In ecological research, quantitative methods are often used to measure the total content of metabolites groups (i.e., phenols, carbohydrates). Until recently, there has been no simple and effective method to determine the total terpenoid content with satisfactory repeatability and sensitivity. The procedure proposed by Ghorai et al. (2012) requires the use of fresh plant material. That may be problematic when experimental units are located far from a laboratory. Our goals were to optimize the procedure, and to find the threshold of misestimation using the procedure adjusted to work with dried material. Needles of Pinus sylvestris were used to test the effect of changes in drying, grinding, storage, and extraction on determined total terpenoid content. All applied changes in material storage and grinding decreased the quantity of the terpenoids in needles. Only air-dried and ball-milled material produced similar results to those obtained with fresh material - can be recommended if the fresh material unattainable. Air-dried material may be stored for up to three months, but it resulted in greater variation and then greater sample size is needed. Lower sample mass and solvent volume have no impact on accuracy. Shorter extraction time, oven-drying or microwaving leads to unreliability of measurements.

Woody plants growing along streets and construction sites play an important role in removing harmful particulate matter (PM). Researchers rarely consider the impact of different types and size fractions of PM deposited on the leaves on insect folivores. We determined differences in the accumulation of cement and roadside PM on the leaves of two Prunus species (P. padus and P. serotina) with different leaf surface structures. We also determined the effect of PM on the beetle Gonioctena quinquepunctata, the main pest of these plants. Saplings were artificially dusted in greenhouses and leaves were utilised for larval and adult insect stages feeding in laboratory conditions. Road PM accumulated in greater amounts than did cement PM, regardless of plant species. For both PM sources, P. padus accumulated twofold more than did P. serotina. Insect survival was negatively affected by PM pollution; however, neither Prunus species nor PM source variant significantly affected masses of larvae and pupae, duration of larval and pupal development or relative growth rates. The experiment showed strong negative influences of PM were noted only for adult insects, due to the grazing period being longer than that in larvae. The mass of adult insects and the efficiency of conversion of ingested food (ECI) were lower for insects exposed to PM than those for control insects. Insects compensated for lower ECI by eating a greater total amount of food (TFE). Adult insects gained significantly higher mass when fed with P. serotina than with P. padus. The effect of PM on analysed plant metabolites was insignificant. Only Prunus sp. and date of collection affected the level of condensed tannins and total phenols. Our results indicate that, when investigating the effect of the host plant on folivore performance, the accumulation of PM, as well as its type and quantity, should be taken into account.

Defoliation caused annually by Cameraria ohridella has a considerable impact on the general condition of horse chestnut trees. We investigated the consequences of tree endotherapy against this insect. Wood samples, leaves and insect specimens were collected ten years after the treatment (injection) with pesticides. Insect invasion was manifested in all investigated trees by distinct radial growth depression. Treatment resulted in a growth upturn, while non-treated trees maintained decreased growth. The overall foliage infestation level was lower in treated trees, but pupal mass and potential fecundity of insect females were not affected. Our study revealed a long-lasting, positive effect of endotherapy on trees’ appearance and growth.

Instytut Dendrologii PAN w Kórniku - Institute of Dendrology PAS (Kórnik, Poland)

Horse chestnut leaf miner (Cameraria ohridella) has achieved ecological success by colonizing the entire European range of its primary host, horse chestnut (Aesculus hippocastanum). This insect has attracted the attention of scientists, but its ecology is poorly understood. Here, we investigated the effects of varying degrees of light availability on the leaf morphology of horse chestnut saplings and the performance of C. ohridella. A pot experiment under greenhouse conditions was performed in which the photosynthetic photon flux density (PPFD) was reduced from full light by 50% (high light - HL) or 80% (low light - LL). Insect performance parameters were quantified (i.e., pupal mass, efficiency of conversion of utilised leaf tissue (ECU) and potential fecundity). Compared with HL leaflets those from LL were characterised by higher contents of nitrogen and water but lower total phenolics. The oxidative capacity of phenolics (at pH ≈ 10, common in the lepidopteran gut) was low and did not differ in the two treatments. Compared with those collected from HL leaves, the mines of those collected from leaves of plants grown under LL conditions were larger in area but the leaf mass utilized by larvae was similar. Pupae were heavier in LL than in HL conditions, and ECU was higher in LL. The potential fecundity of females was not sensitive for experimental treatment. We conclude that (1) reduced light had a strong beneficial effect on the performance of C. ohridella and (2) phenolics in A. hippocastanum leaf tissues are a poor defence against this herbivore.

Rhizosphere microbial communities can influence plant growth and development. Natural regeneration processes take place in the tree stands of protected areas, which makes it possible to observe the natural changes taking place in the rhizosphere along with the development of the plants. This study aimed to determine the diversity (taxonomic and functional) of the rhizosphere fungal communities of Norway spruce growing in one of four developmental stages. Our research was based on the ITS region using Illumina system sequencing. Saprotrophs dominated in the studied rhizospheres, but their percentage share decreased with the age of the development group (for 51.91 from 43.13%). However, in the case of mycorrhizal fungi, an opposite trend was observed (16.96–26.75%). The most numerous genera were: saprotrophic Aspergillus (2.54–3.83%), Penicillium (6.47–12.86%), Pyrenochaeta (1.39–11.78%), pathogenic Curvularia (0.53–4.39%), and mycorrhizal Cortinarius (1.80–5.46%), Pseudotomentella (2.94–5.64%) and Tomentella (4.54–15.94%). The species composition of rhizosphere fungal communities was favorable for the regeneration of natural spruce and the development of multi-generational Norway spruce stands. The ratio of the abundance of saprotrophic and mycorrhizal fungi to the abundance of pathogens was high and promising for the durability of the large proportion of spruce in the Wigry National Park and for forest ecosystems in general.