Nina Elisabeth Nagy
Research Professor
Biography
Senior researcher at the Norwegian Institute of Bioeconomy Research (NIBIO, Div. Biotechnology and plant health). PhD in plant molecular biology (2002). Main research interests: plant-pathogen interactions, plant defense responses and bioprospecting. My research (with co-operation in Norway and abroad) has focused on anatomical and molecular resistance mechanisms in trees. This include how and where defense compounds act, which cell types and tissue regions are involved in their production as well as the molecular and genetic mechanisms behind. I am currently leading a project focusing on identifying chemical defense compounds in white rot spruce, with health application perspective. In addition to cell molecular biology, I have broad experience in microscopy techniques as light- and electron microscopy, laser micro dissection and imaging.
Authors
Fernanda Leiva Florent Abdelghafour Muath K Alsheikh Nina Elisabeth Nagy Jahn Davik Aakash ChawadeAbstract
Common scab (CS) is a major bacterial disease causing lesions on potato tubers, degrading their appearance and reducing their market value. To accurately grade scab-infected potato tubers, this study introduces “ScabyNet”, an image processing approach combining color-morphology analysis with deep learning techniques. ScabyNet estimates tuber quality traits and accurately detects and quantifies CS severity levels from color images. It is presented as a standalone application with a graphical user interface comprising two main modules. One module identifies and separates tubers on images and estimates quality-related morphological features. In addition, it enables the extraction of tubers as standard tiles for the deep-learning module. The deep-learning module detects and quantifies the scab infection into five severity classes related to the relative infected area. The analysis was performed on a dataset of 7154 images of individual tiles collected from field and glasshouse experiments. Combining the two modules yields essential parameters for quality and disease inspection. The first module simplifies imaging by replacing the region proposal step of instance segmentation networks. Furthermore, the approach is an operational tool for an affordable phenotyping system that selects scab-resistant genotypes while maintaining their market standards.
Abstract
Purpose of Review Forestry in northern temperate and boreal regions relies heavily on conifers. Rapid climate change and associated increases in adverse growing conditions predispose conifers to pathogens and pests. The much longer generation time and presumably, therefore, lower adaptive capacity of conifers relative to their native or non-native biotic stressors may have devastating consequences. We provide an updated overview of conifer defences underlying pathogen and pest resistance and discuss how defence traits can be used in tree breeding and forest management to improve resistance. Recent Findings Breeding of more resilient and stress-resistant trees will benefit from new genomic tools, such as genotyping arrays with increased genomic coverage, which will aid in genomic and relationship-based selection strategies. However, to successfully increase the resilience of conifer forests, improved genetic materials from breeding programs must be combined with more flexible and site-specific adaptive forest management. Summary Successful breeding programs to improve conifer resistance to pathogens and pests provide hope as well as valuable lessons: with a coordinated and sustained effort, increased resistance can be achieved. However, mechanisms underlying resistance against one stressor, even if involving many genes, may not provide any protection against other sympatric stressors. To maintain the adaptive capacity of conifer forests, it is important to keep high genetic diversity in the tree breeding programs. Choosing forest management options that include diversification of tree-species and forest structure and are coupled with the use of genetically improved plants and assisted migration is a proactive measure to increase forest resistance and resilience to foreseen and unanticipated biotic stressors in a changing climate.
Authors
Paal Krokene Isabella Børja Elena Carneros Toril Drabløs Eldhuset Nina Elisabeth Nagy Daniel Volařík Roman GebauerAbstract
Drought-induced mortality is a major direct effect of climate change on tree health, but drought can also affect trees indirectly by altering their susceptibility to pathogens. Here, we report how a combination of mild or severe drought and pathogen infection affected the growth, pathogen resistance and gene expression in potted 5-year-old Norway spruce trees [Picea abies (L.) Karst.]. After 5 weeks of drought, trees were inoculated with the fungal pathogen Endoconidiophora polonica. Combined drought–pathogen stress over the next 8 weeks led to significant reductions in the growth of drought-treated trees relative to well-watered trees and more so in trees subjected to severe drought. Belowground, growth of the smallest fine roots was most affected. Aboveground, shoot diameter change was most sensitive to the combined stress, followed by shoot length growth and twig biomass. Both drought-related and some resistance-related genes were upregulated in bark samples collected after 5 weeks of drought (but before pathogen infection), and gene expression levels scaled with the intensity of drought stress. Trees subjected to severe drought were much more susceptible to pathogen infection than well-watered trees or trees subjected to mild drought. Overall, our results show that mild drought stress may increase the tree resistance to pathogen infection by upregulating resistance-related genes. Severe drought stress, on the other hand, decreased tree resistance. Because drought episodes are expected to become more frequent with climate change, combined effects of drought and pathogen stress should be studied in more detail to understand how these stressors interactively influence tree susceptibility to pests and pathogens.
Authors
Chatchai Kosawang Isabella Børja Maria-Luz Herrero Nina Elisabeth Nagy Lene R. Nielsen Halvor Solheim Volkmar Timmermann Ari HietalaAbstract
Introduction: The ascomycete Hymenoscyphus fraxineus, originating from Asia, is currently threatening common ash (Fraxinus excelsior) in Europe, massive ascospore production from the saprotrophic phase being a key determinant of its invasiveness. Methods: To consider whether fungal diversity and succession in decomposing leaf litter are affected by this invader, we used ITS-1 metabarcoding to profile changes in fungal community composition during overwintering. The subjected ash leaf petioles, collected from a diseased forest and a healthy ash stand hosting the harmless ash endophyte Hymenoscyphus albidus, were incubated in the forest floor of the diseased stand between October 2017 and June 2018 and harvested at 2–3-month intervals. Results: Total fungal DNA level showed a 3-fold increase during overwintering as estimated by FungiQuant qPCR. Petioles from the healthy site showed pronounced changes during overwintering; ascomycetes of the class Dothideomycetes were predominant after leaf shed, but the basidiomycete genus Mycena (class Agaricomycetes) became predominant by April, whereas H. albidus showed low prevalence. Petioles from the diseased site showed little change during overwintering; H. fraxineus was predominant, while Mycena spp. showed increased read proportion by June. Discussion: The low species richness and evenness in petioles from the diseased site in comparison to petioles from the healthy site were obviously related to tremendous infection pressure of H. fraxineus in diseased forests. Changes in leaf litter quality, owing to accumulation of host defense phenolics in the pathogen challenged leaves, and strong saprophytic competence of H. fraxineus are other factors that probably influence fungal succession. For additional comparison, we examined fungal community structure in petioles collected in the healthy stand in August 2013 and showing H. albidus ascomata. This species was similarly predominant in these petioles as H. fraxineus was in petioles from the diseased site, suggesting that both fungi have similar suppressive effects on fungal richness in petiole/rachis segments they have secured for completion of their life cycle. However, the ability of H. fraxineus to secure the entire leaf nerve system in diseased forests, in opposite to H. albidus, impacts the general diversity and successional trajectory of fungi in decomposing ash petioles.
Abstract
After fungal decay experiments chemical characterisation of the wood is often a routine and several methodological approaches are available. In this study, we tested if simultaneous thermal analysis (STA) is a valid alternative to traditional wet chemical methods since STA allows significantly smaller sample size and faster analysis. Three model fungi including the brown rot fungi Rhodonia placenta and Gloeophyllum trabeum and the white rot fungus Trametes versicolor were employed in the study using Norway spruce as substrate. The experiment was harvested after 10, 20 and 52 weeks. At each harvest interval, aliquots of the material were characterized by STA and wet chemical methods. The results validated that STA can be effectively used to estimate cell wall composition of brown rot depolymerised wood. However, STA slightly overestimated cellulose at brown rot decay above 50%. The method was not verified for simultaneous white rot because STA only estimated hemicellulose correctly compared to the wet chemical method. Hence, STA is considered suitable for brown rot fungi below 50% mass loss but not for simultaneous white rot because STA did not estimate cellulose and lignin correctly.
Authors
Lene R. Nielsen Nina Elisabeth Nagy Sara Piqueras Chatchai Kosawang Lisbeth G. Thygesen Ari HietalaAbstract
Some common ash trees (Fraxinus excelsior) show tolerance towards shoot dieback caused by the invasive ascomycete Hymenoscyphus fraxineus. Leaf petioles are considered to serve as a pathogen colonization route to the shoots. We compared four common ash clones with variation in disease tolerance, and included the native host, Manchurian ash (Fraxinus mandshurica), as a reference. Tissue colonization, following rachis inoculation by H. fraxineus, was monitored by histochemical observations and a quantitative polymerase chain reaction (qPCR) assay specific to H. fraxineus. Axial spread of the pathogen towards the petiole base occurred primarily within the phloem and parenchyma, tissues rich in starch in healthy petioles. In inoculated petioles, a high content of phenolics surrounded the hyphae, presumably a host defense response. There was a relationship between field performance and susceptibility to leaf infection in three of the four studied common ash clones, i.e., good field performance was associated with a low petiole colonization level and vice versa. Low susceptibility to leaf infection may counteract leaf-to-shoot spread of the pathogen in common ash, but the limited number of clones studied warrants caution and a larger study. The Manchurian ash clone had the highest petiole colonization level, which may suggest that this native host has evolved additional mechanisms to avoid shoot infection.
Authors
Nina Elisabeth Nagy Hans Ragnar Norli Monica Fongen Runa Berg Østby Inger Heldal Jahn Davik Ari HietalaAbstract
Tree defense against xylem pathogens involves both constitutive and induced phenylpropanoids and terpenoids. The induced defenses include compartmentalization of compromised wood with a reaction zone (RZ) characterized by polyphenol deposition, whereas the role of terpenoids has remained poorly understood. To further elucidate the tree–pathogen interaction, we profiled spatial patterns in lignan (low-molecular-weight polyphenols) and terpenoid content in Norway spruce (Picea abies) trees showing heartwood colonization by the pathogenic white-rot fungus Heterobasidion parviporum. There was pronounced variation in the amount and composition of lignans between different xylem tissue zones of diseased and healthy trees. Intact RZ at basal stem regions, where colonization is the oldest, showed the highest level and diversity of these compounds. The antioxidant properties of lignans obviously hinder oxidative degradation of wood: RZ with lignans removed by extraction showed significantly higher mass loss than unextracted RZ when subjected to Fenton degradation. The reduced diversity and amount of lignans in pathogen-compromised RZ and decaying heartwood in comparison to intact RZ and healthy heartwood suggest that α-conindendrin isomer is an intermediate metabolite in lignan decomposition by H. parviporum. Diterpenes and diterpene alcohols constituted above 90% of the terpenes detected in sapwood of healthy and diseased trees. A significant finding was that traumatic resin canals, predominated by monoterpenes, were commonly associated with RZ. The findings clarify the roles and fate of lignan during wood decay and raise questions about the potential roles of terpenoids in signal transduction, synthesis, and translocation of defense compounds upon wood compartmentalization against decay fungi.
Authors
Ari Hietala Ahto Agan Nina Elisabeth Nagy Isabella Børja Volkmar Timmermann Rein Drenkhan Halvor SolheimAbstract
The populations of European ash and its harmless fungal associate Hymenoscyphus albidus are in decline owing to ash dieback caused by the invasive Hymenoscyphus fraxineus, a fungus that in its native range in Asia is a harmless leaf endophyte of local ash species. To clarify the behavior of H. albidus and its spatial and temporal niche overlap with the invasive relative, we used light microscopy, fungal species-specific qPCR assays, and PacBio long-read amplicon sequencing of the ITS1-5.8S-ITS2 region to examine fungal growth and species composition in attached leaves of European ash. The plant material was collected from a healthy stand in central Norway, where ash saplings in late autumn showed leaflet vein necrosis like that commonly related to H. fraxineus. For reference, leaflet samples were analyzed from stands with epidemic level of ash dieback in southeastern Norway and Estonia. While H. albidus was predominant in the necrotic veins in the healthy stand, H. fraxineus was predominant in the diseased stands. Otherwise, endophytes with pathogenic potential in the genera Venturia (anamorph Fusicladium), Mycosphaerella (anamorph Ramularia), and Phoma, and basidiomycetous yeasts formed the core leaflet mycobiome both in the healthy and diseased stands. In necrotic leaf areas with high levels of either H. albidus or H. fraxineus DNA, one common feature was the high colonization of sclerenchyma and phloem, a region from which the ascomata of both species arise. Our data suggest that H. albidus can induce necrosis in ash leaves, but that owing to low infection pressure, this first takes place in tissues weakened by autumn senescence, 1–2 months later in the season than what is characteristic of H. fraxineus at an epidemic phase of ash dieback. The most striking difference between these fungi would appear to be the high fecundity of H. fraxineus. The adaptation to a host that is phylogenetically closely related to European ash, a tree species with high occurrence frequency in Europe, and the presence of environmental conditions favorable to H. fraxineus life cycle completion in most years may enable the build-up of high infection pressure and challenge of leaf defense prior to autumn senescence.
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This study explores cell wall changes in Radiata pine (Pinus radiata) after modification with acetylation or furfurylation and subsequent prolonged subjection to the brown rot fungus R. placenta with the aim of better understanding the modus operandi of these two modifications. Both modifications have shown good durability in field tests, but in order to learn from their possible limitations, we used optimal environmental conditions for fungal growth, and extended the testing period compared to standard tests. Hyphae were found in acetylated wood after two weeks, and after 28 weeks of decay abundant amounts of encapsulated hyphae were present. In furfurylated wood, mass loss and a few hyphae were seen initially, but no further development was seen during weeks 18–42. The general degradation pattern was qualitatively the same for unmodified, acetylated and furfurylated wood: carbohydrates decreased relative to lignin. Acetyl groups were lost from acetylated wood during decay (earlier results), while the furan polymer did not seem to be altered by the fungus. Based on these findings it is hypothesized that modifications such as furfurylation that enhance moisture exclusion within the cell wall through impregnation polymerization offer better long term protection compared to modifications such as acetylation that depend on the replacement of hydroxyl groups with ether bound adducts that can be removed by fungi.
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The scope of this study was to provide an update on fluoride (F) emission effects on vegetation around three aluminium smelters. We visited Årdal and Sunndal smelters in 2019-2020 and Mosjøen in 2020, assessed and documented the visual symptoms of F-damage on vegetation and related these to detected values of F in plant tissue. Three plant species showed qualities as useful indicators: Rowan, pine and St. John’s wort. Because male-fern accumulated extreme F-values and showed clear grazing damage, the monitoring of this species may be warranted because of the potential health hazard for the grazing animals. In Årdal and Sunndal, during 2019 and 2020, we detected the highest F-values in male-fern, ranging from 94 to 925 mg F/kg. In rowan, the highest F-concentration was detected in trees growing within the Årdal smelter (1161 mg F/kg) but on all other locations the F-concentrations in rowan ranged from 4 to 327 mg F/kg. In pine, the F-concentrations ranged from 6-351 mg F/kg for all needle ages, but older needles always accumulated more F than younger ones. In St. John’s wort the accumulated F-values ranged from 10-84 mg F/kg. At all smelters there was a gradient of decreasing F-concentration in vegetation with increasing distance from the smelter. F-emissions in Årdal (12 and 11 kg F/hour in 2019 and 2020, respectively) and in Sunndal during 2019 (12 kg F/hour) were only slightly higher than the recommended limits (10 kg F/hour) for damage on vegetation, while in Mosjøen the F-emissions were 7 kg F/hour in 2020. The presence of F-damage on vegetation was consistent with the reported emission-levels. On basis of this evaluation, reductions in emissions are still advisable in Årdal and Sunndal, while the situation is acceptable in Mosjøen.
Authors
Ahto Agan Rein Drenkhan Kalev Adamson Leho Tedersoo Halvor Solheim Isabella Børja Iryna Matsiakh Volkmar Timmermann Nina Elisabeth Nagy Ari HietalaAbstract
European ash (Fraxinus excelsior) is threatened by the invasive ascomycete Hymenoscyphus fraxineus originating from Asia. Ash leaf tissues serve as a route for shoot infection but also as a sporulation substrate for this pathogen. Knowledge of the leaf niche partitioning by indigenous fungi and H. fraxineus is needed to understand the fungal community receptiveness to the invasion. We subjected DNA extracted from unwashed and washed leaflets of healthy and diseased European ash to PacBio sequencing of the fungal ITS1-5.8S-ITS2 rDNA region. Leaflets from co-inhabiting rowan trees (Sorbus aucuparia) served as a reference. The overlap in leaflet mycobiomes between ash and rowan was remarkably high, but unlike in rowan, in ash leaflets the sequence read proportion, and the qPCR-based DNA amount estimates of H. fraxineus increased vigorously towards autumn, concomitant with a significant decline in overall fungal richness. The niche of ash and rowan leaves was dominated by epiphytic propagules (Vishniacozyma yeasts, the dimorphic fungus Aureobasidion pullulans and the dematiaceous hyphomycete Cladosporium ramotenellum and H. fraxineus), and endophytic thalli of biotrophs (Phyllactinia and Taphrina species), the indigenous necrotroph Venturia fraxini and H. fraxineus. Mycobiome comparison between healthy and symptomatic European ash leaflets revealed no significant differences in relative abundance of H. fraxineus, but A. pullulans was more prevalent in symptomatic trees. The impacts of host specificity, spatiotemporal niche partitioning, species carbon utilization profiles and life cycle traits are discussed to understand the ecological success of H. fraxineus in Europe. Further, the inherent limitations of different experimental approaches in the profiling of foliicolous fungi are addressed.
Authors
Marit Almvik Nina Elisabeth Nagy Hans Ragnar Norli Ari Hietala Sven R. Odenmarck Monica Fongen Anas KamlehAbstract
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Authors
Roman Gebauer Daniel Volařík Josef Urban Isabella Børja Nina Elisabeth Nagy Toril Drabløs Eldhuset Paal KrokeneAbstract
Several studies have looked at how individual environmental factors influence needle morphology in conifer trees, but interacting effects between drought and canopy position have received little attention. In this study, we characterized morphological responses to experimentally induced drought stress in sun exposed and shaded current-year Norway spruce needles. In the drought plot trees were suffering mild drought stress, with an average soil water potential at 50 cm depth of -0.4 MPa. In general, morphological needle traits had greater values in sun needles in the upper canopy than in shaded needles in the lower canopy. Needle morphology 15 months after the onset of drought was determined by canopy position, as only sun needle morphology was affected by drought. Thus, canopy position was a stronger morphogenic factor determining needle structure than was water availability. The largest influence of mild drought was observed for needle length, projected needle area and total needle area, which all were reduced by ~27% relative to control trees. Needle thickness and needle width showed contrasting sensitivity to drought, as drought only affected needle thickness (10% reduction). Needle dry mass, leaf mass per area and needle density were not affected 15 months after the onset of mild drought. Our results highlight the importance of considering canopy position as well as water availability when comparing needle structure or function between conifer species. More knowledge about how different canopy parts of Norway spruce adapt to drought is important to understand forest productivity under changing environmental conditions.
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Authors
Marit Almvik Marie-Pierre Pavageau Nina Elisabeth Nagy Hans Ragnar Norli Ari Hietala Sven R. Odenmarck Monica Fongen Anas KamlehAbstract
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Ari Hietala Isabella Børja Hugh B. Cross Nina Elisabeth Nagy Jørn Henrik Sønstebø Volkmar Timmermann Adam Vivian-Smith Halvor SolheimAbstract
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Authors
Ari Hietala Isabella Børja Hugh Cross Nina Elisabeth Nagy Halvor Solheim Volkmar Timmermann Adam Vivian-SmithAbstract
European ash (Fraxinus excelsior), a keystone species with wide distribution and habitat range in Europe, is threatened at a continental scale by an invasive alien ascomycete, Hymenoscyphus fraxineus. In its native range of Asia, this fungus is a leaf endophyte with weak parasitic capacity and robust saprobic competence in local ash species that are closely related to European ash. In European ash, H. fraxineus has a similar functional role as in Asia, but the fungus also aggressively kills shoots, resulting in crown dieback and tree death. H. fraxineus is a typical invasive species, as its spread relies on high propagule pressure. While crown dieback of European ash is the most obvious symptom of ash dieback, the annual colonization of ash leaves is a crucial key dependency for the invasiveness of H. fraxineus, since its fruiting bodies are formed on overwintered leaf vein tissues in soil debris. Leaves of European ash host a wide range of indigenous epiphytes, endophytes, facultative parasites and biotrophic fungi, including Hymenoscyphus albidus, a relative of H. fraxineus that competes for the same sporulation niche as the invader. At face value, leaves of European ash are colonized by a large and diverse indigenous mycobiome. In order to understand why this invader became successful in Europe, we discuss and summarize the current knowledge of diversity, seasonal dynamics and traits of H. fraxineus and indigenous fungi associated with leaves of European ash.
Authors
Nina Elisabeth NagyAbstract
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Dieback of European ash, caused by the ascomycete Hymenoscyphus fraxineus originating from Asia, has rapidly spread across Europe, and is threatening this keystone tree at a continental scale. High propagule pressure is characteristic to invasive species. Consistently, the enormous production of windborne ascospores by H. fraxineus in an ash forest with epidemic level of disease obviously facilitates its invasiveness and long distance spread. To understand the rate of build-up of propagule pressure by this pathogen following its local introduction, during 2011–2017 we monitored its sporulation at a newly infested ash stand in south-western Norway characterized with mild winters and cool summers. We also monitored the propagule pressure by Hymenoscyphus albidus, a non-pathogenic native species that competes for the same sporulation niche with H. fraxineus. During the monitoring period, crown condition of ash trees had impaired, and 20% of the dominant trees were severely damaged in 2017. H. fraxineus showed an exponential increase in spore production between 2012 and 2015, followed by drastic decline in 2016 and 2017. During 2011–2013, the two Hymenoscyphus species showed similar sporulation level, but thereafter spores of H. albidus were no longer detected. The data suggest that following local introduction, the population of H. fraxineus reaches rapidly an exponential growth stage if the local weather conditions are favorable for ascomata maturation across years. In the North Atlantic climate, summer temperatures critically influence the pathogen infection pressure, warm summers allowing the population to grow according to its biotic potential, whereas cold summers can cause a drastic decline in propagule pressure.
Authors
Isabella Børja Volkmar Timmermann Ari Hietala Mari Mette Tollefsrud Nina Elisabeth Nagy Adam Vivian-Smith Hugh Cross Jørn Henrik Sønstebø Tor Myking Halvor SolheimAbstract
In Norway the common ash (Fraxinus excelsior L.) has its northernmost distribution in Europe. It grows along the coastal range as small fragmented populations. The first occurrence of ash dieback caused by Hymenoscyphus fraxineus in Norway was reported in 2008. At that time, the disease had already spread through large areas of southern and south-eastern parts of Norway. Since then the disease continued spreading with a speed of about 50- 60 km per year along the western coastal range. To monitor the disease development over time, we established eight permanent monitoring plots in south-eastern and western Norway in 2009 and 2012, respectively. In all plots tree mortality was high, especially among the youngest trees in south-eastern Norway. The extent of crown damage has continually increased in all diameter classes for both regions. In 2009, 76.8 % of all trees on the five monitoring plots in south-eastern Norway were considered to be healthy or slightly damaged, and only 8.9 % to be severely damaged. In 2015, 51.7 % were dead, 13.5 % severely damaged and only 25.7 % remained healthy or slightly damaged. To assess the infection pressure and spore dispersal patterns of the pathogen, we used a Burkard volumetric spore sampler placed in an infested ash stand in southern Norway. We examined the airborne ascospores of H. fraxineus and H. albidus captured on the sampling tape microscopically and with real-time PCR assays specific to these fungi. We detected very few ascospores of H. albidus, whereas ascospores of H. fraxineus dominated throughout entire sampling periods of 2009, 2010 and 2011. Spore discharge occurred mainly between the hours of 5 and 8 a.m., though the distinctive sporulation had yearly variation between 5-7 a.m. We observed the same diurnal pattern throughout the entire sampling period, with a seasonal peak in spore liberation between mid-July and midAugust, after which the number of ascospores decreased substantially. Similar diurnal patterns were observed throughout the sampling period except that after mid-August the number of trapped ascospores substantially decreased. To compare the genetic pattern of common ash in the northern and central ranges of Europe we analyzed the Norwegian samples together with available samples from central Europe by using chloroplast and nuclear microsatellite markers. We found that the northern range of common ash was colonized via a single migration route that originated in eastern or south-eastern Europe with little influence originating from other southern or western European refugia. In the northern range margins, genetic diversity decreased and population differentiation increased, coherent with a post-glacial colonization history characterized by founder events and population fluctuations. Based on our findings we discuss the future management and conservational implications.
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Dieback of European ash (Fraxinus excelsior L.), a disease caused by the ascomycete Hymenoscyphus fraxineus (previously referred to as H. pseudoalbidus or Chalara fraxinea), was first observed in Poland in the early 1990ies, and is currently present almost throughout the entire distribution area of European ash. The characteristic symptoms of the disease include dead shoots with necrotic lesions in the bark and discoloration of xylem and pith but the seasonal dynamics of pathogen spread in shoot tissues remain poorly understood. To investigate whether the internal spread of the fungus involves season-specific patterns, saplings with necrotic bark lesions in 1-2 -year-old stem regions were collected during 2014-2015 at time intervals in spring, summer, autumn and winter at several localities in western Ukraine and at two localities in south-eastern Norway. Tissuespecific presence of H. fraxineus was determined by a highly sensitive quantitative real-time PCR assay that is specific to DNA of H. fraxineus. The relatively high proportion of bark samples positive for H. fraxineus in the saplings collected during spring provides support to a model that H. fraxineus can be a primary causative agent of bark lesions and that other fungi may eventually replace it in old infection areas.
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Authors
Isabella Børja Douglas L. Godbold Jan Světlík Nina Elisabeth Nagy Roman Gebauer Josef Urban Daniel Volařík Holger Lange Paal Krokene Petr Čermák Toril Drabløs EldhusetAbstract
Global warming will most likely lead to increased drought stress in forest trees. We wanted to describe the adaptive responses of fine roots and fungal hyphae, at different soil depths, in a Norway spruce stand to long-term drought stress induced by precipitation exclusion over two growing seasons. We used soil cores, minirhizotrons and nylon meshes to estimate growth, biomass and distribution of fine roots and fungal hyphae at different soil depths. In control plots fine roots proliferated in upper soil layers, whereas in drought plots there was no fine root growth in upper soil layers and roots mostly occupied deeper soil layers. Fungal hyphae followed the same pattern as fine roots, with the highest biomass in deeper soil layers in drought plots. We conclude that both fine roots and fungal hyphae respond to long-term drought stress by growing into deeper soil layers.
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Ash dieback, caused by the ascomycete Hymenoscyphus fraxineus, has been spreading throughout Europe since the early 1990s, threatening European ash at a continental scale. Little is known about the development of the disease in individual forest trees and in different age classes. In this study we monitored ash dieback on trees of different diameter classes in five permanent plots in ash stands in south-eastern Norway from 2009 to 2016, and from 2012 to 2016 in three plots in western Norway with a shorter disease history. Our results showed that more than 80% of the youngest and more than 40% of the intermediate future crop trees in the plots in south-eastern Norway were dead by 2016, while the disease development in large, dominant trees was slower. Although less damage has been observed in the plots in western Norway, the trend for the juvenile trees is the same as in south-eastern Norway with rapidly increasing damage and mortality. Most dead trees in south-eastern Norway were found at sites with high soil moisture and showed symptoms of root-rot caused by Armillaria species. Infected trees, both young and old ones, are weakened by the disease and appear to be more susceptible to other, secondary pathogens, especially under unfavourable site conditions.
Authors
Hugo de Boer Maria Gulbrandsen Asmyhr Hanne Hegre Grundt Kjersti Sjøtun Hans K. Stenøien Jan Ove Gjershaug Kjetil Hindar Lawrence R. Kirkendall Nina Elisabeth Nagy Anders Nielsen Eli Knispel Rueness Odd Terje Sandlund Gaute VelleAbstract
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Anders Nielsen Merethe Aasmo Finne Maria Gulbrandsen Asmyhr Jan Ove Gjershaug Lawrence R. Kirkendall Vigdis Vandvik Gaute Velle Hugo de Boer Kjetil Hindar Nina Elisabeth Nagy Eli Knispel Rueness Odd Terje Sandlund Kjersti Sjøtun Hans K. StenøienAbstract
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Elena Carneros Nina Elisabeth Nagy Marcos Viejo Igor A. Yakovlev Jorunn Elisabeth Olsen Carl Gunnar FossdalAbstract
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Ari Hietala Hugh Cross Halvor Solheim Volkmar Timmermann Nina Elisabeth Nagy Isabella Børja Adam Vivian-Smith Jørn Henrik SønstebøAbstract
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Hugh Cross Jørn Henrik Sønstebø Nina Elisabeth Nagy Volkmar Timmermann Halvor Solheim Isabella Børja Håvard Kauserud Tor Carlsen Barbara Rzepka Katarzyna Wasak Adam Vivian-Smith Ari HietalaAbstract
High biodiversity is regarded as a barrier against biological invasions. We hypothesized that the invasion success of the pathogenic ascomycete Hymenoscyphus fraxineus threatening common ash in Europe relates to differences in dispersal and colonization success between the invader and the diverse native competitors. Ash leaf mycobiome was monitored by high-throughput sequencing of the fungal internal transcribed spacer region (ITS) and quantitative PCR profiling of H. fraxineus DNA. Initiation of ascospore production by H. fraxineus after overwintering was followed by pathogen accumulation in asymptomatic leaves. The induction of necrotic leaf lesions coincided with escalation of H. fraxineus DNA levels and changes in proportion of biotrophs, followed by an increase of ubiquitous endophytes with pathogenic potential. H. fraxineus uses high propagule pressure to establish in leaves as quiescent thalli that switch to pathogenic mode once these thalli reach a certain threshold – the massive feedback from the saprophytic phase enables this fungus to challenge host defenses and the resident competitors in mid-season when their density in host tissues is still low. Despite the general correspondence between the ITS-1 and ITS-2 datasets, marker biases were observed, which suggests that multiple barcodes provide better overall representation of mycobiomes.
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Modified wood can provide protection against a range of wood deteriorating organisms. Several hypotheses have been put forward regarding the protection mechanisms against wood decaying fungi including fungal enzyme inefficiency due to non-recognition, lower micropore size, and insufficient wood moisture content. The aim of this study was to obtain new insight into the protection manner of furfuryl alcohol (FA) modified Scots pine sapwood (WFA), and to examine biochemical mechanisms and adaptive changes in gene expression utilised by Postia placenta during early colonisation of WFA. Samples were harvested after 2, 4, and 8 weeks of incubation. After 8 weeks, the mass loss (0.1%) and wood moisture content (21.0%) was lower inWFA, than in non-modified Scots pine sapwood samples (W), 26.1% and 46.1%, respectively. Microscopy revealed needle-shaped calcium oxalate crystals, at all harvesting points, most prominently present after 4 and 8 weeks, and only in the WFA samples. Among the findings based on gene profiles were indications of a possible shift toward increased expression, or at least no down regulation, of genes related to oxidative metabolism and concomitant reduction of several genes related to the breakdown of polysaccharides in WFA compared to W.
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Authors
Roman Gebauer Daniel Volařík Josef Urban Isabella Børja Nina Elisabeth Nagy Toril Drabløs Eldhuset Paal KrokeneAbstract
No abstract has been registered
Authors
Ari Hietala Isabella Børja Hugh Cross Volkmar Timmermann Nina Elisabeth Nagy Halvor Solheim Jørn Henrik Sønstebø adam vivian smithAbstract
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Authors
Ari Hietala Isabella Børja Hugh Cross Nina Elisabeth Nagy Halvor Solheim Jørn Henrik Sønstebø Volkmar Timmermann Adam Vivian-SmithAbstract
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Abstract
Pythium species are fungal-like organisms distributed all over the world. Most Pythium spp. live as saprophytes, but some of them are pathogenic. Here we report on disease incidence in Norway spruce (Picea abies) seedlings caused by Pythium undulatum, and pathogenicity in vitro of Norwegian isolates of P. undulatum and P. anandrum.
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Authors
Ari Hietala Hugh Cross Isabella Børja Nina Elisabeth Nagy Volkmar Timmermann Halvor Solheim Jørn Henrik Sønstebø Adam Vivian-SmithAbstract
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Abstract
Ten saplings of European ash (Fraxinus excelsior L.) naturally infected by the invasive ash dieback pathogen Hymenoscyphus fraxineus were collected in Ukraine and Norway and examined for bark necrosis and extension of discoloration in sapwood and pith in a stem region. Tissue-specific colonization profiles were determined by spatial analyses of symptomatic and visually healthy stem tissues using a H. fraxineus-specific qPCR assay and light microscopy. Our data suggest that hyphal growth in the starch-rich perimedullary pith is of particular importance for both axial and radial spread of H. fraxineus, but that most of its biomass accumulates in sapwood parenchyma. The study confirms the results from earlier work and presents new information that refines the current stem invasion model.
Authors
Toril Drabløs Eldhuset Nina Elisabeth Nagy Isabella Børja Igor A. Yakovlev Roman Gebauer Daniel Volarik Paal KrokeneAbstract
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Authors
Nina Elisabeth Nagy Katarzyna Sikora Paal Krokene Ari Hietala Halvor Solheim Carl Gunnar FossdalAbstract
No abstract has been registered
Authors
Roman Gebauer Daniel Volařík Josef Urban Isabella Børja Nina Elisabeth Nagy Toril Drabløs Eldhuset Paal KrokeneAbstract
Trees must respond to many environmental factors during their development, and light is one of the main stimuli regulating tree growth. Thinning of forest stands by selective tree removal is a common tool in forest management that increases light intensity. However, morphological and anatomical adaptations of individual shoots to the new environmental conditions created by thinning are still poorly understood. In this study, we evaluated shoot morphology (shoot length, needle number, projected leaf area) and anatomy (tracheid lumen area, tracheid number, tracheid dimensions, xylem area, potential hydraulic conductivity) in three Norway spruce (Picea abies/L./Karst.) families exposed to different thinning regimes. We compared shoot characteristics of upper-canopy (i.e. sun-exposed) and lower-canopy (i.e. shaded) current-year shoots in a control plot and a plot thinned to 50 % stand density the previous year. One tree per family was chosen in each treatment, and five shoots were taken per canopy position. We found that upper-canopy shoots in both plots had higher values than lower-canopy shoots for all studied parameters, except lumen roundness and tracheid frequency (i.e. tracheid number per xylem area). Thinning had little effect on shoot morphology and anatomy 1 year after thinning, except for small but significant changes in tracheid dimensions. Needles were more sensitive to altered light conditions, as projected leaf area of shoot, needle number and leaf hydraulic conductivity changed after thinning. Differences between upper- and lower-canopy shoots did not seem to be influenced by thinning and were almost the same in both plots. Our results suggest that lower-canopy shoots require several years to modify their morphology and anatomy to new light conditions following thinning. The slow light adaptation of the lower canopy may be of practical importance in forest management: thinned stands may be predisposed to drought stress because newly exposed shoots experience increased illumination and transpiration after thinning.
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Authors
Isabella Børja Douglas Lawrence Godbold Michael Bambrick Lise Dalsgaard Jan Svetlik Nina Elisabeth NagyAbstract
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Abstract
Local climate conditions have a major influence on the biological decomposition of wood. To examine the influence of different temperature regimes on wood decay caused by the brown rot fungus Postia placenta in wood with differing natural durability, sapwood (sW) and heartwood (hW) of Scots pine, inoculated mini-blocks were incubated for up to 10 weeks at temperatures conducive or above optimal to wood decay. We profiled mass loss (ML) and wood composition, and accompanying changes in wood colonization and transcript level regulation of fungal candidate genes. The suppressive effect of suboptimal temperature on wood decay caused by P. placenta appeared more pronounced in Scots pine hW with increased durability than in sW with low decay resistance. The differences between sW and hW were particularly pronounced for cultures incubated at 30°C: unlike sW, hW showed no ML, poor substrate colonization and marker gene transcript level profiles indicating a starvation situation. As brown rot fungi show considerable species-specific variation in temperature optima and ability to mineralize components that contribute to wood durability, interactions between these factors will continue to shape the fungal communities associated to wood in service.
Abstract
This chapter provides an overview of anatomical and ecological aspects of resin-based defences in pines and contrasts the defence strategy of pines with that of other conifers. The main constituents of conifer resin are mono- and diterpenes in about equal amounts, with smaller amounts of sesquiterpenes. Resin production and storage represent a great cost for the trees, and because resin is both chemically toxic and physically deterring to insects and pathogens it has long been considered an important defence mechanism in conifers. Preformed or constitutive resin structures are present in pines and all other members of the pine family, but are generally absent in non-Pinaceae species. Resin stored under pressure in constitutive ducts flows out when a tree is injured and helps trapping or repelling invading organisms and sealing the wound. Pines have constitutive resin ducts in needles, phloem and xylem. In the phloem and xylem constitutive resin ducts are oriented both radially (within the radial rays) and axially in the form of cortical resin ducts in the outer phloem and constitutive resin ducts in the xylem. Numerous connections between the radial resin ducts and the axial resin ducts in the xylem create a large inter-connected resin reservoir. In addition, so-called traumatic resin ducts can be induced axially in the xylem in response to wounding, insect attack or other biotic and abiotic stresses. Traumatic resin ducts may contribute to so-called acquired or systemic induced resistance that increases tree resistance to future attacks.
Authors
Toril Drabløs Eldhuset Nina Elisabeth Nagy Daniel Volařík Isabella Børja Roman Gebauer Igor A. Yakovlev Paal KrokeneAbstract
Purpose: Drought-induced tree susceptibility is a major risk associated with climate change. Here we report how an 11-week drought affected tracheid structure, gene expression, and above- and belowground growth in 5-year-old Norway spruce trees (Picea abies) under controlled conditions. Results: The canopy of trees subjected to severe drought had significantly less current-year needle biomass, and fewer tracheids and tracheid rows in current-year shoots compared to fully watered control trees. Belowground tissues were more strongly affected by drought than aboveground tissues. In fine roots (<2 mm diameter) severe drought significantly reduced root biomass, root diameter, root length density and root surface area per soil volume compared to the control. Tracheid diameter and hydraulic conductivity in fine roots were significantly lower and tracheid flatness higher in trees subjected to severe drought than in control trees, both for long and short roots. Transcripts of the drought-related dehydrins PaDhn1 and PaDhn6 were strongly upregulated in stem bark and current-year needles in response to drought, whereas PaDhn4.5 was down-regulated. Conclusions: This study demonstrates that drought reduces biomass and hydraulic conductivity in fine roots and needles. We suggest that the ratio between PaDhn6 and PaDhn4.5 may be a sensitive marker of drought stress in Norway spruce.
Authors
Toril Drabløs Eldhuset Isabella Børja Roman Gebauer Paal Krokene Nina Elisabeth Nagy Josef Urban Daniel VolarikAbstract
No abstract has been registered
Authors
Roman Gebauer Daniel Volařík Josef Urban Isabella Børja Nina Elisabeth Nagy Toril Eldhuset Paal KrokeneAbstract
Conifer needles are extraordinarily variable and much of this diversity is linked to the water transport capacity of the xylem and to xylem conduit properties. However, we still know little about how anatomical characteristics influence the hydraulic efficiency of needle xylem in different parts of the crown. In this study we evaluated needle function and anatomy in Norway spruce families exposed to different light conditions. We measured tracheid and needle characteristics of sun-exposed and shaded current-year needles in two experimental plots: a control plot and a thinned plot with 50% reduction in stand density. Sun-exposed needles had a larger tracheid lumen area than shaded needles, and this was caused by a larger maximum tracheid lumen diameter, while the minimum lumen diameter was less plastic. Sun-exposed needles had also higher theoretical hydraulic conductivity than shaded needles. Thinning leads to increased radiation to the lower branches, and presumably exposes the upper branches to stronger water stress than before thinning. Thinning affected several needle parameters both in sun-exposed and shaded needles; tracheid lumens were more circular and minimum tracheid lumen diameter was larger in the thinned plot, whereas maximum tracheid lumen diameter was less plastic on both plots. This study demonstrates that needle xylem structure in Norway spruce is clearly influenced by the light gradient within the tree crown.
Authors
Simon Ballance Stefan Sahlstrøm Per Lea Nina Elisabeth Nagy Petter V. Andersen Tzvetelin Dessev Sarah Hull Maria Vardakou Richard FaulksAbstract
Purpose: To identify the key parameters involved in cereal starch digestion and associated glycaemic response by the utilisation of a dynamic gastro-duodenal digestion model. Methods: Potential plasma glucose loading curves for each meal were calculated and fitted to an exponential function. The area under the curve (AUC) from 0 to 120 min and total digestible starch was used to calculate an in vitro glycaemic index (GI) value normalised against white bread. Microscopy was additionally used to examine cereal samples collected in vitro at different stages of gastric and duodenal digestion. Results: Where in vivo GI data were available (4 out of 6 cereal meals) no significant difference was observed between these values and the corresponding calculated in vitro GI value. Conclusion: It is possible to simulate an in vivo glycaemic response for cereals when the gastric emptying rate (duodenal loading) and kinetics of digestible starch hydrolysis in the duodenum are known.
Abstract
The outcome of a compatible mycorrhizal interaction is different from that in a compatible plant–pathogen interaction; however, it is not clear what mechanisms are used to evade or suppress the host defence. The aim of this work is to reveal differences between the interaction of Norway spruce roots to the pathogen Ceratocystis polonica and the ectomycorrhizal Laccaria bicolor, examine if L. bicolor is able to evade inducing host defence responses typically induced by pathogens, and test if prior inoculation with the ectomycorrhizal fungus affects the outcome of a later challenge with the pathogen. The pathogen was able to invade the roots and caused extensive necrosis, leading to seedling death, with or without prior inoculation with L. bicolor. The ectomycorrhizal L. bicolor colonised primary roots of the Norway spruce seedlings by partly covering, displacing and convoluting the cells of the outer root cortex, leaving the seedlings healthy. We detected increased total peroxidase activity, and staining indicating increased lignification in roots as a response to C. polonica. In L. bicolor inoculated roots there was no increase in total peroxidase activity, but an additional highly acidic peroxidase isoform appeared that was not present in healthy roots, or in roots invaded by the pathogen. Increased protease activity was detected in roots colonised by C. polonica, but little protease activity was detected in L. bicolor inoculated roots. These results suggest that the pathogen efficiently invades the roots despite the induced host defence responses, while L. bicolor suppresses or evades inducing such host responses in this experimental system.
Authors
Carl Gunnar Fossdal Nina Elisabeth Nagy Ari Hietala Harald Kvaalen Rune Slimestad Steve Woodward Halvor SolheimAbstract
No abstract has been registered
Authors
Sheng-Hong Li Nina Elisabeth Nagy Almuth Hammerbacher Paal Krokene Xue-Mei Niu Jonathan Gershenzon Bernd SchneiderAbstract
Norway spruce (Picea abies) bark contains specialized phloem parenchyma cells that swell and change their contents upon attack by the bark beetle Ips typographus and its microbial associate, the blue stain fungus Ceratocystis polonica. These cells exhibit bright autofluorescence after treatment with standard aldehyde fixatives, and so have been postulated to contain phenolic compounds. Laser microdissection of spruce bark sections combined with cryogenic NMR spectroscopy demonstrated significantly higher concentrations of the stilbene glucoside astringin in phloem parenchyma cells than in adjacent sieve cells. After infection by C. polonica, the flavonoid (+)-catechin also appeared in phloem parenchyma cells and there was a decrease in astringin content compared to cells from uninfected trees. Analysis of whole-bark extracts confirmed the results obtained from the cell extracts and revealed a significant increase in dimeric stilbene glucosides, both astringin and isorhapontin derivatives (piceasides A to H), in fungus-infected versus uninfected bark that might explain the reduction in stilbene monomers. Phloem parenchyma cells thus appear to be a principal site of phenolic accumulation in spruce bark.
Authors
Nina Elisabeth Nagy Harald Kvaalen Monica Fongen Carl Gunnar Fossdal Nicholas Clarke Halvor Solheim Ari HietalaAbstract
Pathogen challenge of tree sapwood induces the formation of reaction zones with antimicrobial properties such as elevated pH and cation content. Many fungi lower substrate pH by secreting oxalic acid, its conjugate base oxalate being a reductant as well as a chelating agent for cations. To examine the role of oxalic acid in pathogenicity of white-rot fungi, we conducted spatial quantification of oxalate, transcript levels of related fungal genes, and element concentrations in heartwood of Norway spruce challenged naturally by Heterobasidion parviporum. In the pathogen-compromised reaction zone, upregulation of an oxaloacetase gene generating oxalic acid coincided with oxalate and cation accumulation and presence of calcium oxalate crystals. The colonized inner heartwood showed trace amounts of oxalate. Moreover, fungal exposure to the reaction zone under laboratory conditions induced oxaloacetase and oxalate accumulation, whereas heartwood induced a decarboxylase gene involved in degradation of oxalate. The excess level of cations in defense xylem inactivates pathogen-secreted oxalate through precipitation and, presumably, only after cation neutralization can oxalic acid participate in lignocellulose degradation. This necessitates enhanced production of oxalic acid by H. parviporum. This study is the first to determine the true influence of white-rot fungi on oxalate crystal formation in tree xylem.
Authors
Nina Elisabeth Nagy Simon Ballance Harald Kvaalen Carl Gunnar Fossdal Halvor Solheim Ari HietalaAbstract
Heterobasidion parviporum, a common pathogenic white-rot fungus in managed Norway spruce forests in northern and central Europe, causes extensive decay columns within stem heartwood of the host tree. Infected trees combat the lateral spread of decay by bordering the heartwood with a fungistatic reaction zone characterized by elevated pH and phenol content. To examine the mode of fungal feeding in the reaction zone of mature Norway spruce trees naturally infected by H. parviporum, we conducted spatial proWling of pectin and hemicellulose composition, and established transcript levels of candidate fungal genes encoding enzymes involved in degradation of the diVerent cell wall components of wood. Colonized inner heartwood showed pectin and hemicellulose concentrations similar to those of healthy heartwood, whereas the carbohydrate proWles of compromised reaction zone, irrespective of the age of fungal activity in the tissue, indicated selective fungal utilization of galacturonic acid, arabinose, xylose and mannose. These data show that the rate of wood decay in the reaction zone is slow. While the up-regulation of genes encoding pectinases and hemicellulases preceded that of the endoglucanase gene during an early phase of fungal interaction with xylem defense, the manganese peroxidase gene showed similar transcript levels during diVerent phases of wood colonization. It seems plausible that the reaction zone components of Norway spruce interfere with both lignin degradation and the associated co-hydrolysis of hemicelluloses and pectin, resulting in a prolonged phase of selective decay.
Authors
Roman Gebauer Daniel Volařík Josef Urban Isabella Børja Nina Elisabeth Nagy Toril Eldhuset Paal KrokeneAbstract
Conifers and other trees are constantly adapting to changes in light conditions, water/nutrient supply and temperatures by physiological and morphological modifications of their foliage. However, the relationship between physiological processes and anatomical characteristics of foliage has been little explored in trees. In this study we evaluated needle structure and function in Norway spruce families exposed to different light conditions and transpiration regimes. We compared needle characteristics of sun-exposed and shaded current-year needles in a control plot and a thinned plot with 50% reduction in stand density. Whole-tree transpiration rates remained similar across plots, but increased transpiration of lower branches after thinning implies that sun-exposed needles in the thinned plot were subjected to higher water stress than sun-exposed needles in the control plot. In general, morphological and anatomical needle parameters increased with increasing tree height and light intensity. Needle width, needle cross-section area, needle stele area and needle flatness (the ratio of needle thickness to needle width) differed most between the upper and lower canopy. The parameters that were most sensitive to the altered needle water status of the upper canopy after thinning were needle thickness, needle flatness and percentage of stele area in needle area. These results show that studies comparing needle structure or function between tree species should consider not only tree height and light gradients, but also needle water status. Unaccounted for differences in needle water status may have contributed to the variable relationship between needle structure and irradiance that has been observed among conifers.
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Authors
Ari Hietala Nina Elisabeth Nagy Arne Steffenrem Harald Kvaalen Carl Gunnar Fossdal Halvor SolheimAbstract
No abstract has been registered
Abstract
Two mature clones of Norway spruce (Picea abies (L.) Karst.) shown to have different level of resistance towards inoculation of Heterobasidion parviporum were compared with respect to spatiotemporal expression of transcripts related to biosynthesis of lignin, stilbenes and other phenolic compounds in response to fungal inoculation and physical wounding. Both clones responded to H. parviporum and physical wounding at transcriptional and chemical levels. Taxifolin, detected in the resistant clone only, increased in concentration following both wounding and inoculation. Concentrations of stilbenoid glucosides were highest in the susceptible clone. Following wounding or inoculation, concentrations of these glucosides increased in the susceptible clone, and quantities of their corresponding aglycones increased dramatically in both clones close to the treatment point. Significant changes in transcription were detected over the entire lesion length for all transcripts, and only the changes in a few transcripts indicated a response to inoculation with H. parviporum differing from that caused by wounding alone. The resistant clone had higher basal concentrations of lignin (LTGA) compared to the susceptible clone; concentrations increased in both clones after wounding and wounding plus inoculation treatments, but remained consistently higher in the resistant clone, suggesting higher lignin levels in the cell walls compared to the susceptible clone. In addition, the transcript level in the same clones was also measured the following year and we saw indications of primed defences for a number of gene products likely resulting from the inoculations performed 12 months prior.
Authors
Fushi Wen Ho Hyung Woo Elizabeth A. Pierson Toril Eldhuset Carl Gunnar Fossdal Nina Elisabeth Nagy Martha C. HawesAbstract
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Abstract
In field experiments, clones of Norway spruce [Picea abies (L.) Karst.] showed different degrees of resistance against pathogenic fungi inoculated into the bark that correlate with differences in polyphenolic parenchyma (PP) cells of the bark. Cells of spruce callus cultures, particularly towards the callus surface, resemble PP cells and this study looks at changes in callus cells during infection and the relative resistance of cultures from clones of low (weak) or high (strong) resistance to fungal infection. Callus cultures, initiated from trees with different resistance, were co-inoculated with Ceratocystis polonica (Siem.) C. Moreau and Heterobasidion annosum (Fr.) Bref. Callus cells from strong clones resemble PP cells of bark tissue from strong clones, having more polyphenolic bodies, while callus cells from weak clones are more similar to PP cells from those clones, which have less extensive phenolic bodies. Callus cultures from trees with weak resistance were more quickly overgrown by both species of pathogenic fungi than cultures from trees with strong resistance. Callus cells of infected cultures showed changes similar to activated PP cells of bark, including enhanced accumulation of polyphenolics. Phenolic bodies were more numerous and more extensive (larger and denser) in callus cells of strong versus weak clones under all conditions. Thus, callus cells may perform similar functions in defense as PP cells in the bark. Callus from trees of varying resistance seem to reflect the relative resistance of the trees from which they are derived, and this study indicates that some mechanisms of resistance can be studied using callus from trees of different resistance.
Authors
Øystein Johnsen Carl Gunnar Fossdal Nina Elisabeth Nagy Jørgen A.B. Mølmann OG Dæhlen Tore SkrøppaAbstract
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Abstract
Rhizoctonia solani was frequently isolated in the Italian Alps from ursery-grown European beech (Fagus sylvatica) seedlings displaying symptoms of cotyledon rot. Koch?s postulates were verified and mode of infection of the associated isolates was investigated with light and scanning electron microscopy. Population structure of the pathogen was investigated by scoring the anastomosis reaction type in pairings between different isolates from the same seedbed. One pathogen genotype showed a large distribution area within the seedbed, this implying that the inoculum had been building up in the seedbed over a longer time period. Hyphal anastomosis tests and sequence analysis of the internal transcribed spacer (ITS) region of ribosomal DNA indicated that the pathogen belongs to AG 2-1 of R. solani. ITS sequence analysis indicates that the isolates from beech are closely related to R. solani isolates causing a disease on tulip. The striking similarities between the two diseases are discussed.
Authors
Tore Skrøppa Øystein Johnsen Carl Gunnar Fossdal Rudiger Baumann Jørgen A. Mølmann Ola G Dæhlen Nina Elisabeth Nagy G Müller-StarckAbstract
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Authors
Lars Sandved Dalen Heather Danforth John Einset Carl Gunnar Fossdal Aksel Granhus Harald Kvaalen Nina Elisabeth Nagy Paivi Liisa Rinne Linda Ripel Sissel Torre Gunnhild Søgaard Christiaan van der SchootAbstract
No abstract has been registered
Authors
Nina Elisabeth Nagy Lars Sandved Dalen D. L. Jones B. Swensen Carl Gunnar Fossdal Toril EldhusetAbstract
No abstract has been registered
Authors
Nina Elisabeth Nagy Carl Gunnar Fossdal Lars Sandved Dalen Anders Lønneborg Inger Heldal Øystein JohnsenAbstract
No abstract has been registered
Authors
Nina Elisabeth Nagy Carl Gunnar Fossdal Paal Krokene Trygve Krekling Anders Lönneberg Anders Lønneborg Halvor SolheimAbstract
Polyphenolic parenchyma cells (PP cells) in Norway spruce (Picea abies (L.) Karst.) stem phloem play important roles in constitutive and inducible defenses. To determine whether anatomical and molecular changes in PP cells are correlated with tree resistance, we infected two Norway spruce clones with the pathogenic fungus Ceratocystis polonica (Siem.) C. Moreau. The fungus induced significantly different lesion lengths in the two clones, indicating that one clone was more resistant to the fungus (short lesions) than the other (long lesions). After infection, the cross-sectional area of PP cells and their vacuolar polyphenol bodies increased in the three most recent annual rings of PP cells in both clones. The more resistant clone had larger PP cells with denser polyphenol bodies than the less resistant clone, whereas the less resistant clone accumulated relatively more polyphenols after infection. Compared with the less resistant clone, the more resistant clone contained higher starch concentrations before infection that were reduced more quickly after infection before returning to original values. Low transcript levels of chalcone synthase were detected in uninfected tissues of both clones, but the levels increased dramatically after infection. Transcript levels were higher and peaked 6 days earlier in the more resistant clone than in the less resistant clone. The activity of at least one highly basic peroxidase isoform was greatly enhanced after infection, and this increase occurred earlier in the more resistant clone.
Authors
Carl Gunnar Fossdal Øystein Johnsen Nina Elisabeth Nagy R Baumann Jørgen A.B. Mølmann Tore SkrøppaAbstract
Introduction: Survival and competitive successes of boreal forest trees depend on a balance between exploiting the full growing season and minimising frost injury through proper timing of hardening in autumn and dehardening in spring. Our research indicates that the female parents of Norway spruce adjust these timing events in their progeny according to the prevailing temperature conditions during sexual reproduction. Reproduction in a cold environment advances bud-set and cold acclimation in the autumn and dehardening and flushing in spring, whereas a warm reproductive environment delays these progeny traits by an unknown non-Mendelian mechanism. We are now looking for molecular mechanisms that can explain this “epigenetic” phenomenon. Material and methods: We have performed identical crosses with the same Norway spruce (Picea abies) parent, as discussed by Skrøppa & Johnsen (1994) and Johnsen et al. (1995), in combination with timed temperature treatments during shorter and longer periods from female meiosis, pollen tube growth, syngamy and embryogenesis and tested the progenies for bud-set and frost hardiness. We have followed the transcription of the spruce phytochromes PHYO, PHYP and PHYN and the class IV chitinase PaChi4 using Quantitative Multiplex Real-Time PCR. Results and conclusions: The effect of temperature on Adaptive properties is most likely a response to accumulated heat during embryogenesis and seed maturation. Our first attempt to look for a molecular mechanism has revealed that transcription of PHYO, PHYP and PHYN and the class IV chitinase PaChi4 (relative to alphaTubulin) all show higher transcription levels in progenies born under cold conditions than their full-sibs born under warmer conditions. This result is consistent with preliminary findings that methylation of cytosine in total DNA is higher in progenies reproduce under warm conditions than their colder full-sib counterparts. If these observations are related to methylation or other epigenetic effects, we may explain why progenies with a memory of a past time cold embryogenesis are more sensitive to short days than their full-sibs with a warmer embryonic history.
Authors
Carl Gunnar Fossdal Øystein Johnsen Rüdiger Baumann Jørgen A. Mølmann Ola Gram Dæhlen Nina Elisabeth Nagy Tore SkrøppaAbstract
Research indicate that the female parents of Norway spruce adjust these timing events in their progeny according to the prevailing temperature conditions during seed development. Reproduction in a cold environment advances bud-set and cold acclimation in the autumn and dehardening and flushing in spring, whereas a warm reproductive environment delays these progeny traits by an unknown non-Mendelian mechanism. We have performed identical crosses in combination with timed temperature treatments during shorter and longer periods from female meiosis, pollen tube growth, syngamy and embryogenesis, tested the progenies for bud-set and frost hardiness, and concluded that the effect of temperature most likely is a response to accumulated heat during embryogenesis and seed maturation. Our first attempt to look for a molecular mechanism has revealed that transcription of PHYO, PHYP and PHYN and the class IV chitinase PaChi4 (using RealTime PCR) all show higher transcription levels in progenies born under cold conditions than their full-sibs born under warmer conditions. This result is consistent with preliminary findings that methylation of cytosine in total DNA is higher in progenies reproduce under warm conditions than their colder full-sib counterparts. If these observations are related to methylation, we may explain why progenies with a memory of a past time cold embryogenesis are more sensitive to short days than their full-sibs with a warmer embryonic history.
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Authors
Erik Christiansen Alan A. Berryman Vincent R. Franceschi Trygve Krekling Paal Krokene Nina Elisabeth Nagy Halvor SolheimAbstract
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Authors
Erik Christiansen V. R. Franceschi Nina Elisabeth Nagy Trygve Krekling A. A. Berryman Paal Krokene Halvor Solheim F. Lieutier W. J. Mattson M. R. WagnerAbstract
No abstract has been registered
Authors
Nina Elisabeth Nagy Kirsten Bjørklund Holven Norbert Roos Haruki Senoo Naosuke Kojima Kaare Reidar Norum Rune BlomhoffAbstract
No abstract has been registered
