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In forestry management, artificially produced planting material is mainly used for renewal the tree population. Seedlings are cultivated in two systems: in the ground (the bare−root seedlings) and in controlled conditions (container seedlings). The aim of the study was to analyse the microsatellite markers of nuclear and chloroplast DNA, in terms of the number and frequency of rare, private, low frequency and common alleles in the planting material of Scots pine. The rare alleles included alleles occurring with less than 1% in analyzed group of seedlings and low frequency alleles occurred with a frequency of less than 25%. The private alleles were detected only in one group of seedlings. Genetic pools of seedlings from traditional (soil) and container production were compared. Planting material came from nurseries in the Olsztynek (N Poland) and the Oleszyce (S Poland) forest district. With the similar number of observed nDNA and cpDNA alleles in both analyzed locations, a higher number of rare, low frequency and private alleles was found within container seedlings. Most private alleles were a rare allele. Rare and private alleles are supposed to be responsible for adaptation to changing climatic conditions and a stressful environment. It seems reasonable to continue research on the meaning of rare and private alleles under conditions of strong selective pressure.
The production of forest tree species in forest nurseries is performed via two main breeding systems: i) the traditional (conventional) way with the seedlings grown in soil, and ii) plants cultivated in the containers. The aim of the study was to assess the level of genetic variability in the populations of the mother stands and the progeny populations of Scots pine cultured with traditional way (in soil) and in containers in two nurseries in Olsztynek (N Poland) and Oleszyce (S Poland) forest districts. Four polymorphic microsatellite markers (SPAG 7.14, SPAC 11.6, SPAC 12.5 and SsrPt_ctg4363) were used to evaluate the genetic variability of the studied populations. The basic hypothesis assumed that higher gene pool characterizes the seedlings grown in the containers comparing to the seedlings grown in the ground. The results confirmed that. Seedlings from containerized breeding had larger gene pool and were more diverse than plants with conventional breeding, both in Olsztynek and Oleszyce. Our study revealed a significant human impact on shaping the pool of forest genetic resources of Polish forests at the early stage of nursery production and showed the need for a broader study on further stages of cultivation of forests.
Swarming bat activity was monitored at three caves at elevations ranging from 880 m to 1,907 m above sea level in the Carpathian Mountains, using an infrared light barrier with data-logger, a video camera with a night-scope system, and subsequently by mist netting. A total of 6,175 bats of 19 species was captured, and over 70,000 passes through cave openings were registered. Caves differed in bat species richness, sex ratio, abundance of particular species and species composition. Peak species richness was observed in the mid-elevation cave. Bat activity was high in all caves, but declined with increasing altitude. Swarming activity occurred earlier at high elevation than at lower elevations. Activity of boreal-alpine species, such as Eptesicus nilssonii, peaked at the start of the swarming period, that of species typical of lower elevations, such as Myotis emarginatus, peaked in the middle of the swarming season. In a few species, males showed a significant preference for higher altitude caves, in contrast to females. A similar pattern was observed in the proportion of adults to juveniles, which increased with increasing elevation. Our results also suggest that M. brandtii and M. alcathoe were more often encountered at lower elevations, M. mystacinus (sensu stricto) at higher ones.
Traditional detection methods such as baiting or direct isolation take a long time and are incapable to handling large volume of material to be tested. The real−time PCR−based techniques are faster, more sensitive, more easily automated, and do not require post−amplification procedures. Species−specific primers for Phytophthora were designed based on the internal transcribed spacer regions (ITS) of rDNA collected from the NCBI DNA database. Primers and probes were designed using the Allele ID 7 at default search criteria. Specific probes were labeled with the reporter dyes JOE (6−carboxy−4,5−dichloro−2,7−dimethoxyfluorescein) at the 5' end and HBQ1 quencher at the 3' end (Sigma−Aldrich). The specificity of primers and fluorogenic probes was tested against genomic DNA of P. alni subsp. multiformis, P. lacustris and P. taxon hungarica. The real−time PCR reactions with the specific probes and primers yielded positive results with five concentrations of standards obtained by standard PCR reaction for corresponding Phytophthora species. The negative control (lack of DNA pathogens) yielded no amplification products. Standard curves showed a linear correlation between input DNA and cycle threshold (Ct) values with R² from 0.994 (P. alni) to 0.998 (P. taxon hungarica). The amplification efficiency of target DNA varied from 94.6% (P. alni) to 100% (P. taxon hungarica). The validation of the primers and probes designed for analysed Phytophthora species was performed on pure cultures, on soil samples from the forest nursery and declining oak stands. The designed probes displayed the high specificity of the detection of investigated species in pure cultures. The presented new molecular TaqMan probes can fully assist the integrated pest management as a powerful tool for a quick detection of above pathogenic organisms in forest nurseries. The molecular detection of harmful phytophthoras and in consequences diminishing of fungicides use for their control in forestry fully support European Union directives as well as the ‘Good plant protection practice measures' elaborated by European and Mediterranean Organisation of Plant Protection.
The study was conducted in 2015 in six spruce stands situated in different forest districts administratively belonging to the Regional Directorate of Forests State in Krosno. Each spruce population was represented by 30 trees and assessed in terms of their current health status. Genetic analyses were performed based on shoot samples from each tree using nine nuclear DNA markers and one mitochondrial DNA marker (nad1). The health status of the trees was described according to the classification developed by Szczepkowski and Tarasiuk (2005) and the correlation between health classes and the level of genetic variability was computed with STATISTICA (α = 0.05). Nuclear DNA analyses revealed a low level of genetic variability among spruce populations (only 3% of the total genetic variation (FST = 0.028) and a high variability within populations (97%). The total heterozygosity in all stands (HT) was calculated as 0.646. Based on UPGMA analysis, the most genetically similar populations are spruce stands in the Bieszczady National Park and the Ustrzyki Dolne Forest District, which have the smallest genetic divergence of all populations (DN = 0.0165). Our analysis of the mitochondrial gene nad1 revealed the presence of six different haplotypes “a”, “a1”, “b”, “c”, “d” and “d1”. Comprising 56% of all haplotypes, “a” was the most common showing a predominant impact on spruce migration from the Carpathian area. The analysis based on mitochondrial markers (by Nei) revealed a heterozygosity of 0.525. Based on the observations of disease symptoms, 29% of the trees belong to health class 1,30% to class 2,28% to class 3 and class 4 contains 13% of trees. The comparison between health status and the level of genetic variation in the analyzed stands showed a positive correlation. Spruce stands with better health were also characterized by a greater degree of genetic variability. Since most of the investigated spruce populations shared the mitochondrial haplotype “a”, we have ascertained their Hercynian- Carpathian origin. Only one stand (Cisna) had a high frequency (43.3%) of the Nordic haplotype “c” suggesting that this provenance is derived from the Baltic post-glacial refugium of P. abies in Europe.
This article provides morphological and molecular characteristics of Punctodera storiei Brzeski, 1998. Comparison of partial sequences of 18S and 28S rDNA genes from P. stonei sampled in Poland and Punctodera sp. from Canada showed their 100% similarity. This is the first report on the occurrence of P. stonei outside of Europe. We provide data on morphology of males and 2nd stage juveniles of this species and an identification key to males of the genus Punctodera Mulvey et Stone, 1976. Moreover, the paper presents evolutionary relationships of P. stonei within the family Heteroderidae.
Considered to be the most natural of all other Polish stands Picea abies in Białowieża Forest has suffered severely of bark beetle damages for decades. In order to find out some historical events in native Norway spruce stands diversity at regional level, we carried out the study on polymorphism with DNA markers and performed Principal Coordinate Analysis (PCoA) on the basis of genetic distance matrix. Two ranges (i.e. Baltico−Nordic and Hercyno−Carpathian) of Norway spruce cover north−eastern and southern Poland, respectively. The spruceless zone separates the these two ranges in lowland Poland. From genetic point of view, the Baltico−Nordic P. abies populations mainly harbor mitochondrial haplotype ‘c', whereas the spruces from the Hercyno−Carpathian range – haplotype ‘a'. Until recently, the Białowieża Forest has been considered to belong to the northern range of spruce occurrence in Europe. Seven populations from Białowieża Forest were studied (two of them from the Strict Reserve in Białowieski National Park) according to mitochondrial DNA diversity, and compared with genetic diversity found in 24 other populations located in Białystok Regional Directorate of the State Forests (north−eastern Poland). All studied spruce populations were more than 100 years old and were of natural origin. As result, the spruces from the Białowieża Forest harbored three mitochondrial haplotypes of the nad1 gene (‘c', ‘a' and ‘a1'), with the haplotype ‘a' being more frequently present (>51%) than the haplotype ‘c'. The spruce stands from Białowieża Forest have the highest gene diversity (h Nei=0.527) comparing to the other stands from the Białystok RDSF. The PCoA proved the particularity of the spruces from Białowieża Forest grouping them into one cluster of genetic similarity. Our data demonstrated that most of Norway spruces populations from Białystok RDSF harbor haplotype ‘c', which confirms their historical relationship with the Baltico−Nordic range of P. abies in Europe, while most spruces from Białowieża Forest have different historic origin, because they share high frequency of the southern haplotype ‘a'. It also turned out that Norway spruce populations from two different European ranges met in lowland Poland after the last glacial period, as proved by the presence of two mitochondrial haplotypes ‘a' and ‘c'. It can be assumed, that the spruceless zone was created by human activity in the past millennia. The results of all molecular analyses confirmed the unique character of spruces of Białowieża Forest, which is distinguished by the greater richness of the gene pool in comparison to the region of Białystok RDSF as well as good adaptation to local environmental conditions.
Since its description in 2001 Alcathoe's myotis (Myotis alcathoe) was recorded from several locations across Europe. Here we describe the first records of this species from Germany, Poland, Albania, and from the European part of Turkey, including the northernmost locality in central Germany (51°23′N, 11°01′E). Compilation of all up-to-date records shows that M. alcathoe has a wide European distribution although it seems to be rare at most places. The habitats where the bat was recorded are natural, moist and deciduous forests with old trees and water streams as can be found, for example, in canyons or forests of alluvial origin. Such habitats suggest that the species probably has a more continuous and wider distribution than currently known and might be expected to occur even further to the North.
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