scholarly journals Mycobiome of Fraxinus excelsior With Different Phenotypic Susceptibility to Ash Dieback

2021 ◽  
Vol 4 ◽  
Author(s):  
Marta Agostinelli ◽  
Diem Nguyen ◽  
Johanna Witzell ◽  
Michelle Cleary

For the last two decades, large-scale population decline of European ash (Fraxinus excelsior) has occurred in Europe because of the introduction of the alien fungal pathogen, Hymenoscyphus fraxineus, from East Asia. Since European ash is a keystone species having critical importance for biodiversity, and only a small percentage of the ash population appears to show some tolerance against the pathogen, the loss of ash trees means that other associated organisms, especially those with high or obligate associations to ash, are at risk of further species declines. In this study, we used high throughput DNA sequencing and multivariate analysis to characterize: (i) the mycobiome in aerial tissues (i.e., leaf, bark, and xylem) of ash trees showing different phenotypic response to ash dieback, (ii) the temporal variation in fungal communities across the growing season, and (iii) the similarity in fungal community structure between ash and other common trees species that may serve as an ecological niche substitute for ash microfungi. Results showed that fungal communities differed among the three tissue types, susceptibility classes, in time and between sites. Trophic analysis of functional groups using the FUNGuild tool indicated a higher presence of pathotrophic fungi in leaves than in bark and xylem. The share of pathotrophic fungi increased along a gradient of low to high disease susceptibility in both bark and xylem tissue, while the proportion of symbiotrophic fungi correspondingly decreased in both tissue types. Neighboring, alternative host trees did not share all the fungal species found in ash, however, most microfungi uniquely associated to ash in this study are generalists and not strictly host specific. The progressive disappearance of ash trees on the landscape imposes a high risk for extinction of Red-listed macrofungal species, and breeding for resistance against ash dieback should help sustain important biodiversity associated to ash. Microfungal diversity though may be less prone to such demise since most ash-associated endophytes appear to occur on a broad range of host species.

2020 ◽  
Vol 73 (1) ◽  
pp. 43-51
Author(s):  
Dmitrii A. Shabunin ◽  
Andrey V. Selikhovkin ◽  
Elena Yu. Varentsova ◽  
Dmitry L. Musolin

Abstract The weakening and decline of European ash Fraxinus excelsior L. and other ash species have been recorded at different locations in the suburbs of Saint Petersburg, Russia. During the summer of 2019 and spring of 2020, samples from leaves, petioles, and shoots were collected from the weakened and declining ash trees in three parks in Pushkin and Gatchina and maintained in humid chambers to induce the fructification of fungi. In total, 30 taxa of micromycetes belonging to 23 genera were identified using methods of light microscopy. Hymenoscyphus fraxineus, a putative agent of ash dieback, was not recorded in the samples collected in the crowns of trees, but only on the petioles of the fallen leaves in spring. Out of all the micromycetes recorded, only coelomycetes from the genus Diplodia Fr. (in particular, D. mutila) can damage the branches of ash trees and, thus, be considered pathogenic. It is likely that H. fraxineus opens “the entry of infection” and Diplodia spp. cause the major weakening and decline of branches. The data obtained can significantly change our understanding of the causes of ash dieback and possible methods of ash stand preservation. The reason for the low pathogenicity and activity of H. fraxineus, as well as the possible role of ascomycetes Diplodia spp. in the dieback of ash stands requires further research.


2019 ◽  
Vol 25 (2) ◽  
pp. 193-202
Author(s):  
Kateryna V Davydenko ◽  
Valentyna Borysova ◽  
Olena Shcherbak ◽  
Yevhen Kryshtop ◽  
Valentyna Meshkova

The health condition of European ash (Fraxinus excelsior) stands in Ukraine has become worse since 2006. Firstly, in 2011 an alien invasive pathogenic fungus Hymenoscyphus fraxineus was identified in the eastern part of the country and, subsequently, its presence was confirmed in the western and central parts. The aims of our research were to evaluate the health of ash trees and identify the main causes of ash decline in different regions of Ukraine with emphasis on ash dieback and its association with collar rots.    Results showed that since 2013 the number of trees with ash dieback symptoms has been gradually increasing, reaching up to 92 % in 2018. Total mortality due to ash dieback was up to 9 % in 2018. Disease intensity remains high in northern and central Ukraine comparing with the east. Branch dieback, collar rots, epicormic shoots and bacterial disease of ash occurred more often in the eastern region, some symptoms were observed simultaneously. Ash bark beetle galleries, as well as foliage browsing insects, were found mostly in weakened and/or dying trees.    It was indicated that collar rots significantly increase the mortality of ash trees. Armillaria spp. fungi were found to be frequently associated with ash dieback on living stems and fallen trees in 2017, causing high rates of mortality in the northern and central regions. For further ash conservation and breeding programmes, resistant trees in severely damaged regions should be selected to preserve genetic diversity in ash populations.                                                       Keywords: Fraxinus excelsior, ash dieback, collar rot, bacterial disease, epicormic shoots, Hymenoscyphus fraxineus, Armillaria spp.


2018 ◽  
Vol 1 (1) ◽  
pp. 48-58 ◽  
Author(s):  
Tim L. R. Coker ◽  
Jiří Rozsypálek ◽  
Anne Edwards ◽  
Tony P. Harwood ◽  
Louise Butfoy ◽  
...  

Plant Disease ◽  
2021 ◽  
Author(s):  
Benedetto T. Linaldeddu ◽  
Carlo Bregant ◽  
Lucio Montecchio ◽  
Ana Brglez ◽  
Barbara Piškur ◽  
...  

Over the last decades the vitality and productivity of European ash trees in Slovenia has been seriously impacted by the onset of canker and dieback disease symptoms on young and old trees, primarily identified as ash dieback caused by Hymenoscyphus fraxineus. Given the limited information available about the aetiology of this emerging disease, a study was carried out to isolate, identify and characterize the fungal species involved in the observed ash symptoms. Field surveys were conducted in five forest sites where 50 symptomatic branch samples were collected. All samples were inspected and used for fungal isolation. Based on morphology, colony appearance and DNA sequence data of the internal transcribed spacer region (ITS), 125 fungal colonies belonging to five species were isolated and identified. Only a few symptomatic ash samples yielded colonies of H. fraxineus, whereas Botryosphaeriaceae species were isolated with a high frequency, with Diplodia fraxini as the dominant species. A pathogenicity test proved that all isolated species were pathogenic on European ash, causing bark lesions and wood discoloration. All Botryosphaeriaceae species isolated in this study are reported for the first time on European ash in Slovenia.


Forests ◽  
2020 ◽  
Vol 11 (8) ◽  
pp. 883
Author(s):  
Benedetto T. Linaldeddu ◽  
Francesco Bottecchia ◽  
Carlo Bregant ◽  
Lucia Maddau ◽  
Lucio Montecchio

In Italy, after the first report in Friuli-Venezia Giulia along the border with Slovenia in 2009, ash dieback has successively been reported in Veneto, Tuscany and Trentino-Alto Adige. Given its alarming expansion in European ash formations along the sub-montane belt of north-eastern Italy and the limited information about the associated fungal microorganisms; since 2017, a study has been conducted in order to isolate and characterize the fungal species involved in the aetiology of the disease. The surveys were conducted in six ash–maple forests distributed along the Veneto and Friuli-Venezia Giulia pre-Alpine regions (Italy). In each site, the health status of ash trees was assessed and a sample (shoot or branch) with the typical symptoms of the disease was taken from ten trees to isolate the associated pathogens. The fungal colonies developed were identified using morphological features and DNA sequences. The 60 samples processed yielded a total of 109 fungal isolates belonging to 9 families including: Botryosphaeriaceae (62 isolates), Diaporthaceae (18), Nectriaceae (10), Didymellaceae (9), Helotiaceae (5), Diatrypaceae (2), Didymosphaeriaceae (1), Phaeosphaeriaceae (1) and Valsaceae (1). In particular, three species—Diplodia subglobosa, Diplodia fraxini and Diaporthe eres—were isolated with high frequency, while Hymenoscyphus fraxineus was isolated from only five plants distributed in four sites. The pathogenicity tests, conducted on 3-year-old seedlings, detached branches (3–4 cm diameter), and leaves of Fraxinus excelsior, showed that Diplodia fraxini is the most virulent species and the only one able to reproduce the symptoms observed in nature. Overall, the results obtained emphasize that several fungal pathogens are involved in the aetiology of the disease, many of which belong to the Botryosphaeriaceae family.


2020 ◽  
Vol 71 (19) ◽  
pp. 6074-6083 ◽  
Author(s):  
Miguel Nemesio-Gorriz ◽  
Riya C Menezes ◽  
Christian Paetz ◽  
Almuth Hammerbacher ◽  
Marijke Steenackers ◽  
...  

Abstract Ash dieback, a forest epidemic caused by the invasive fungus Hymenoscyphus fraxineus, threatens ash trees throughout Europe. Within Fraxinus excelsior populations, a small proportion of genotypes show a low susceptibility to the pathogen. We compared the metabolomes from a cohort of low-susceptibility ash genotypes with a cohort of high-susceptibility ash genotypes. This revealed two significantly different chemotypes. A total of 64 candidate metabolites associated with reduced or increased susceptibility in the chemical families secoiridoids, coumarins, flavonoids, phenylethanoids, and lignans. Increased levels of two coumarins, fraxetin and esculetin, were strongly associated with reduced susceptibility to ash dieback. Both coumarins inhibited the growth of H. fraxineus in vitro when supplied at physiological concentrations, thereby validating their role as markers for low susceptibility to ash dieback. Similarly, fungal growth inhibition was observed when the methanolic bark extract of low-susceptibility ash genotypes was supplied. Our findings indicate the presence of constitutive chemical defense barriers against ash dieback in ash.


Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2524
Author(s):  
Anna Hebda ◽  
Aleksandra Liszka ◽  
Piotr Zgłobicki ◽  
Katarzyna Nawrot-Chorabik ◽  
Jan J. Lyczakowski

The population of European ash (Fraxinus excelsior L.) is currently facing the risk of collapse, mainly due to ash dieback, a disease caused by a pathogenic fungus, Hymenoscyphus fraxineus. To facilitate studies into the molecular basis of ash dieback and design breeding strategies for a generation of resistant trees, it is necessary to develop tools enabling the study of gene function in F. excelsior. Despite this, a method for the genetic engineering of F. excelsior is still missing. Here, we report the first successful genetic transformation of F. excelsior callus and a selection process enabling the formation of stable transgenic callus lines. The protocol relies on the use of Agrobacterium tumefaciens to transform callus tissue derived from embryos of F. excelsior. In our experiments, we used the β-glucuronidase (GUS) reporter system to demonstrate the transformation of callus cells and performed RT-PCR experiments to confirm the stable expression of the transgene. Since ash dieback threatens the long-term stability of many native F. excelsior populations, we hope that the transformation techniques described in this manuscript will facilitate rapid progress in uncovering the molecular basis of the disease and the validation of gene targets previously proposed to be linked to the resistance of trees to H. fraxineus pathogenicity.


2020 ◽  
Author(s):  
◽  
Ilze Matisone ◽  

The area of common ash forest is only 1–2% of the total forest area in Europe and 0.3% in Latvia. Prior to dieback, ash was an economically and still is ecologically highly important tree species. In Eastern Europe since the beginning of the 1990s, and a decade later throughout Central and Western Europe, rapid dieback of ash stands caused by an unknown reason was observed. In 2006 the causal agent of ash dieback – an anamorphic fungus Hymenoscyphus fraxineus (species name later refined) was determined. In Latvia, intensive dieback of ash stands has been observed since 2000. The aim of the thesis was to assess the dieback dynamics of common ash Fraxinus excelsior L. stands and determine the main factors affecting susceptibility of the trees, as well as future succession and potential regeneration of the affected common ash stands in Latvia. In Latvia, The area of young ash stands decreased by 4.4 times during 2000–2015. The ten-year-inventory (2005–2015) revealed that the overall mortality of ash was 6.9% and standing volume of ash stands decreased by 171 m3 ha–1. The obtained results indicated that regeneration of ash saplings in ash young stands as well as in mature stands was lower than in in the pre-dieback times, but regeneration was still successful. Furthermore, survival and health condition of ash saplings was relatively poor, as only ca. ⅓ of individuals were healthy, and the health condition decreased with aging. Overgrowth of ash stands with shrub species has begun, suggesting formation of broadleaved stands with a small admixture of ash in the future. We found that, susceptibility to ash dieback is significantly affected by morphometric parameters of ash trees and stands, site conditions and meteorological factors. In Latvia, ash dieback was present in ash stands of all ages and composition, yet some individual and regional differences were observed. Lower susceptibility to ash dieback was observed for older/larger dimensions trees, mixed stands on well-drained and dry mineral soils, stands thinned more intensively, and in the eastern part of Latvia. The findings of the thesis are applicable for the planning of ash stand management, assessment of the existing situation, for the designation of additional genetic resource forests, as well as provide basic information for future studies.


2021 ◽  
Author(s):  
Jan-Peter George ◽  
Tanja GM Sanders ◽  
Volkmar Timmermann ◽  
Nenad Potocic ◽  
Mait Lang

European ash (Fraxinus excelsior) and narrow-leafed ash (F. angustifolia) are keystone forest tree species in Europe with a broad ecological amplitude and significant economic importance. Besides global warming both species are currently under significant thread by an invasive fungal pathogen that has been progressively spreading throughout the continent for almost three decades. Ash dieback caused by the invasive ascomycete Hymenoscyphus fraxineus is capable of damaging ash trees of all age classes and often leads to the ultimate death of a tree after years of progressively developing crown defoliation. While studies at national and regional level already suggested rapid decline of ash populations as a result of ash dieback, a comprehensive survey at European level with harmonized crown assessment data across countries could shed more light into the population decline from a pan-European perspective and could also pave the way for a new conservation strategy beyond national boarders. Here, we present data from the ICP Forests Level I crown condition monitoring including 27 countries, covering the timespan from 1987-2020. In total, 407 survey plots randomly distributed across these countries were analyzed resulting in >36,000 individual observations. We found a substantial increase in defoliation and mortality over time indicating that crown defoliation has almost doubled during the last three decades. Hotspots of mortality are currently situated in southern Scandinavia and north-eastern Europe, well corresponding to the fact that the disease spread fast from north-east to north-west. Overall survival probability after nearly 30 years of infection has already reached a critical value of 0.51, but with large differences among regions (0.00-0.907). Both a Cox proportional hazard model as well as an Aalen additive regression model strongly suggest that survival of ash is significantly lower in locations with excessive water regime and which experienced more extreme precipitation events during the last two decades. Our results underpin the necessity for fast governmental acting and joint rescue efforts beyond national boarders since overall mean defoliation will likely reach 50% as early as 2030 as suggested by time series forecasting . We strongly recommend to develop a pan-European conservation strategy before the decline will reach its tipping point resulting into non-reversible loss of diversity in the European forest landscape.


Forests ◽  
2020 ◽  
Vol 11 (10) ◽  
pp. 1083
Author(s):  
Rajiv Chaudhary ◽  
Tilman Rönneburg ◽  
Matilda Stein Åslund ◽  
Karl Lundén ◽  
Mikael Brandström Durling ◽  
...  

Common ash (Fraxinus excelsior L.) is a tree species of significant ecological and economic importance that has suffered a devastating decline since the 1990s in Europe. Native ash species are being threatened by the alien invasive fungus Hymenoscyphus fraxineus, which causes ash dieback. The main goal of the study was to develop markers for traits related to tolerance to ash dieback and to investigate whether genotypes selected for tolerance were genetically different from susceptible wild populations. We phenotyped 326 ash trees from Sweden for disease severity and genotyped them using 63 amplicon-derived single-nucleotide polymorphism (SNP) markers derived from genes in 40 scaffolds spanning 8 MB in total, which represents approximately 1% of the ash genome. We used a mixed linear model to test for an association between genotypic variation at these loci and disease severity of ash. In total, two SNPs were found to have significant associations. One non-synonymous SNP associated with the disease severity of ash was found in a gene predicted to encode a subtilisin-related peptidase S8/S53 domain. A second marginally significant marker was associated with an LRR gene. Our results demonstrate an inexpensive time-effective method for generating genomic data that could have potential for use in future tree breeding programs and provide information for marker-assisted selection. Our study also showed a low differentiation between genotypes selected for disease tolerance and the wild population of ash representing a range of susceptibilities to ash dieback, indicating opportunities for further selection without significantly losing genetic diversity in the ash population.


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