Ari Hietala
Research Professor
(+47) 480 28 268
ari.hietala@nibio.no
Place
Steinkjer
Visiting address
Innocamp Steinkjer, Skolegata 22, 7713 Steinkjer
Abstract
No abstract has been registered
Authors
Chatchai Kosawang Isabella Børja Maria 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
Bacterial diseases in woody plants are best characterized for ornamental and fruit trees and much less is known for forest trees. There are many diseases of forest trees whose etiology remains to be clarified and likely more bacterial diseases of forest trees will be discovered in the next years. An overview of the main bacterial pathogens that cause diseases in forest and ornamental trees is described in this chapter and the general differences between fungal and bacterial diseases are outlined. For bacteria pathogenic to trees, six types of diseases are described: Bacterial blight diseases, represented by Erwinia amylovora, the fireblight disease; Bacterial wilt disease, represented by Ralstonia solanacearum species complex; Root and stem galls of trees, represented by Agrobacterium tumefaciens; Wetwood disease, caused by several bacterial genera like Clostridium, Bacillus, Enterobacter, Klebsiella, and Pseudomonas, Xanthomonas and Pantoea; Bacterial scorch disease represented by Xylella fastidiosa with all its subspecies; Bacterial canker represented by Pseudomonas syringae with all its pathovars. Finally, the current diagnostic methods and specific issues related to bacteria detection, together with the main results of the scientific efforts and challenges in the genetic breeding to increase bacterial resistance of trees, are outlined.
Abstract
Pathogenic wood decay fungi such as species of Heterobasidion are some of the most serious forest pathogens in Europe, causing rot of tree boles and loss of growth, with estimated economic losses of eight hundred million euros per year. In conifers with low resinous heartwood such as species of Picea and Abies, these fungi are commonly confined to heartwood and thus external infection signs on the bark or foliage of trees are normally absent. Consequently, determining the extent of disease presence in a forest stand with field surveys is not practical for guiding forest management decisions such as optimal rotation time. Remote sensing technologies such as airborne laser scanning and aerial imagery are already used to reduce the reliance on fieldwork in forest inventories. This study aimed to use remote sensing to detect rot in spruce (Picea abies L. Karst.) forests in Norway. An airborne hyperspectral imager provided information for classifying the presence or absence of rot in a single-tree-based framework. Ground reference data showing the presence of rot were collected by harvest machine operators during the harvest of forest stands. Random forest and support vector machine algorithms were used to classify the presence and absence of rot. Results indicate a 64% overall classification accuracy for presence-absence classification of rot, although additional work remains to make the classifications usable for practical forest management.
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.
Abstract
Norway’s most common tree species, Picea abies (L.) Karst. (Norway spruce), is often infected with Heterobasidion parviporum Niemelä & Korhonen and Heterobasidion annosum (Fr.) Bref.. Because Pinus sylvestris L. (Scots pine) is less susceptible to rot, it is worth considering if converting rot-infested spruce stands to pine improves economic performance. We examined the economically optimal choice between planting Norway spruce and Scots pine for previously spruce-dominated clear-cut sites of different site indexes with initial rot levels varying from 0% to 100% of stumps on the site. While it is optimal to continue to plant Norway spruce in regions with low rot levels, shifting to Scots pine pays off when rot levels get higher. The threshold rot level for changing from Norway spruce to Scots pine increases with the site index. We present a case study demonstrating a practical method (“Precision forestry”) for determining the tree species in a stand at the pixel level when the stand is heterogeneous both in site indexes and rot levels. This method is consistent with the concept of Precision forestry, which aims to plan and execute site-specific forest management activities to improve the quality of wood products while minimising waste, increasing profits, and maintaining environmental quality. The material for the study includes data on rot levels and site indexes in 71 clear-cut stands. Compared to planting the entire stand with a single species, pixel-level optimised species selection increases the net present value in almost every stand, with average increase of approximately 6%.
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.
Authors
Elisabeth Pötzelsberger Martin M. Gossner Ludwig Beenken Anna Gazda Michal Petr Tiina Ylioja Nicola La Porta Dimitrios N. Avtzis Elodie Bay Maarten De Groot Rein Drenkhan Mihai-Leonard Duduman Rasmus Enderle Margarita Georgieva Ari Hietala Björn Hoppe Herve Jactel Kristjan Jarni Srđan Keren Zsolt Keseru Marcin Koprowski Andrej Kormuťák María Josefa Lombardero Aljona Lukjanova Vitas Marozas Edurad Mauri Maria Cristina Monteverdi Per Holm Nygaard Nikica Ogris Nicolai Olenici Christophe Orazio Bernhard Perny Glória Pinto Michael Power Radoslaw Puchalka Hans Peter Ravn Ignacio Sevillano Sophie Stroheker Paul Taylor Panagiotis Tsopelas Josef Urban Kaljo Voolma Marjana Westergren Johanna Witzell Olga Zborovska Milica ZlatkovicAbstract
For non-native tree species with an origin outside of Europe a detailed compilation of enemy species including the severity of their attack is lacking up to now. We collected information on native and non-native species attacking non-native trees, i.e. type, extent and time of first observation of damage for 23 important non-native trees in 27 European countries. Our database includes about 2300 synthesised attack records (synthesised per biotic threat, tree and country) from over 800 species. Insects (49%) and fungi (45%) are the main observed biotic threats, but also arachnids, bacteria including phytoplasmas, mammals, nematodes, plants and viruses have been recorded. This information will be valuable to identify patterns and drivers of attacks, and trees with a lower current health risk to be considered for planting. In addition, our database will provide a baseline to which future impacts on non-native tree species could be compared with and thus will allow to analyse temporal trends of impacts.
Abstract
No abstract has been registered
Abstract
The relationship between the ecological success of needle pathogens of forest trees and species richness of co-inhabiting endophytic fungi is poorly understood. One of the most dangerous foliar pathogens of pine is Dothistroma septosporum, which is a widely spread threat to northern European forests. We sampled two Pinus sylvestris sites in Estonia and two in Norway in order to analyse the relations between the abundance of D. septosporum and overall fungal richness, specific fungal species composition, time of season, needle age and position in the canopy. In both countries, the overall species richness of fungi was highest in autumn, showing a trend of increase with needle age. The overall species richness in the second-year needles in Estonia and third-year needles in Norway was similar, suggesting that a critical colonization threshold for needle shed in P. sylvestris is breached earlier in Estonia than in Norway. The fungal species richness in P. sylvestris needles was largely affected by Lophodermium conigenum. Especially in older needles, the relative abundance of L. conigenum was significantly higher in spring compared to summer or autumn. The timing of recruitment and colonization mechanisms of different foliage endophytes are shortly discussed.
Abstract
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
Christer Magnusson Ari Hietala Per Hansen Eva Solbjørg Flo Heggem Irene Rasmussen Birgit Schaller Marte Persdatter Tangvik Ben Z HellalAbstract
I 2019 års kartlegging av furuvednematoden Bursaphelenchus xylophilus i Norge ble 400 prøver tatt fra hogstavfall og vindfall av Pinus sylvestris L. med angrep av furubukk Monochamus spp. Prøvene ble tatt ut i Akershus, Buskerud, Østfold, Telemark, Aust-Agder og Vest-Agder. Prøvene som besto av flis ble inkubert ved +25oC i to uker før de ble ekstrahert med Baermanntrakt og undersøkt i mikroskop. Furuvednematoden B. xylophilus ble ikke påvist i prøvene, men den naturlig forekommende arten Bursaphelenchus mucronatus kolymensis ble oppdaget i fire prøver fra Agderfylkene. Feller med feromoner for fangst av furubukk ble satt opp i Hedmark (Elverum, Romedal, Stange og ved Geitholmsjøen), Møre og Romsdal (Kvanne) og Østfold (Fredrikstad og Vestby). I laboratoriet ble billene kuttet i biter og ekstrahert med en modifisert Baermanntrakt. Suspensjonen fra ekstraksjonene ble undersøkt i stereomikroskop for forekomst av infektive stadier av Bursaphelenchus spp.. Ingen nematoder kunne påvises i de 106 undersøkte billene. I perioden 2000 – 2019 er totalt 8123 vedprøver analysert. Flest prøver er tatt i Østfold, fulgt av Hedmark, Telemark, Buskerud og Aust-Agder. I kartleggingen 2019 ble B. mucronatus kolymensis påvist i fire av de 400 vedprøvene, tilsvarende en frekvens på 0,01 (1 %). For hele perioden 2000 - 2019 ble Bursaphelenchus mucronatus kolymensis + B. macromucronatus, oppdaget i 73 av 8123 vedprøver som gir en eteksjonsfrekvens på 0,009 (ca. 1 %). I perioden 2014-2019 har Bursaphelenchus mucronatus kolymensis blitt påvist fem av totalt 581 biller, som gir den samme frekvensen som for vedprøver. B. mucronatus kolymensis og B. macromucronatus likner på B. xylophilus i generell biologi og habitatvalg. Hvis vi antar en hypotetisk frekvens i forekomsten til B. xylophilus som er 100 ganger lavere enn for disse naturlig forekommende nematodene, dvs. 0,00009, kan det antall prøver som trengs for en påvisning av B. xylophilus med 95 % konfidensintervall estimeres til 30 801. Dette indikerer at vi i dag hypotetisk sett har nådd bare 26 % av det antall prøver som trengs for å kunne erklære Norge fri for furuvednematoden B. xylophilus.
Authors
Olalla Díaz-Yáñez Blas Mola-Yudego Volkmar Timmermann Mari Mette Tollefsrud Ari Hietala Jonas OlivaAbstract
Determining the impacts of invasive pathogens on tree mortality and growth is a difficult task, in particular in the case of species occurring naturally at low frequencies in mixed stands. In this study, we quantify such effects by comparing national forest inventory data collected before and after pathogen invasion. In Norway, Fraxinus excelsior is a minor species representing less than 1% of the trees in the forests and being attacked by the invasive pathogen Hymenoscyphus fraxineus since 2006. By studying deviations between inventories, we estimated a 74% higher-than-expected average ash mortality and a 13% slower-than-expected growth of the surviving ash trees, indicating a lack of compensation by the remaining ash. We could confidently assign mortality and growth losses to ash dieback as no mortality or growth shifts were observed for co-occurring tree species in the same plots. The mortality comparisons also show regional patterns with higher mortality in areas with the longest disease history in Norway. Considering that ash is currently mostly growing in mixed forests and that no signs of compensation were observed by the surviving ash trees, a significant habitat loss and niche replacement could be anticipated in the mid-term.
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
No abstract has been registered
Authors
Marit Almvik Marie-Pierre Pavageau Nina Elisabeth Nagy Hans Ragnar Norli Ari Hietala Sven R. Odenmarck Monica Fongen Anas KamlehAbstract
No abstract has been registered
Authors
R. Ioos P. Chrétien J. Perrault C. Jeandel C. Dutech P. Gonthier F. Sillo Ari Hietala Halvor Solheim J. HubertAbstract
Four species of the destructive forest pathogen Heterobasidion annosum sensu lato (s.l.) are present in Europe: H. annosum sensu stricto (s.s.), H. abietinum and H. parviporum are native species, while H. irregulare is a non‐native invasive species currently reported only in Italy, yet recommended for regulation throughout Europe. In this study, real‐time PCR detection tests were developed for each of the four species, which can be used simultaneously or individually thanks to probes labelled with species‐specific fluorescent dyes. The different performance criteria of each assay were evaluated, and it was determined that they were theoretically capable of detecting amounts of DNA corresponding to 311, 29 and 29 cell nuclei in H. annosum s.s., H. irregulare and H. parviporum, respectively. The specificity of each assay was assessed with a wide set of strains. Real‐time PCR tests successfully detected Heterobasidion species from 36 fruiting bodies taken from the forest, as well as from artificially inoculated or naturally infected wood samples. The multiplex real‐time PCR assays developed in this study could have practical applications both in forest management and in phytosanitary monitoring.
Authors
Hugh Morris Ari Hietala Steven Jansen Javier Ribera Sabine Rosner Khalifah A. Salmeia Francis W. M. R. SchwarzeAbstract
No abstract has been registered
Abstract
No abstract has been registered
Authors
Ari Hietala Isabella Børja Hugh B. Cross Nina Elisabeth Nagy Jørn Henrik Sønstebø Volkmar Timmermann Adam Vivian-Smith Halvor SolheimAbstract
No abstract has been registered
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No abstract has been registered
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
Louise Eriksson Johanna Boberg Thomas L. Cech Tamara Corcobado Marie-Laure Desprez-Loustau Ari Hietala Marilia Horta Jung Thomas Jung Hatice Tugba Dogmus Lehtijarvi Funda Oskay Slavtcho Slavov Halvor Solheim Jan Stenlid Jonas OlivaAbstract
Political action can reduce introductions of diseases caused by invasive forest pathogens (IPs) and public support is important for effective prevention. The public’s awareness of IP problems and the acceptability of policies aiming to combat these pathogens were surveyed in nine European countries (N = 3469). Although awareness of specific diseases (e.g., ash dieback) varied, problem awareness and policy acceptability were similar across countries. The public was positive towards policies for informational measures and stricter standards for plant production, but less positive towards restricting public access to protected areas. Multilevel models, including individual and country level variables, revealed that media exposure was positively associated with awareness of IP problems, and strengthened the link between problem awareness and policy acceptability. Results suggest that learning about IPs through the media and recognizing the associated problems increase policy acceptability. Overall, the study elaborates on the anthropogenic dimension of diseases caused by IPs.
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No abstract has been registered
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No abstract has been registered
Abstract
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
Mark McMullan Maryam Rafiqi Gemy Kaithakottil Bernardo J. Clavijo Lorelei Bilham Elizabeth Orton Lawrence Percival-Alwyn Ben J. Ward Anne Edwards Diane G.O. Saunders Gonzalo Garcia Accinelli Jonathan Wright Walter Verweij Georgios Koutsovoulos Kentaro Yoshida Tsuyoshi Hosoya Louisa Williamson Philip Jennings Renaud Ioos Claude Husson Ari Hietala Adam Vivian-Smith Halvor Solheim Dan MaClean Christine Fosker Neil Hall James K.M. Brown David Swarbreck Mark Blaxter J. Allan Downie Matthew D. ClarkAbstract
Accelerating international trade and climate change make pathogen spread an increasing concern. Hymenoscyphus fraxineus, the causal agent of ash dieback, is a fungal pathogen that has been moving across continents and hosts from Asian to European ash. Most European common ash trees (Fraxinus excelsior) are highly susceptible to H.fraxineus, although a minority (~5%) have partial resistance to dieback. Here, we assemble and annotate a H.fraxineus draft genome, which approaches chromosome scale. Pathogen genetic diversity across Europe and in Japan, reveals a strong bottleneck in Europe, though a signal of adaptive diversity remains in key host interaction genes. We find that the European population was founded by two divergent haploid individuals. Divergence between these haplotypes represents the ancestral polymorphism within a large source population. Subsequent introduction from this source would greatly increase adaptive potential of the pathogen. Thus, further introgression of H.fraxineus into Europe represents a potential threat and Europe-wide biological security measures are needed to manage this disease.
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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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.
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Ash dieback, caused by the ascomycete Hymenoscyphus fraxineus, was first observed in the eastern and southernmost Norway in 2008. Based on the age of stem bark lesions, it was concluded that the fungus had arrived to the region no later than 2006. Since 2008 the annual spread of the disease northwards along the west coast of Norway has been monitored. The registration was done each year during early summer around a disease frontier recorded in the previous year. The occurrence of necrotic bark lesions in the previous-year shoots and dieback of these shoots, and isolation of H. fraxineus from the discoloured wood associated with necrotic bark lesions were used as signs of ash dieback. These records indicate an annual spread of ash dieback in the range between 25 km and 78 km, and a mean annual spread of 51 km. The cause of the spread is discussed.
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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.
Authors
Mari Mette Tollefsrud Tor Myking Jørn Henrik Sønstebø Vaidotas Lygis Ari Hietala Myriam HeuertzAbstract
During post glacial colonization, loss of genetic diversity due to leading edge effects may be attenuated in forest trees because of their prolonged juvenile phase, allowing many migrants to reach the colonizing front before populations become reproductive. The northern range margins of temperate tree taxa in Europe are particularly suitable to study the genetic processes that follow colonization because they have been little affected by northern refugia. Here we examined how post glacial range dynamics have shaped the genetic structure of common ash (Fraxinus excelsior L.) in its northern range compared to its central range in Europe. We used four chloroplast and six nuclear microsatellites to screen 42 populations (1099 trees), half of which corresponded to newly sampled populations in the northern range and half of which represented reference populations from the central range obtained from previously studies. We found that northern range populations of common ash have the same chloroplast haplotypes as south-eastern European populations, suggesting that colonization of the northern range took place along a single migration route, a result confirmed by the structure at the nuclear microsatellites. Along this route, diversity strongly decreased only in the northern range, concomitantly with increasing population differentiation and complex population substructures, a pattern consistent with a leading edge colonization model. Our study highlights that while diversity is maintained in the central range of common ash due to broad colonizing fronts and high levels of gene flow, it profoundly decreases in the northern range, where colonization was unidirectional and probably involved repeated founder events and population fluctuations. Currently, common ash is threatened by ash dieback, and our results on northern populations will be valuable for developing gene conservation strategies.
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R. Drenkhan Halvor Solheim A. Bogacheva T. Riit K. Adamson T. Drenkhan T. Maaten Ari HietalaAbstract
Dieback of European ash was first observed in Europe in the early 1990s. The disease is caused by the invasive ascomycete Hymenoscyphus fraxineus, proposed to originate from Far East Asia, where it has been considered a harmless saprotroph. This study investigates the occurrence of H. fraxineus in tissues of local ash species in the Russian Far East, and assesses its population-specific genetic variation by ITS sequencing. Shoot dieback symptoms, characteristic of H. fraxineus infection on European ash, were common, but not abundant, on Fraxinus mandshurica and Fraxinus rhynchophylla trees in Far East Russia. High levels of pathogen DNA were associated with necrotic leaf tissues of these ash species, indicating that the local H. fraxineus population is pathogenic to their leaves. However, the low levels of H. fraxineus DNA detected in shoots with symptoms, the failure to isolate this fungus from such tissues, and the presence of other fungi with pathogenic potential in shoots with symptoms indicate that local H. fraxineus strains may not be responsible (or their role is negligible) for the observed ash shoot dieback symptoms in the region. Conspicuous differences in ITS rDNA sequences detected between H. fraxineus isolates from Russian Far East and European populations suggest that the current ash dieback epidemic in Europe might not directly originate from the Russian Far East. Revision of the herbarium material shows that the earliest specimen of H. fraxineus was collected in 1962 from the Russian Far East and the oldest H. fraxineus specimen of China was collected in 2004.
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Ari HietalaAbstract
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Ari HietalaAbstract
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Nicholas Clarke Hugh Cross Toril Drabløs Eldhuset Kjersti Holt Hanssen Ari Hietala O. Janne Kjønaas Holger Lange Jørn-Frode Nordbakken Tonje Ingeborg Økland Ingvald Røsberg Silje SkårAbstract
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In Europe, common ash (Fraxinus excelsior) is being decimated because of the invasive fungus Hymenoscyphus fraxineus. In its native range in Asia this ascomycete is considered a harmless leaf associate of F. mandshurica and F. chinensis subsp. rhynchophylla. Field observations from Europe suggest that there is speciesspecific variation in disease susceptibility among European and North American Fraxinus species, but a wider comparison at the genus level has been missing so far. We assessed disease symptoms and pathogen apothecium development in 17 Fraxinus species from Asia, Europe and North America exposed to high infection pressure in a Danish arboretum. We also tested their susceptibility to pathogen infection through controlled stem and leaf inoculations and subsequently assessed the level of pathogen DNA by a qPCR assay. The results suggested the presence of a phylogenetic signal in disease susceptibility where closely related Asian, European and North American species in section Fraxinus had relatively high levels of H. fraxineus DNA in the leaves and supported high production of apothecia. Leaves from some North American species also contained relatively high levels of H. fraxineus DNA, supported moderate production of apothecia and developed lesions—stating the need to avoid introduction of H. fraxineus to North America.
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Ari HietalaAbstract
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Globally, billions of tons of carbon sequestered in trees are annually recycled back to the atmosphere through wood decomposition by microbes. In Norway, every fifth Norway spruce shows at final harvest infection by pathogenic white-rot fungi in the genera Heterobasidion and Armillaria. As these fungi can mineralize all components of wood, we predicted that they have a significant carbon footprint. Gas samples taken from infected stems were analyzed for CO2 and CH4 concentrations, and wood samples from different parts of the decay columns were incubated under hypoxic (4% O2) and anoxic laboratory conditions. In spring and summer the stem concentrations of CO2 were generally two times higher in trees with heartwood decay than in healthy trees. For most of the healthy trees and trees with heartwood decay, mean stem concentrations of CH4 were comparable to ambient air, and only some Armillaria infected trees showed moderately elevated CH4. Consistently, low CH4 production potentials were recorded in the laboratory experiment. Up-scaling of CO2 efflux due to wood decay in living trees suggests that the balance between carbon sequestration and emission may be substantially influenced in stands with high frequency of advanced root and stem heartwood decay.
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Nicola La Porta Gaurav Sablok Giovanni Emilliani Ari Hietala Alessio Giovannelli Paolo Fontana Emilio Potenza Paolo BaldiAbstract
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Ari Hietala Isabella Børja Hugh Cross Volkmar Timmermann Nina Elisabeth Nagy Halvor Solheim Jørn Henrik Sønstebø adam vivian smithAbstract
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Ari Hietala Isabella Børja Hugh Cross Nina Elisabeth Nagy Halvor Solheim Jørn Henrik Sønstebø Volkmar Timmermann Adam Vivian-SmithAbstract
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Ari Hietala Hugh Cross Isabella Børja Nina Elisabeth Nagy Volkmar Timmermann Halvor Solheim Jørn Henrik Sønstebø Adam Vivian-SmithAbstract
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O. Janne Kjønaas Nicholas Clark Toril Drabløs Eldhuset Ari Hietala Hugh Cross Kjersti Holt Hansen Tonje Ingeborg Økland Holger Lange Jørn-Frode Nordbakken Ingvald RøsbergAbstract
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O. Janne Kjønaas Nicholas Clarke Toril Drabløs Eldhuset Ari Hietala Hugh Cross Kjersti Holt Hanssen Tonje Ingeborg Økland Holger Lange Jørn-Frode Nordbakken Ingvald RøsbergAbstract
Tree harvest and different harvesting methods may affect the soil carbon (C) pool in forest ecosystems. In con- ventional stem-only timber harvesting (SOH), branches and tops that are left in the forests may contribute to the build-up of the soil carbon pool. In whole-tree harvesting (WTH), inputs of organic matter from branches and tops are strongly reduced. We established field experiments at Gaupen, SE and Vindberg, SW Norway, to study the short-term effects of SOH and WTH on processes affecting the accumulation and loss of soil C. Logging residues on the WTH plots were collected in piles that were removed after 6 months, rendering two sub treatments (WTH- pile and WTH-removal areas). We weighed selected trees and logging residues, surveyed understorey biomass production, quantified pre-harvest soil C and nutrient pools down to 30 cm. Soil respiration was measured and soil water sampled monthly during the growing season, while temperature and moisture were measured continuously. Organic and mineral horizons were incubated at different temperatures to estimate potential C and N mineraliza- tion, and deep sequencing of the ITS2 barcode region of fungal DNA was performed on the samples. Litterbags were deployed in the SOH plots. The logging residues amounted to 2.2-2.4 kg C m-2 At Gaupen, the mean in situ soil respiration rates increased following harvest with all treatments, but were significantly higher in WTH-pile and SOH relative to the WTH- removal areas in the first year as well as the fourth year of treatment. The former rates included aboveground decomposing needles and twigs but excluded coarser branches. The observed increase in the WTH-removal areas may be related to decomposing roots, as well as to increased C mineralization partly due to the higher soil tem- peratures following harvest. Soil temperature was the single most important factor explaining the variability in soil respiration rates over all treatments. At Vindberg, a decrease in soil respiration was observed with all treatments in the second and third years following harvest. At both sites, decomposition of logging residues from needles was more rapid relative to twigs and fine roots. The decomposing residues released a substantial amount of nitrogen which was gradually reflected in the soil water at 30 cm soil depth. A considerable increase in the NO3-N concen- tration also in the WTH-removal areas in the second year following harvest suggests an increase in N availability from decomposing fine roots and/or soil organic matter. The increased N availability in the WTH-removal areas was supported by results from short term lab incubations of undisturbed soil from the forest floor. The changes in the WTH-removal areas were also reflected in the soil fungal diversity: saprophytic ascomycetes on decaying plant material showed a striking increase in all treatments. For the WTH-removal areas, this may, again, be related to the increased input of root litter; however, the decrease in mycorrhizal basidiomycete species and the vigorous increase of ascomycetes following harvest may also affect the C mineralization of soil organic matter.
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
Jonas Oliva S. Rommel Carl Gunnar Fossdal Ari Hietala M. Nemesio-Gorriz Halvor Solheim Malin ElfstrandAbstract
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Authors
Ari HietalaAbstract
Polymerase chain reaction (PCR), a biochemical technology that generates copies of a particular DNA sequence, has revolutionized life sciences. Thirty years after its development in 1983, PCR is a standard and indispensable technique in medical and biological research for a variety of applications. Compared to many other fields, novel PCR applications and sequencing platforms have been rather slowly adopted by research groups engaged in wood protection. Regarding laboratory and field experiments for testing the efficacy of wood treatments, this opinion article discusses the potential of 2nd generation PCR applications and sequencing platforms for profiling 1) the growth and activity of wood decay microbes upon feeding and detoxification of treated wood, and 2) for recording the successional changes that take place in microbial community along with progress of wood decay.
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Ari HietalaAbstract
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Ari M. Hietala, Volkmar Timmermann, Isabella Børja & Halvor Solheim Norwegian Forest and Landscape Institute. PO Box 115, 1431 Ås, Norway: ari.hietala@skogoglandskap.no Owing to the Gulf Stream, the northernmost European populations of several tree species are found in Norway. Common ash (Fraxinus excelsior), the only native ash species in Norway, is present in the lowlands in the southeastern part with continental climate and in southern and southwestern coastal regions with North Atlantic climate up to Central Norway. The current standing volume of ash in Norway is ca 3 mill m3 (broadleaved trees in total 220 mill m3). The first documentation of Ash Dieback (ADB) is from 2008 from a nursery in the southeastern part of the country. A survey later that year showed that dieback symptoms were present over a distance of nearly 400 km in the southeastern region. In addition to nurseries and forests, ADB symptoms were observed on roadside, alley, garden and park trees. Based on the presence of old ADB-like stem lesions detected in 2008, the pathogen must have arrived to Norway no later than 2006. In 2008, the Norwegian Food Safety Authority laid down regulations with the aim of preventing further spread of ADB. These regulations divide the country into quarantine, observation and infection-free zones, and prohibit the export of ash seedlings, seed and wood from the quarantine zone. Despite of these regulations, the disease spread rapidly along the western coast in the period between 2009 and 2013, and currently only the ash stands in Central Norway are free of the disease. The rapid spread of the disease in Norway is obviously due to airborne dispersal of pathogen ascospores. In our experimental stand in SE Norway the number of pathogen fruit bodies can be as high as 10,000 per m2 in the peak season, mid-July to mid-August. During the early morning hours the amount of pathogen ascospores at a diseased stand can exceed 100,000 ascospores per m3 air. The first symptoms of the disease, necrotic lesions on leaf blade and petiole, appear typically during the first two weeks of August in SE Norway. To observe long-term impacts of ADB, eight monitoring plots have been established in continental and North Atlantic climate zones. In SE Norway with the oldest disease history, above 60 % of the trees with a breast height diameter (BHD) below 12.5 cm have so far died or suffer from severe defoliation, 1/3 of the larger trees being affected to a similar degree. The proportions of healthy (no signs of defoliation) small and larger trees are 20% and 37%, respectively. In SW Norway with more recent disease history a similar trend is observed but the proportion of dead trees is still small. As a consequence of ADB, the Norwegian nurseries no longer grow ash seedlings. There are currently no practical control options for the disease in forestland. Several European countries have reported that even at heavily diseased ash stands there are often some ash trees that show little symptoms. This may be due to genetic variation between trees in disease resistance, a hypothesis that is currently being investigated in several European projects. Thus implementation of forest management practices that eliminate ash could have a negative effect as survival of the tree ultimately depends on selection of trees with increased disease resistance. Bibliography for Ari M. Hietala Ari M. Hietala is a Senior Forest Pathologist at the Norwegian Forest and Landscape Institute, which is a primarily government funded organisation providing scientific research and services to government, non-governmental and commercial organisations. He has worked with a range of fungal root and shoot diseases occurring on broadleaved trees and conifers indigenous to the Nordic countries. Ari and the rest of the group participate currently in several European consortia engaged in ash dieback research.
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Authors
Nicholas Clarke Toril Drabløs Eldhuset Kjersti Holt Hanssen Ari Hietala O. Janne Kjønaas Holger Lange Jørn-Frode Nordbakken Tonje Ingeborg Økland Ingvald RøsbergAbstract
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Authors
Nicholas Clarke Toril Drabløs Eldhuset Kjersti Holt Hanssen Ari Hietala O. Janne Kjønaas Holger Lange Jørn-Frode Nordbakken Tonje Ingeborg Økland Ingvald RøsbergAbstract
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Authors
O. Janne Kjønaas Nicholas Clarke Toril Drabløs Eldhuset Kjersti Holt Hanssen Ari Hietala Holger Lange Jørn-Frode Nordbakken Tonje Ingeborg Økland Ingvald RøsbergAbstract
No abstract has been registered
Authors
O. Janne Kjønaas Nicholas Clarke Toril Drabløs Eldhuset Ari Hietala Hugh Cross Tonje Ingeborg Økland Jørn-Frode Nordbakken Holger Lange Ingvald Røsberg Kjersti Holt HanssenAbstract
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Authors
Nina Elisabeth Nagy Katarzyna Sikora Paal Krokene Ari Hietala Halvor Solheim Carl Gunnar FossdalAbstract
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Authors
Igor A. Yakovlev Ari Hietala Pierre-Emmanuel Courty Taina Lundell Halvor Solheim Carl Gunnar FossdalAbstract
The pathogenic white-rot basidiomycete Heterobasidion irregulare is able to remove lignin and hemicellulose prior to cellulose during the colonization of root and stem xylem of conifer and broadleaf trees. We identified and followed the regulation of expression of genes belonging to families encoding ligninolytic enzymes. In comparison with typical white-rot fungi, the H. irregulare genome has exclusively the short-manganese peroxidase type encoding genes (6 short-MnPs) and thereby a slight contraction in the pool of class II heme-containing peroxidases, but an expansion of the MCO laccases with 17 gene models. Furthermore, the genome shows a versatile set of other oxidoreductase genes putatively involved in lignin oxidation and conversion, including 5 glyoxal oxidases, 19 quinone-oxidoreductases and 12 aryl-alcohol oxidases. Their genetic multiplicity and gene-specific regulation patterns on cultures based on defined lignin, cellulose or Norway spruce lignocellulose substrates suggest divergent specificities and physiological roles for these enzymes. While the short-MnP encoding genes showed similar transcript levels upon fungal growth on heartwood and reaction zone (RZ), a xylem defense tissue rich in phenolic compounds unique to trees, a subset of laccases showed higher gene expression in the RZ cultures. In contrast, other oxidoreductases depending on initial MnP activity showed generally lower transcript levels on RZ than on heartwood. These data suggest that the rate of fungal oxidative conversion of xylem lignin differs between spruce RZ and heartwood. It is conceivable that in RZ part of the oxidoreductase activities of laccases are related to the detoxification of phenolic compounds involved in host-defense. Expression of the several short-MnP enzymes indicated an important role for these enzymes in effective delignification of wood by H. irregulare.
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
Shoot dieback disease of European ash caused by the ascomycete Hymenoscyphus pseudoalbidus threatens ash on a continental scale. A spore sampler placed in a diseased ash forest in Southern Norway, coupled with microscopy and DNA-based fungal species-specific real-time PCR assays, was employed to profile diurnal and within-season variation in infection pressure by ascospores of H. pseudoalbidus and the potentially co-existing non-pathogenic Hymenoscyphusalbidus. Hymenoscyphus pseudoalbidus was found to be predominant in the stand. Massive simultaneous liberation, by active discharge of pathogen ascospores in the morning, peaked in mid-Jul. to mid-Aug. Accumulation of pathogen DNA on leaflets of current-year leaves reached a high level plateau phase before appearance of autumn coloration, suggesting that pathogen establishment in leaves is terminated before the onset of leaf senescence.
Abstract
The relative frequency of Therrya fuckelii and T. pini fruiting on dead branches of Scots pine was investigated in southern Norway by examining lightning-damaged and wind-fallen trees, randomly collected branches and Nordic herbarium collections of these ascomycetes representing the order Rhytismatales. Ascus, ascospore, and subhymenium characteristics were used as criteria for species identification, while a sequence analysis of ITS rDNA gene cluster was performed to compare the relatedness of the species to each other and to corresponding fungal sequences available at the NCBI GenBank Sequence Database. In a few cases, the two Therrya species co-occurred on the same branch, but in general, whether field or herbarium material, T. fuckelii was clearly more common than T. pini.Within the Nordic countries, both species occurred throughout the natural distribution area of Scots pine. The ITS rDNA sequence of T. pini strains was 91% similar to T. fuckelii strains, the differences locating both within the internal transcribed spacers ITS1 and ITS2 and the 5.8 S rDNA gene. More variation in the ITS1-5.8S-ITS2 sequence was observed among T. pini than T. fuckelii samples; genetic implications of this finding are discussed. Upon sequence analysis, we discovered that a T. pini sequence has been deposited in the NCBI GenBank under a false identity. We emphasize the importance of co-examining strains that originate from mature fruit bodies with fully developed morphologic features as reference samples.
Authors
Carl Gunnar Fossdal Nina Elisabeth Nagy Ari Hietala Harald Kvaalen Rune Slimestad Steve Woodward Halvor SolheimAbstract
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Authors
Åke Olson Andrea Aerts Fred Asiegbu Lassaad Belbahri Ourdia Bouzid Anders Broberg Björn Canback Pedro M. Coutinho Dan Cullen Kerstin Dalman Giuliana Deflorio Linda T.A. van Diepen Christophe Dunand Sébastien Duplessis Mikael Durling Paolo Gonthier Jane Grimwood Carl Gunnar Fossdal David Hansson Bernard Henrissat Ari Hietala Kajsa Himmelstrand Dirk Hoffmeister Nils Högberg Timothy Y. James Magnus Karlsson Annegret Kohler Ursula Kües Yong-Hwan Lee Yao-Cheng Lin Mårten Lind Erika Lindquist Vincent Lombard Susan Lucas Karl Lundén Emmanuelle Morin Claude Murat Jongsun Park Tommaso Raffaello Pierre Rouzé Asaf Salamov Jeremy Schmutz Halvor Solheim Jerry Ståhlberg Heriberto Vélëz Ronald P. de Vries Ad Wiebenga Steve Woodward Igor A. Yakovlev Matteo Garbelotto Francis Martin Igor V. Grigoriev Jan StenlidAbstract
• Parasitism and saprotrophic wood decay are two fungal strategies fundamental for succession and nutrient cycling in forest ecosystems. An opportunity to assess the trade-off between these strategies is provided by the forest pathogen and wood decayer Heterobasidion annosum sensu lato. • We report the annotated genome sequence and transcript profiling, as well as the quantitative trait loci mapping, of one member of the species complex: H. irregulare. Quantitative trait loci critical for pathogenicity, and rich in transposable elements, orphan and secreted genes, were identified. • A wide range of cellulose-degrading enzymes are expressed during wood decay. By contrast, pathogenic interaction between H. irregulare and pine engages fewer carbohydrate-active enzymes, but involves an increase in pectinolytic enzymes, transcription modules for oxidative stress and secondary metabolite production. • Our results show a trade-off in terms of constrained carbohydrate decomposition and membrane transport capacity during interaction with living hosts. Our findings establish that saprotrophic wood decay and necrotrophic parasitism involve two distinct, yet overlapping, processes.
Authors
Igor A. Yakovlev Gustav Vaaje-Kolstad Ari Hietala Emil Stefanczyk Halvor Solheim Carl Gunnar FossdalAbstract
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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.
Abstract
Dieback of European ash (Fraxinus excelsior), caused by the ascomycete Hymenoscyphus pseudoalbidus (anamorph Chalara fraxinea), started around 1992 in Poland and has since then spread over large geographical areas. By November 2010, the disease had been recorded in 22 European countries. The gradual expansion and high intensity of the ash dieback epidemic in Europe may suggest that H. pseudoalbidus is an invasive alien organism. In Norway, ash dieback was first reported in spring 2008, and a survey in early summer of the same year revealed that the disease had spread over large parts of the southern and eastern regions of the country. The distance from the southernmost to the northernmost infected stands was, at that time, about 400 km. Some old necrotic lesions were also observed, indicating that the ash dieback pathogen is likely to have been present in Norway since at least 2006. In 2009, a spore sampler was installed in a diseased ash stand at Ås, South-Eastern Norway. Sampling started in late July and continued until late September. Large numbers of ascospores resembling those of H. pseudoalbidus were observed, with the maximum number of spores occurring from the end of July to mid-August. The deposition of ascospores occurred mainly between 6 and 8 a.m. Ascospores are most likely to be the primary source initiating host infections and responsible for the rapid recent spread of H. pseudoalbidus in Europe.
Abstract
Twenty cypress accessions were tested for freezing tolerance. After freezing to −15°C, differences among cypress accessions were tested by measuring electrolyte leakage and chlorophyll fluorescence. Based on these data, cypress accessions showing contrasting freezing tolerance were subjected to transcript profiling of candidate genes upon the development of cold hardening, with the ultimate goal of providing a scientific basis for selecting/breeding cypress genotypes with higher tolerance to low temperature. Nine different cypress genes were selected: a heat shock protein, a putative chaperonin, a chlorophyll-binding protein, a serine/threonine protein kinase, a putative exonuclease, a dehydrin, and three senescenceassociated proteins. Transcript levels of these genes were profiled during cold hardening under controlled conditions using real-time reverse-transcription-polymerase chain reaction. While the genes showed regulation patterns common to both cypress accessions, in the case of chaperonin, exonuclease, and some senescence-associated proteins, clonal differences in gene regulation were found. The potential relationship of these differences with cold tolerance is discussed.
Abstract
A rapid increase in the frequency of Dutch elm disease (DED), a wilting disease of elm trees caused by bark-beetle vectored fungi, was observed in the early 1990s on several wych elm stands around Oslofjord, southern Norway. To examine the current status of the disease and its impacts on elm population, disease frequency and size distribution of elms were recorded at four locations. Northern parts of Lier, a municipality most affected by DED in Norway 15 years ago, showed in the survey season 4% disease frequency, whereas 13.8% of trees were dead, the dead trees having accumulated over several years in the unmanaged stands. In southern parts of the municipality the mean disease and death percentages were 1.9 and 2.4%. Compatible with their low disease incidence in early 1990s, the other two areas now examined, municipality of Larvik and district of Grenland, showed comparably low frequency of DED. Northern part of Lier showed significantly higher overall density of elm trees per hectare than the other examined areas, and also the small elms below 5 cm in d.b.h. were most frequent in this region. In contrast, the density of large trees was lower in northern Lier than in the other examined areas. These data suggest that regeneration of the tree is not prohibited owing to the disease but that the large trees have been locally reduced in frequency as a result of DED. The superior general density of elm trees in northern Lier, owing to the exceptionally rich soil in the warm southern slopes of the region,> may have contributed to the rapid increase of DED in the area 15 years ago and to the subsequent settlement of the disease outbreak as a chronic stage.
Abstract
The causative agent of dieback on European ash (Fraxinus excelsior) was first described as Chalara fraxinea based on cultural morphology because no sexual stage of the fungus was known. Later, based on culturing of ascospores of a candidate teleomorph, morphological comparison and nuclear ribosomal internal transcribed spacer sequencing, the sexual stage of C. fraxinea was assigned as Hymenoscyphus albidus, a native and widespread species in Europe. Recently, the morphological species concept of H. albidus was shown to cover two species that cannot be separated from each other based on teleomorph characters, but which can be distinguished by several DNA markers. As a result, the strains causing ash dieback were reassigned as H. pseudoalbidus. The closely related H. albidus is presumably a non-pathogenic endophyte, but pathogenicity tests to confirm this hypothesis have not yet been performed. Genotyping of herbarium specimens has shown that H. pseudoalbidus was present in Switzerland for at least a decade prior to the epidemic outbreak in Europe. The origin of the ash dieback pathogen, and the general importance of correct pathogen identification to development of effective disease control, are discussed.
Abstract
No abstract has been registered
Abstract
Traditional wood preservatives based on biocides are effective against wood-deteriorating organisms because of their toxicity. By contrast, modified woods are non-toxic by definition. To investigate the efficiency of various wood modifications, quantitative real-time polymerase chain reaction (qPCR) was used to profile the DNA amounts of the white-rot fungus Trametes versicolor (L.) [Lloyd strain CTB 863 A] during an 8-week-long growth period in treated Pinus sylvestris (L.) sapwood. The studied wood was modified by acetylation, furfurylation, and thermal treatment. The traditional wood preservatives bis-(N-cyclohexyldiazeniumdioxy)-copper (Cu-HDO) and chromated copper arsenate (CCA) were used as references, whereas untreated P. sylvestris (L.) sapwood served as a control. The maximum levels of fungal DNA in native wood occurred at the end of the experiment. For all wood treatments, the maximum fungal DNA level was recorded after an incubation period of 2 weeks, followed by a decline until the end of the trial. For the preservative-treated woods, Cu-HDO showed the lowest level of fungal DNA throughout the experiment, indicating that exploratory hyphal growth is limited owing to the phytotoxicity of the treatment. The other treatments did not inhibit the exploratory hyphal growth phase. We conclude that qPCR studies of hyphal growth patterns within wood should provide a powerful tool for evaluating and further optimizing new wood protection systems.
Abstract
No abstract has been registered
Abstract
No abstract has been registered
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.
Abstract
No abstract has been registered
Abstract
The difficulty in subculturing biotrophic fungi complicates etiological studies related to the associated plant diseases. By employing internal transcribed spacer rDNA-targeted quantitative real-time polymerase chain reaction, we now show that the heteroecious rust Thekopsora areolata, commonly associated in natural conditions to sapling shoots and cones of Norway spruce and leaves of wild bird cherry, frequently infects nurserygrown seedlings of the conifer. A spatial sampling scheme was used to investigate seedlings and saplings of Norway spruce showing phloem necrosis: the highest concentration of DNA of T. areolata was recorded in the area with necrotic phloem. The separate analysis of bark and wood tissues suggested that the initial spread of the rust to healthy tissues neighboring the infection site takes place in the bark. A Phomopsis species found to coexist with T. areolata in several seedlings showed very high DNA levels in the upper part of the lesion, and even in the visually healthy proximal tissues above the lesions, which indicates that the ascomycete, most probably a secondary invader following primary infection by T. areolata, has a latent stage during early host colonization. We hypothesize that this hemibiotrophic mode of infection contributes to the successful coexistence of Phomopsis with a biotrophic rust.
Abstract
The area of wood protection is in a period of change. New tools are needed to understand the mode of action, and to further improve the new wood protection systems. A set of useful tools are found among the molecular methods. This paper presents an overview of some of the tools available, and the methods are exemplified by papers within the frame of wood protection issues. However, there is still a great unexplored potential within the field of wood protection by the use of various molecular methods. The majority of the work using molecular methods has been performed on species identification issues and within species variation. This paper lists some new promising molecular methods for wood protection issues and a presentation of a new project. The new project will help to gain some new knowledge about how the fungal decay processes are affected by different wood modification systems.
Authors
Karlsson Magnus Ari Hietala Harald Kvaalen Halvor Solheim Åke Olson Jan Stenlid Carl Gunnar FossdalAbstract
No abstract has been registered
Abstract
In spring 2002, extensive damage was recorded in southeast Norway on nursery-grown Norway spruce seedlings that had either wintered in nursery cold storage or had been planted out in autumn 2001. The damage was characterised by a top shoot dieback. Two visually distinct types of necroses were located either on the upper or lower part of the 2001-year-shoot. Isolations from the upper stem necroses rendered Gremmeniella abietina, while Phomopsis sp. was isolated mostly from the from the lower stem necroses. RAMS (random amplified microsatellites) profiling indicated that the G. abietina strains associated with diseased nursery seedlings belonged to LTT (large-tree type) ecotype, and inoculation tests confirmed their pathogenicity on Norway spruce seedlings. Phomopsis sp. was not pathogenic in inoculation tests, this implying it may be a secondary colonizer. We describe here the Gremmeniella - associated shoot dieback symptoms on Norway spruce seedlings and conclude that the unusual disease outburst was related to the Gremmeniella epidemic caused by the LTT type on large pines in 2001. The role of Phomopsis sp. in the tissue of diseased Norway spruce seedlings is yet unclear.
Abstract
No abstract has been registered
Abstract
The difficulty in sub-culturing biotrophic fungi complicates etiological studies related to the associated plant diseases. By employing species-specific ITS sequence stretches, we used real-time PCR to investigate the spatial colonization profiles of T. areolata and a co-existing Phomopsis species in seedlings and saplings of Norway spruce showing bark necrosis. There was a strong gradient in the colonization level of T. areolata DNA along the lesion length, with the highest DNA amount levels being recorded in the area with dark brown phloem. The separate analysis of bark and wood tissues indicated that the initial spread of the rust to healthy tissues neighbouring the infection site presumably takes place in the bark. A Phomopsis species co-existing together with T. areolata in several cases showed very high DNA levels in the upper part of the lesion outside the brown phloem area, and even in the visually healthy proximal tissues above the lesions. This indicates that this ascomycete has a latent stage during early colonization of Norway spruce shoots. This mode of infection most probably explains the successful co-existence of Phomopsis with a biotrophic rust, as their mutual interest would be to avoid triggering host cell death.
Abstract
The root-rot causing fungus Heterobasidion annosum can attack both spruce and pine trees and is the economically most damaging pathogen in northern European forestry. We have monitored the H. annosum S-type (fairly recently named H. parviporum) colonization rate and expression of host chitinases and other host transcripts in Norway spruce material with differing resistances using quatitative realtime PCR. Transcript levels of three chitinases, representing classes I, II and IV, were monitored. Ramets of two 33-year-old clones differing in resistance were employed as host material and inoculation and wounding was performed. clones in the area immediately adjacent to inoculation. Fourteen days after infection, pathogen colonization was restricted to the area immediately adjacent to the site of inoculation for the strong clone (589), but had progressed further into the host tissue in the weak clone (409). Transcript levels of the class II and IV chitinases increased following wounding or inoculation, while the transcript level of the class I chitinase declined following these treatments. Transcript levels of the class II and class IV chitinases were higher in areas immediately adjacent to the inoculation site in 589 than in similar sites in 409 three days after inoculation, suggesting that the clones differ in the rate of pathogen perception and host defense signal transduction. This an earlier experiments using mature spruce clones as substrate indicate that it is the speed of the host response and not maximum amplitude of the host response that is the most crucial component in an efficient defense in Norway spruce toward pathogenic fungi such as H. annosum.
Abstract
No abstract has been registered
Abstract
No abstract has been registered
Abstract
No abstract has been registered
Authors
Magnus Karlsson Ari M. Hietala Harald Kvaalen Halvor Solheim Åke Olson Jan Stenlid Carl Gunnar FossdalAbstract
Norway spruce (Picea abies (L.) Karst.) has a natural distribution in the northern parts of Europe and Asia and is economically the most important tree species grown in the Nordic countries. A common threat to Norway spruce is the basidiomyceteous fungus Heterobasidion parviporum Niemelä and Korhonen. H. parviporum mainly attacks Norway spruce, although Siberian fir (Abies sibirica Ledeb.) and Scots pine (Pinus sylvestris L.) occasionally get infected. One obstacle to studying host/pathogen interaction in conifers has been the limited availability of mature clones for controlled inoculations, as genetic variation within the host material and the lack of replicates complicate interpretation of the results. Somatic embryogenesis, rooted cuttings, and tissue cultures may provide solutions for this problem. Tissue cultures from mature Norway spruce trees have been proposed as a possible model system for assessing resistance toward fungal pathogens. Recent data on chitinase isoform activity in the Norway spruce/H. parviporum pathosystem are encouraging; clonal variation was observed in the isoforms affected by inoculation, and the isoforms showing increased band intensity following bark inoculation by H. parviporum were also induced in the inoculated tissue cultures of the corresponding clones. To investigate the biological relevance of tissue cultures in host-pathogen interaction studies, transcript levels of selected host and pathogen genes in tissue cultures of Norway spruce were compared to those in bark of 33-year-old ramets of the same clones upon challenge by the pathogenic fungus H. parviporum. Similar transcript profiles of the pathogen and host genes were observed in both tissues, this supporting the use of tissue cultures as experimental material for the pathosystem. Higher transcript levels of the host genes phenylalanine ammonia lyase, peroxidase, and glutathione-S-transferase were observed in the more resistant clone #589 than in the less resistant clone #409 during the early stages of colonization. The most striking difference between the spruce clones was related to gene transcript levels of a class IV chitinase, which showed a continuous increase in clone #409 over the experimental period, with a possible association of this gene product to programmed cell death. Several of the fungal genes assayed were differentially expressed during colonization, including putative glutathione-S-transferases, laccase, cellulase, cytochrome P450 and superoxide dismutase genes. The transcriptional responses suggest an important role for the antioxidant systems of both organisms.
Abstract
In spring 2002, extensive damage was recorded in southeast Norway on nursery-grown Norway spruce seedlings that had either wintered in nursery cold storage or had been planted out in autumn 2001. The damage was characterised by a top shoot dieback. Two visually distinct types of necroses were located either on the upper or lower part of the 2001-year-shoot. Isolations from the upper stem necroses rendered Gremmeniella abietina, while Phomopsis sp. was isolated mostly from the lower stem necroses. RAMS (random amplified microsatellites) profiling indicated that the G. abietina strains associated with diseased nursery seedlings belonged to LTT (large-tree type) ecotype,and inoculation tests confirmed their pathogenicity on Norway spruce seedlings. Phomopsis sp. was not pathogenic in inoculation tests, this implying it may be a secondary colonizer. We describe here the Gremmeniella – associated shoot dieback symptoms on Norway spruce seedlings and conclude that the unusual disease outburst was related to the Gremmeniella epidemic caused by the LTT ecotype on large Scots pines in 2001. The role of Phomopsis sp. in the tissue of diseased Norway spruce seedlings is yet unclear.
Abstract
*Strawberry Fragaria × ananassa (cv. Korona) was inoculated with Botrytis cinerea by dipping berries in a conidial suspension. *Colonization by the pathogen was monitored using real-time PCR, ELISA and ergosterol assays, the first showing the highest sensitivity. The expression of pathogen B-tubulin and six polygalacturonases (Bcpg1-6) and three host defence genes (polygalacturonase-inhibiting protein (FaPGIP) and two class II chitinases) were monitored using real-time RT-PCR. *The maximum transcript levels of the host defence genes occurred at 16 h postinoculation (hpi) at the presumed initial penetration stage. The unique transcript profile of Bcpg2 over the 96-h incubation time and its high transcript levels relative to those of the other Bcpgs at 8-24 hpi suggest that the gene has a specific role in the penetration stage. *Bcpg1 was expressed constitutively at a relatively high level in actively growing mycelia throughout the experimental period. Comparison of the transcript profiles indicated that Bcpg1 and Bcpg3-6 were coordinately regulated.
Abstract
This paper describes the use of quantitative real-time PCR for monitoring colonization of birch wood (Betula pubescens) by the white-rot fungus Trametes versicolor in an EN113 decay experiment. The wood samples were harvested after 4, 8, 12, 16 and 20 weeks of incubation.The mass loss was in the range of 440%. Chitin and ergosterol assays were conducted for comparison. Second-order polynomial fits of the mass loss of decayed wood versus chitin, ergosterol and DNA gave correlations (r2) of 0.87, 0.61 and 0.84, respectively. Compared to the other two assays employed, real-time PCR data correlated best with the relative mass loss of decayed samples 48 weeks after inoculation, while the saturation and decline of DNA-based estimates for fungal colonization 1620 weeks after inoculation indicated that the DNA assay is not suited for quantification purposes in the late stages of decay.The impact of conversion factors, extraction efficiency, inhibitory compounds and background levels in relation to the three detection assays used is discussed.
Abstract
In spring 2002, extensive damages were recorded in southeast Norway on nursery-grown Norway spruce seedlings that had either wintered in nursery cold storage or had been planted out in autumn 2001. The damages were characterised by leader shoot dieback and necroses on the upper or lower part of the 2001-year-shoot. Gremmeniella abietina and Phomopsis sp. were frequently isolated from the diseased seedlings. RAMS (random amplified microsatellites) profiling indicated that the G.abietina strains associated with diseased nursery seedlings belonged to LTT (large-tree type) ecotype, and inoculation tests confirmed their pathogenicity on Norway spruce. Based on sequence analysis of the internal transcribed spacer (ITS) regions of ribosomal DNA, the Phomopsis strains associated with diseased seedlings do not represent any characterized Phomopsis species associated with conifers. Phomopsis sp. was not pathogenic in inoculation tests, this implying it may be a secondary colonizer. ITS-based real-time PCR assays were developed in order to detect and quantify Gremmeniella and Phomopsis in the nursery stock. We describe here the Gremmeniella - associated shoot dieback symptoms on Norway spruce seedlings and conclude that the unusual disease outburst was related to the Gremmeniella epidemic caused by the LTT type on large pines in 2001.
Authors
Axel Schmidt Zeneli Gazmend Ari Hietala Carl Gunnar Fossdal Paal Krokene Erik Christiansen Jonathan GershenzonAbstract
No abstract has been registered
Authors
Magnus Karlsson Ari M. Hietala Harald Kvaalen Åke Olson Halvor Solheim Jan Stenlid Carl Gunnar FossdalAbstract
No abstract has been registered
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.
Abstract
No abstract has been registered
Abstract
Introduction: The objectives of the present study were to monitor H. annosum colonization rate (Hietala et al., 2003) and expression of host chitinases in clonal Norway spruce material with differing resistances. Transcript levels of three chitinases, representing classes I, II and IV, were monitored with real-time PCR.Material and MethodsInoculation experiment: Ramets of two 32 -year-old clones differing in resistance were employed as host material. Inoculation and wounding was performed. A rectangular strip containing phloem and cambium, with the inoculation site in the middle, was removed 3, 7 and 14 days after inoculation.Quantification of fungal colonizationMultiplex real-time PCR detection of host and pathogen DNA was performed (Hietala et al., 2003). Quantification of gene expression: Chitinase levels were monitored with Singleplex real-time PCR.Results and ConclusionsThe colonization profiles provided by the quantitative multiplex real-time PCR procedure (Hietala et al., 2003), when combined with spatial and temporal transcript profiling of 3 chitinases, provide a useful basis for identifying defense related genes, and for assessing their impact on pathogen colonization rates.Three days after inoculation, comparable colonization levels were observed in both clones in the area immediately adjacent to inoculation. Fourteen days after infection, pathogen colonization was restricted to the area immediately adjacent to the site of inoculation for the strong clone (589), but had progressed further into the host tissue in the weak (409) clone.Transcript levels of the class II and IV chitinases increased following wounding or inoculation, while the transcript level of the class I chitinase declined following these treatments. Transcript levels of the class II and class IV chitinases were higher in areas immediately adjacent to the inoculation site in 589 than in similar sites in 409 three days after inoculation, suggesting that the clones differ in the rate of chitinase-related signalperception.
Abstract
We have monitored the H. annosum colonization rate and expression of host chitinases in Norway spruce material with differing resistances. Transcript levels of three chitinases, representing classes I, II and IV, were monitored with real-time PCR. Ramets of two 32 -year-old clones differing in resistance were employed as host material and inoculation and wounding was performed. Quantification of fungal colonization: Multiplex real-time PCR detection of host and pathogen DNA was performed. Chitinase transcript levels were also monitored with real-time PCR. Three days after inoculation, comparable colonization levels were observed in both clones in the area immediately adjacent to inoculation. Fourteen days after infection, pathogen colonization was restricted to the area immediately adjacent to the site of inoculation for the strong clone (589), but had progressed further into the host tissue in the weak clone (409). Transcript levels of the class II and IV chitinases increased following wounding or inoculation, while the transcript level of the class I chitinase declined following these treatments. Transcript levels of the class II and class IV chitinases were higher in areas immediately adjacent to the inoculation site in 589 than in similar sites in 409 three days after inoculation, suggesting that the clones differ in the rate of chitinase-related signal perception. The spatiotemporal accumulation patterns obtained for the two clones used are consistent with their resistance classifications, these warranting further and more detailed studies on these chitinases.
Abstract
Pathogen colonization and transcript levels of three host chitinases,putatively representing classes I, II, and IV, were monitored with real-time PCR after wounding and bark infection by Heterobasidion annosum in 32-year-old trees of Norway spruce (Picea abies) with low (clone 409) or high (clone 589) resistance to this pathogen. Three days after inoculation, comparable colonization levels were observed in both clones in the area immediately adjacent to inoculation. At 14 days after infection, pathogen colonization was restricted to the area immediately adjacent to the site of inoculation for clone 589 but had progressed further into the host tissue in clone 409. Transcript levels of the class II and IV chitinases increased after wounding or inoculation, but the transcript level of the class I chitinase declined after these treatments. Transcript levels of the class II and class IV chitinases were higher in areas immediately adjacent to the inoculation site in clone 589 than in similar sites in clone 409 3 days after inoculation. This difference was even more pronounced 2 to 6 mm away from the inoculation point, where no infection was yet established, and suggests that the clones differ in the rate of chitinase-related signal perception or transduction. At 14 days after inoculation, these transcript levels were higher in clone 409 than in clone 589, suggesting that the massive upregulation of class II and IV chitinases after the establishment of infection comes too late to reduce or prevent pathogen colonization.
Abstract
A multiplex real-time PCR assay was developed to monitor the dynamics of the Picea abies-Heterobasidion annosum pathosystem. Tissue cultures and 32-year-old trees with low or high resistance to this pathogen were used as the host material. Probes and primers were based on a laccase gene for the pathogen and a polyubiquitin gene for the host.The real-time PCR procedure was compared to an ergosterol-based quantification method in a tissue culture experiment, and there was a strong correlation product moment correlation coefficient, 0.908) between the data sets. The multiplex real-time PCR procedure had higher resolution and sensitivity during the early stages of colonization and also could be used to monitor the host.In the tissue culture experiment, host DNA was degraded more rapidly in the clone with low resistance than in the clone with high resistance. In the field experiment, the lesions elicited were not strictly proportional to the area colonized by the pathogen.Fungal colonization was more restricted and localized in the lesion in the clone with high resistance, hereas in the clone with low resistance, the fungus could be detected until the visible end of the lesion. Thus, the real-time PCR assay gives better resolution than does the traditionally used lesion length measurement when screening host clones for resistance.
Abstract
A quantitative multiplex real-time PCR procedure was developed to monitor the dynamics in Norway spruce (Heterobasidion annosum) pathosystem. The assay reliably detected down to 1 pg of H. annosum DNA and 1 ng of host DNA in multiplex conditions. As a comparative method for quantifying fungal colonization,we applied the ergosterol assay. There was a very high correlation between the results obtained with the two methods, this strengthening the credibility of both assays. The advantages and disadvantages of these assays are discussed.
Abstract
Determining the level of pathogenic fungi and other microorganisms during colonization of the host is central in phytopathological studies. A direct way to monitor fungal hyphae within the host is microscopic examination, but chitin and ergosterol-levels are commonly used to indirectly measure the amount of fungus present. Recently real-time PCR technology is being used to follow infection agents in host tissues. We study the molecular basis of host defense responses, using the coniferous host Norway spruce infected with the pathogen Heterobasidion parviporum as the experimental system. This basidiomycete and the closely related pathogen H. annosum are the major root rot causing pathogens in conifers. To screen host material for differential resistance towards H. parviporum, it is a necessity to quantify the fungal colonization of the host tissues. Therefore, we aimed to develop and compare the sensitivity of a real-time PCR to an ergosterol based method for determining the rate of colonization, and applied the methods to rank the infection level of the pathogen on the spruce clones 053 and 589. We developed a quantitative multiplex real-time PCR procedure that reliably detecting down to 1pg H. parviporum DNA and 1ng host DNA. There was a very high correlation between the fungal-biomass/total-biomass and fungal-DNA/total-DNA rankings obtained with ergosterol and real-time PCR, strengthening the credibility of both methods. Based on both ergosterol and real-time PCR, it was clear that the clone 053 was hosting more fungal biomass than clone 589. The results indicate that this real-time procedure can be a useful method to screen different spruce material for their relative resistance to the pathogen H. parviporum.
Abstract
Determining the level of pathogenic fungi and other microorganisms during colonization of the host is central in phytopathological studies. A direct way is to monitor fungal hyphae by microscopic examination, but indirect chitin and ergosterol-based assays have been among the most applied methods in determining fungal biomass within host tissues. Recently real-time technology is increasingly receiving attention as a way to follow infection agents in host tissues.We study the molecular basis of host defense responses, using the coniferous host Norway spruce (Picea abies) infected with the basidomycete Heterobasidion annosum as the experimental system. This basidiomycete is the major root rot causing pathogens in conifers of all age classes.In order to screen host material for differential resistance towards H.annosum for both scientific and commercial reasons, it is a necessity to reliably quantify the fungal colonization of the host tissues. Therefore, the aim of this study was to develop and compare the sensitivity of a real-time PCR assay to an ergosterol based method for determining the rate of colonization by H.annosum in inoculated spruce material. We also applied the methods to rank the infection level of the pathogen on the spruce tissue culture clones.We were able to develop a quantitative multiplex real-time PCR procedure that reliably detecting down to 1pg H.annosum DNA and 1ng host DNA in DNA extracted from infected tissues. There was a very high correlation between the fungal-biomass/total-biomass and fungal DNA-total DNA rankings obtained with ergosterol and real-time PCR respectively, strengthening the credibility of both methods.Based on both ergosterol and real-time PCR, it was clear that some spruce clones were faster and more heavily infected than others. These results indicate that both ergosterol and this real-time procedure can be useful methods to screen different spruce material for their relative resistance to the pathogen H.annosum.
Abstract
One of our main interests is to learn about the molecular basis of host defense responses, using the coniferous host Norway spruce infected with the pathogen Heterobasidion parviporum as the experimental system. This basidiomycete and the closely related pathogen H. annosum are the major root rot causing pathogens in conifers.To screen host material for differential resistance towards H. parviporum, it is a necessity to quantify the fungal colonization of the host tissues. Therefore, we aimed to develop and compare the sensitivity of a real-time PCR to an ergosterol based method for determining the rate of colonization. We developed a quantitative multiplex real-time PCR procedure that reliably detecting down to 1pg H. parviporum DNA and 1ng host DNA.There was a very high correlation between the fungal-biomass/total-biomass and fungal-DNA/total-DNA rankings obtained with ergosterol and real-time PCR, strengthening the credibility of both methods. The results indicate that this real-time procedure can be a useful method to screen different spruce material for their relative resistance to the pathogen H. parviporum.