the production yield of winter rape seeds. We also determined seed production when artificial granular fertilizers were added to biochar and manure. The results showed that the application of biochar and manure significantly increased grain yield, DHA, the soil exchange reaction and cumulative respiration. Thus, application of biochar with organic material can increase seed yield and some properties of agricultural soils. However, the positive effect of biochar on seed yield was not directly proportional to biochar dose, in that the seed yield was lower for a biochar dose of 45 t/ha than 30 t/ha.     

Bioindication of Different Stresses in Forest Decline Studies in Slovenia

Determination of the stresses operating on a forest ecosystem demands the use of several bioindication methods. Air quality indicators were assessed from an inventory of forest decline based on the assessment of tree crowns and epiphytic lichens. Photosynthetic pigments, ascorbic acid and major macronutrients were studied in Norway spruce needles as indicators of physiological and biochemical stress. Analyses were carried out on selected forest plots in polluted areas (Zasavje district, vicinity of thermal power plants) and predominantly unpolluted areas (Triglav National Park, Julian Alps, Slovenia). For some bioindication methods, there was good agreement with measured air quality and climatological parameters. The best agreement was found between total foliar sulphur in needles and epiphytic lichens, especially in more polluted areas. Agreement with forest decline inventories and analyses of some needle stress physiological/biochemical parameters was less convincing. The strength of agreement was further decreased by soil characteristics and climatic parameters, influenced also by biotic parameters and forest stand history and management. It was concluded that there is no simple bioindication method available to evaluate the vitality of a forest.     

Transformation of Petroleum Saturated Hydrocarbons during Soil Bioremediation Experiments

This paper presents transformations of saturated hydrocarbons of petroleum type pollutants during ex situ bioremediation of soil on the pilot heap (halde), during a period of 6 months, within the grounds of Petroleum Refinery Pan?evo (Serbia). Samples for analysis were taken in time intervals of 2 weeks (P₁–P₁₂ samples). Organic substance was extracted by Soxhlet’s method and quantified. Isoprenoid aliphatics, in particular pristane and phytane, and polycyclic aliphatics of sterane and triterpane types in saturated hydrocarbon fractions were analysed by GC-MS (SIM method). Significant amounts of n-alkanes have not been detected. The MS-chromatogram revealed only marginal amounts of pristane and phytane in sample P₁. Pristane and phytane occurred in sample P₈, and in even higher quantities in the final sample P_12. The proceeding bioremediation process was accompanied by the decrease of the relative amounts of pentacyclic terpanes of hopane type, compared to tri- and tetracyclic terpanes. In the initial sample P₁ the distribution of steranes and hopanes follows a pattern, which is characteristic for crude oils. However, their identification by SIM method was not possible in samples P₈ and P₁₂ because of the reduced concentration. The observed changes in the alkane fractions’ compositions may be considered as atypical, referring to the fact that during oil biodegradation under natural conditions, decomposition of isoprenoids occurs much easier and faster than decomposition of polycyclic alkanes of tri-, tetra- and pentacyclic terpane, sterane and diasterane types, after the decomposition of n-alkanes has been almost completed.