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Multiplex detection of<i>Phytophthora</i>spp. using the Fluidigm platform

AbstractThe genusPhytophthoraplays an important role not only in agriculture but also in forest ecosystems. As the number of knownPhytophthoraspecies continues to grow, identifying new isolates in this genus has become increasingly challenging even by DNA sequencing. Therefore, the development of proper techniques for detection and identification is crucial for monitoring and control of these pathogens in the forestry sector. In recent years, new molecular methods using innovative approaches have indeed been developed. However, the majority of these methods was designed to detect singlePhytophthoraspecies. Techniques that are able to target multiple species would offer advantages, especially for the assessment of Phytophthora diversity in the environment. This paper describes a multiplex assay for the identification of eightPhytophthoraisolates, down to the species level, based on a Fluidigm platform employing pyrosequencing. The obtained results showed that for an accurate determination of the species, it is sufficient to know the sequence of two markers, ITS and COX1. The sensitivity of this test is sufficient to identifyPhytophthorain a pure culture. Unfortunately, analysis based on a pyrosequencing platform does not provide enough data to simultaneous identify multiplePhytophthoraspecies in samples collected in the field. This problem could be resolved in the future by sequencing using more efficient platforms like Illumina or IonTorrent.

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Recreation in nature reserves – preferences and satisfaction of tourists visiting the Polesie National Park

Abstract The aim of this work was to determine preferences and satisfaction of tourists visiting the Polesie National Park (PPN). Preferences were defined in terms of the motives for visiting, ways of spending leisure time, the length and frequency of visits, knowledge of tourist attractions and awareness of the Park’s financing. A survey was conducted in July–August 2019 gathering responses from 125 adults visiting the PPN tourist and bicycle paths. 100 correctly completed questionnaires were analysed using the CART method to determine the respondents’satisfaction with spending leisure time in the Park. The most frequently mentioned reasons for visiting were the beautiful landscape (28%) and the species richness (27%) of the PPN. 39% of respondents visited the area for the first time and 47% came for one day. Most visitors (65%) had very good knowledge of the tourist attractions in the PPN. Walking was the most common way (37%) for visitors to spend theirleisure time in the Park. More than half of the respondents (58%) would be willing to accept additional fees in order to help maintain and protect the PPN. The vast majority of the respondents indicated that they are satisfied (42%) or very satisfied (48%) with their visit to this area. Our statistical analysis indicated that asking the question about financing the Park greatly impacted the responses to the question about visitor satisfaction, but was also correlated with the respondent’s place of residence as well as their knowledge of tourist attractions. The unique character, landscape as well as the natural, historical and cultural richness of the PPN combined with the well-maintained infrastructure are crucial to ensure a high level of visitor satisfaction.

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Quality of Scots pine, European beech and pedunculate oak grown from sowing on soil with different compaction levels

Abstract In this study, we explore the effect of soil compaction on the growth of seedlings of Scots pine Pinus sylvestris L., European beech Fagus sylvatica L. and pedunculate oak Quercus robur L. On the experimental plots, ground contact pressures ranging from 0 to 250 kPa was applied on the soil. The applied pressure resulted in an increase in soil compaction between 1.02 to 1.19 g cm–3, which reflected pressures exerted by the undercarriage of vehicles used in logging. We then measured the seedlings as well as the dry weight of the roots and the above-ground parts. Using this data, we calculated the following quality indicators for each seedling: SQ – sturdiness quotient, S/R – shoot to root dry mass, DQI – Dickson quality index. For pedunculate oak, the SQ value significantly improved with increasing soil compaction, whereas no differences in the other two indicators were observed. In case of the European beech, the best value of SQ and DQI were observed at a soil density of 1.11 g cm–3, whilst no significant difference for the S/R coefficient could be found. Completely different results were obtained for Scots pine. The most favorable growth was observed when no pressure was applied. However, the SQ and S/R ratios even exceeded the values commonly considered acceptable. Our results therefore indicate that the values of seedling quality indicators are indeed influenced by soil compaction. At a soil compaction of 1.11 g cm–3, the share of seedlings with the SQ value below the critical level was the highest, but a similar relationship could not be confirmed for the other indicators. The response of the seedlings to compaction is likely to be species specific.

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Decline of Black Alder <i>Alnus glutinosa</i> (L.) Gaertn. along the Narewka River in the Białowieża Forest District

Abstract Black Alder Alnus glutinosa (L.) Gaertn. is an important tree commonly growing in Poland. Alders are actinorhizal plants that play an important ecological role in riparian ecosystems through atmospheric nitrogen fixation, filtration and purification of waterlogged soils as well as providing a refuge for terrestrial and aquatic organisms thus helping to stabilize stream banks. Black alder used to be considered a very pest and disease resistant species but, the situation changed in 2000, when an unprecedented decline of Alders was observed in Poland. In the Białowieża Forest District, this decline has been observed on wet meadow habitats and along rivers or watercourses. Currently, there are several hypotheses explaining Alder dieback, among them climatic changes and Phytophthora infections. In terms of climate, Black Alder requires a high atmospheric humidity during all phases of its reproductive cycle. It tolerates neither long-term summer flooding nor a significant decrease in the groundwater level. In terms of pests, oomycete pathogens of the genus Phytophthora are the most destructive plant pathogens known and many of them are present in forests and nurseries all over Europe. The aim of this study was to evaluate the health of Black Alder along the Narewka River in the Białowieża Forest District. Selected areas were monitored in 2012 and 2018, but no relationship between drought and alder health was found. A preliminary analysis of soil and water samples by real time PCR revealed the presence of two Phytophthora species: P. alni and P. cactorum. Further and more detailed research is required to elucidate the role of these pathogens in Alder dieback.

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The influence of stand and habitat characteristics on the occurrence of pine sawflies Diprion pini L. and Gilpinia virens (Klug) (Hymenoptera, Diprionidae) in selected areas of northern Poland

Abstract Pine sawflies (Hymenoptera: Diprionidae) belong to the most common pests of Scots pine Pinus sylvestris L. in the area of the Regional Directorates of State Forests in Toruń and Gdańsk. The two most destructive species, Diprion pini L. in the area of Puszcza Bydgoska and Gilpinia virens (Klug) in Bory Tucholskie, were investigated in this study. Our main aim was to characterise the two species in the Bydgoszcz Forest District in RDSF in Toruń and Kaliska in RDSF in Gdańsk, respectively, based on data gathered during outbreaks between 1991 and 2011. For this purpose, a geometric database was created, containing information about the year of outbreak, number of larvae observed after tree felling as well as selected stand and site features including stand age and species composition, stocking index, canopy closure, site index, humidity and fertility. By analysing the occurrence of both species in relation to stand and site features, we were able to determine their environmental requirements, which are quite different from one another. D. pini preferred older stands, with a stocking index between 0.8–0.9, growing on fresh and dry coniferous sites, while species composition (share of pine), canopy closure or site index did not have any influence on its abundance. G. virens preferred middle-aged pure pine stands, with relatively open canopies, on fresh coniferous sites and poorer soils. The stocking index did not affect its occurrence.

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The impact of fertilisers and fungicides on seed germination and the initial phase of seedling growth in black alder Alnus glutinosa (Gaertn.)

Abstract To test the influence of selected fertilisers and fungicides on the germination of black alder seeds and the initial phase of seedling growth, we conducted a laboratory experiment outlined in this paper. Six treatments were applied on petri dishes each containing 30 seeds. The substrate for germination was sterile filter paper wetted with an aqueous solution of either one of two fungicides, two organic fertilisers, a mineral fertiliser or distilled water (control). Fungicides and fertilisers were applied according to the manufacturers' recommendations. In order to keep genetic variability to a minimum, seeds originated from a single tree in a seed stand located in the Chotyłów Forest District, eastern Poland. Germination and growth took place at a temperature of 23°C ± 2°C with a 14 h/10 h day/night cycle. Seeds began to germinate as early as the second day after sowing, except for the mineral fertiliser treatment, in which the first sprouting was observed on day 3. Seedling length was measured daily from the day of germination of a given seed through to day 15. Germination was found to proceed most rapidly in the control, while the largest increments in length and dry mass occurred in the control and fertiliser treatment with the so-called N1 fertiliser (solely comprising growth stimulators in the form of humic acids, chitosan and silicon). The most limited growth was observed under the influence of the F1 fungicide (active compound Thiram) as well as the organic fertiliser N2 (a mixture of mineral components and organic growth stimulators). Roots were found to develop most rapidly in the control and in the treatment with N1 (no mineral components). These are also the only two treatments in which the roots were longer than the stems after 15 days. Fertiliser N2 was found to have the most unfavourable influence on both, germination and the first phase of seedling development. The fact that selected fertilizers and fungicides affected black alder seeds and seedlings under laboratory conditions does not mean that they will have an impact under field conditions or on other forest tree species. Therefore, this type of research will need to be conducted individually for each forest tree species.

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Fungi associated with<i>Armillaria</i>spp. rhizomorphs on<i>Populus tremula</i>L.

AbstractThe economic significance of aspenPopulus tremulal. is only marginal, which is the main reason for its low share in the structure of stands. however, aspen can play a decisive ecological role as a pioneer tree in the succession of forest communities and may decrease the occurrence of pathogenic fungi.Aspen is also dying out and in order to determine possible biotic causes, we examined taxa of fungi using classical and molecular methods in the area of the Czerwony Dwór Forest District, Subdistrict Rogale, compartments 74b and 75j. The health of aspen trees and the quantitative and qualitative structure of microorganisms associated withArmillariaspp. was evaluated.Based on our health assessment of the aspen, we conclude that the main reason for its dying out is the intensification of root rot, which is a result ofArmillariaspp weakening the trees. In the general pool ofArmillaria-associated microorganisms, saprotrophs were dominating, and the fungusFomitopsis pinicolacommonly inhabited the poplar stems, whereas pathogen antagonists comprised the lowest proportion. In the quantitative and qualitative structure of fungi found in rhizomorphs, the composition of ecological groups inhabiting stumps was found to be favourable nevertheless. The increasing frequency of pathogens on the logs is a major premise for removing trees affected byArmillaria, because aspen stumps are a good food base forA. cepistipesandA. ostoyaewhich ultimately spread within the stand. Treatment of stumps with preparations stimulating the decomposition of wood is not recommended for logs that form the food base for pathogenic fungi.

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Geocaching in education – a review of international experiences Part 3.Organisation of classes

AbstractThis article discusses the organization and conduct of educational activities using geocaching. In the scientific literature, the organization of geocaching classes is divided into three main stages: preparation, field work and evaluation. The preparation stage includes issues such as the role of the educator, the scenario itself, the duration of the classes, the length and course of the route, the number and location of caches, the coordinates and type of caches, potential descriptions of caches or the preparation of hints as well as the content of caches, necessary aids and preparing the students. In terms of the implementation of the fieldwork, the following issues need to be addressed: the role of the educator, introduction to classes, division of participants into teams, locating and working with caches as well as a summary of the activities. The evaluation stage should include an evaluation of the participants’ performance, the activities/thematic trails by the participants and the educational effect by the organizers. Additionally, other educational possibilities for using geocaching, such as tasks based on internet geocaching services, the use of existing caches and the establishment of caches by students are also briefly discussed.Due to the universality of the recommendations presented in the literature, educational geocaching can be implemented in every country, including Poland, both in formal and informal education, e.g. in the State Forests, national parks and landscape parks. Although this innovative teaching method involves a lot of initial work, its positive educational and social effects more than compensate for the invested time.

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