Microsatellite Markers in Analysis of Forest‐Tree Populations

The present state of knowledge regarding the genetic diversity of forest tree species has been greatly improved with the development of the powerful research tool that the microsatellite markers represent. These noncoding sequences are considered to be neutral, highly polymorphic, and species specific. The usefulness of the microsatellite markers was recently proven by the determination of differentiation at inter‐ and intrapopulation level, gene flow in natural forest‐tree populations, heritability processes, and sustainable management of forest genetic resources in many natural forest stands. In this chapter, I aim to describe the practical approach of microsatellite markers, used in determination of genetic structure of 14 Scots pine populations from North‐eastern Poland. Investigated pine populations exhibited high genetic parameter variation, for example, mean PIC = 79.3, Shannon Index I = 2.488, observed (HO = 0.778) and expected (HE = 0.849) heterozygosity. Low level of Fst = 0.031 demonstrated that studied populations are more differentiated within than among stands, which were grouped into one cluster of genetic similarity. In conclusion, the present distribution of genetically related populations of Scots pine in North‐eastern Poland seems to reflect the historical events such as postglacial colonization of Poland from different European refugia and/or human management carried out in the past

Analysis of DNA profiles of ash (Fraxinus excelsior L.) to provide evidence of illegal logging

The present work formed part of a research project supported by the General Directorate of State Forests (Grants BLP-333 and BLP-384). We gratefully acknowledge the Forest Guard staff from Śnieżka Forest District for their efficient cooperation. We also thank Małgorzata Gorzkowska from the Laboratory of Molecular Biology FRI Poland, who assisted with processing of plant material in the laboratory.Peer reviewedPublisher PD

Postawa społeczeństwa wobec organizmów modyfikowanych genetycznie (GMO) w Polsce – na przykładzie wybranych grup osób

Poziom wiedzy społeczeństwa nt. GMO stale się zmienia. W listopadzie i grudniu 2019 r. studenci UKSW w Warszawie przeprowadzili ankiety wśród 699 anonimowych respondentów na temat zastosowania produktów GMO w codziennym życiu Polaków. Osoby ankietowane podzielono wg czterech kryteriów: płci, wieku, wykształcenia i miejsca zamieszkania z podziałem na miasto i wieś, zadając 7 pytań merytorycznych odnośnie do poziomu wiedzy i osobistego podejścia do konsumpcji produktów GMO. Analizę statystyczną wyników wykonano w programie STATISTICA 12. Wykazano, że w ostatnich latach rośnie wiedza mieszkańców w Polsce nt. GMO, ale obawy przed produktami genetycznie modyfikowanymi nadal występują u ponad połowy społeczeństwa. Większość kobiet obawia się GMO, zwraca większą uwagę na oznakowanie produktów zawierających GMO i nie podałaby ich swoim dzieciom. Osoby w wieku poniżej 20 lat są pozytywnie nastawione do GMO, natomiast osoby w wieku powyżej 35 lat obawiają się negatywnych skutków spożywania produktów GMO. Większość osób mieszkających na wsi obawia się produktów GMO, opinia zaś mieszkańców miasta jest podzielona na dwie, prawie równe grupy: zarówno zwolenników, jak i przeciwników GMO. Różnice w odpowiedziach odnośnie obawy przed negatywnymi skutkami spożywania produktów zawierających GMO były statystycznie istotne (p < 0,001) i wykazały większe obawy wśród osób zamieszkujących tereny wiejskie (67%) niż u mieszkańców miast (49%). Jednocześnie, prawie ten sam duży odsetek osób ze wsi i z miasta (67%) był przychylny stosowaniu leków wyprodukowanych drogą inżynierii genetycznej (p < 0,01)

Raman-based spectrophenotyping of the most important cells of the immune system

INTRODUCTION: Human peripheral blood mononuclear cells (PBMCs) are a heterogeneous population of cells that includes T and B lymphocytes. The total number of lymphocytes and their percentage in the blood can be a marker for the diagnosis of several human diseases. Currently, cytometric methods are widely used to distinguish subtypes of leukocytes and quantify their number. These techniques use cell immunophenotyping, which is limited by the number of fluorochrome-labeled antibodies that can be applied simultaneously. OBJECTIVE: B and T lymphocytes were isolated from peripheral blood obtained from healthy human donors. METHODS: The immunomagnetic negative selection was used for the enrichment of B and T cells fractions, and their purity was assessed by flow cytometry. Isolated cells were fixed with 0.5% glutaraldehyde and measured using confocal Raman imaging. K-means cluster analysis, principal component analysis and partial least squares discriminant methods were applied for the identification of spectroscopic markers to distinguish B and T cells. HPLC was the reference method for identifying carotene in T cells. RESULTS: Reliable discrimination between T and B lymphocytes based on their spectral profile has been demonstrated using label-free Raman imaging and chemometric analysis. The presence of carotene in T lymphocytes (in addition to the previously reported in plasma) was confirmed and for the first time unequivocally identified as β-carotene. In addition, the molecular features of the lymphocytes nuclei were found to support the discriminant analysis. It has been shown that although the presence of carotenoids in T cells depends on individual donor variability, the reliable differentiation between lymphocytes is possible based on Raman spectra collected from individual cells. CONCLUSIONS: This proves the potential of Raman spectroscopy in clinical diagnostics to automatically differentiate between cells that are an important component of our immune system

Global Geographic Distribution and Host Range of Fusarium circinatum, the Causal Agent of Pine Pitch Canker

Funding: This study was financially supported by COST Action FP1406 (PINESTRENGTH), the Estonian Science Foundation grant PSG136, the Forestry Commission, United Kingdom, the Phytophthora Research Centre Reg. No. CZ.02.1.01/0.0/0.0/15_003/0000453, a project co-financed by the European Regional Development Fund. ANSES is supported by a grant managed by the French National Research Agency (ANR) as part of the “Investissements d’Avenir” programme (ANR-11-LABX-0002-01, Laboratory of ExcellenceARBRE). SW was partly supported by BBSRC Grant reference BB/L012251/1 “Promoting resilience of UK tree species to novel pests & pathogens: ecological & evolutionary solutions (PROTREE)” jointly funded by BBSRC, Defra, ESRC, the Forestry Commission, NERC and the Scottish Government, under the Tree Health and Plant Biosecurity Initiative. Annual surveys in Switzerland were financially supported by the Swiss Federal Office for the Environment FOEN. Acknowledgments: Andrea Kunova and Cristina Pizzatti are acknowledged for the assistance in the sampling. Thanks are due to Dina Ribeiro and Helena Marques from ICNF-Portuguese Forest Authority for providing location coordinates. We thank three anonymous reviwers for valuable corrections and suggestions.Peer reviewedPublisher PD

Interactions between Phytophthora cactorum, Armillaria gallica and Betula pendula Roth. Seedlings Subjected to Defoliation

The purpose of this study was to better understand the interactive impact of two soil-borne pathogens, Phytophthora cactorum and Armillaria gallica, on seedlings of silver birch (Betula pendula Roth.) subjected to stress caused by mechanical defoliation, simulating primary insect feeding. This is the first experimental confirmation of silver birch seedling root damage (and in consequence shoot mortality) caused by the additive effect of defoliation stress and P. cactorum inoculation via soil. However, the most severe damage to roots occurred after A. gallica inoculation. One year after treatments, chlorophyll fluorescence measurement, and gas chromatography coupled with mass spectrometry (GC-MS) were used to analyze the photosynthetic activity in leaves, the volatile organic compounds (VOCs) emitted by the birch leaves, and chemical compounds from the roots. The cumulative effect of the two pathogens and partial defoliation reduced photosynthetic activity, suggesting dysfunction of photosystem PSII due to the applied stresses. In summary, it seems that the main differences in photosynthetic performance could be attributed to Armillaria infection. The birch leaves in seedlings exposed to 50% defoliation, and inoculation with P.cactorum and A. gallica, emitted more aromatic carbonyls and alcohols, as well as half as much aliphatic esters, compared to controls. In infected birch roots, the production of phenols, triterpenes, and fatty alcohols increased, but fatty acids decreased. Higher levels of aromatic carbonyls and alcohols in leaves, as well as phenolic compounds in the roots of stressed birches (compared to control) suggest an activation of plant systemic acquired resistance (SAR)

Antropogenic influence on the present genetic structure of forest tree populations

Human activity has always had an impact on the species composition and genetic structure of species that make up the forest ecosystem. The results of DNA-based studies confirm the influence of current and past forest management on the current level of genetic variability of forest tree populations. Molecular analysis enables the study of gene flow between generations and reflects the history of post-glacial migration of species from southern refugia of Europe to Poland. Thanks to DNA markers, it is possible to precisely distinguish species hybrids, e.g. between Japanese and European larches. In timber trade market, the comparison of the DNA profiles of a stolen tree with a stump left in the forest provides strong evidence in judiciary pursuits concerning the illegal logging. Genetic transformation of the tree genome allows to obtain wood with altered lignin structure, higher increment and increased assimilation of CO2 in the leaves. Genetic modifications are also aimed at enhanced immunity of cultured tree species to fungal pathogens or insect pests and to plant protection products on fast-growing GMO trees.Działalność człowieka od zawsze miała wpływ na skład gatunkowy i strukturę genetyczną gatunków tworzących ekosystem leśny. Wyniki badań DNA potwierdzają wpływ obecnej i przeszłej gospodarki leśnej na obecny poziom zmienności genetycznej populacji drzew leśnych. Analiza molekularna pozwala określić pokrewieństwo genetyczne gatunków drzew leśnych, umożliwia badanie przepływu genów między pokoleniami, oraz odzwierciedla historię polodowcowej migracji gatunków z refugiów południowej Europy do Polski. Dzięki markerom DNA można precyzyjnie odróżnić mieszańce gatunkowe np. modrzewia japońskiego i europejskiego. Porównanie profili DNA kradzionego drzewa z pniakiem pozostawionym w lesie, dostarcza mocnych dowodów w sprawach dotyczących nielegalnego handlu drewnem. Genetyczna transformacja genomu drzew pozwala uzyskać drewno o zmienionej strukturze lignin, oraz zwiększoną asymilację CO2 w liściach. Modyfikacje genetyczne zmierzają też w kierunku wzmocnionej odporności hodowanych gatunków drzew na patogeny grzybowe lub szkodniki owadzie oraz na środki ochrony roślin na plantacjach drzew szybkorosnących GMO

Determining invasiveness and risk of dothistroma needle blight

Determining invasiveness and risk of dothistroma needle bligh