Fungal endophytes in Fraxinus excelsior petioles and their in vitro antagonistic potential against the ash dieback pathogen Hymenoscyphus fraxineus

2022 ◽  
pp. 126961
Author(s):  
Piotr Bilański ◽  
Tadeusz Kowalski
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.


2020 ◽  
Vol 6 (4) ◽  
pp. 287
Author(s):  
Daniela Costa ◽  
Rui M. Tavares ◽  
Paula Baptista ◽  
Teresa Lino-Neto

An increase in cork oak diseases caused by Biscogniauxia mediterranea and Diplodia corticola has been reported in the last decade. Due to the high socio-economic and ecologic importance of this plant species in the Mediterranean Basin, the search for preventive or treatment measures to control these diseases is an urgent need. Fungal endophytes were recovered from cork oak trees with different disease severity levels, using culture-dependent methods. The results showed a higher number of potential pathogens than beneficial fungi such as cork oak endophytes, even in healthy plants. The antagonist potential of a selection of eight cork oak fungal endophytes was tested against B. mediterranea and D. corticola by dual-plate assays. The tested endophytes were more efficient in inhibiting D. corticola than B. mediterranea growth, but Simplicillium aogashimaense, Fimetariella rabenhorstii, Chaetomium sp. and Alternaria alternata revealed a high potential to inhibit the growth of both. Simplicillium aogashimaense caused macroscopic and microscopic mycelial/hyphal deformations and presented promising results in controlling both phytopathogens’ growth in vitro. The evaluation of the antagonistic potential of non-volatile and volatile compounds also revealed that A. alternata compounds could be further explored for inhibiting both pathogens. These findings provide valuable knowledge that can be further explored in in vivo assays to find a suitable biocontrol agent for these cork oak diseases.


2015 ◽  
Author(s):  
F. Muñoz ◽  
B. Marçais ◽  
J. Dufour ◽  
A. Dowkiw

AbstractSince the early 1990s, ash dieback due to the invasive ascomycete Hymenoscyphus fraxineus is threatening Fraxinus excelsior in most of its natural range. Previous studies reported significant levels of genetic variability for susceptibility in F. excelsior either in field or inoculation experiments. The present study was based on a field experiment planted in 1995, fifteen years before onset of the disease. Crown and collar status were monitored on 788 trees from 23 open-pollinated progenies originating from 3 French provenances. Susceptibility was modeled using a Bayesian approach where spatio-temporal effects were explicitly taken into account. Moderate narrow-sense heritability was found for Crown Dieback (CD, h2=0.42). This study is first to show that Collar Lesions are also heritable (h2=0.49 for prevalence and h2=0.42 for severity) and that there is significant genetic correlation (r=0.40) between the severities of both symptoms. There was no evidence for differences between Provenances. Family effects were detected, but computing Individual Breeding Values (IBV) showed that most of the genetic variation lies within families. In agreement with previous reports, early flushing correlates with better crown status. Consequences of these results in terms of management and breeding are discussed.


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

2016 ◽  
Vol 106 (12) ◽  
pp. 1535-1543 ◽  
Author(s):  
Facundo Muñoz ◽  
Benoît Marçais ◽  
Jean Dufour ◽  
Arnaud Dowkiw

Since the early 1990s, ash dieback due to the invasive ascomycete Hymenoscyphus fraxineus is threatening Fraxinus excelsior in most of its natural range. Previous studies reported significant levels of genetic variability in susceptibility in F. excelsior either in field or inoculation experiments. The present study was based on a field experiment planted in 1995, 15 years before onset of the disease. Crown and collar status were monitored on 777 trees from 23 open-pollinated progenies originating from three French provenances. Health status was modeled using a Bayesian approach where spatiotemporal effects were explicitly taken into account. Moderate narrow-sense heritability was found for crown dieback (h2 = 0.42). This study is first to show that resistance at the collar level is also heritable (h2 = 0.49 for collar lesions prevalence and h2 = 0.42 for their severity) and that there is significant genetic correlation (r = 0.40) between the severities of crown and collar symptoms. There was no evidence for differences between provenances. Family effects were detected, but computing individual breeding values showed that most of the genetic variation lies within families. In agreement with previous reports, early flushing correlates with healthier crown. Implications of these results in disease management and breeding are discussed.


Author(s):  
Stefan Klesse ◽  
Georg von Arx ◽  
Martin M Gossner ◽  
Christian Hug ◽  
Andreas Rigling ◽  
...  

Abstract Since the 1990s the invasive fungus Hymenoscyphus fraxineus has caused severe crown dieback and high mortality rates in Fraxinus excelsior in Europe. In addition to a strong genetic control of tolerance to the fungus, previous studies have found landscape heterogeneity to be an additional driver of variability in the severity of dieback symptoms. However, apart from climatic conditions related to heat and humidity influencing fungal infection success, the mechanistic understanding of why smaller or slower-growing trees are more susceptible to dieback remains less well understood. Here, we analyzed three stands in Switzerland with a unique setting of 8 years of data availability of intra-annual diameter growth and annual crown health assessments. We complemented this by ring width and quantitative wood anatomical measurements extending back before the monitoring started to investigate if wood anatomical adjustments can help better explain the size-related dieback phenomenon. We found that slower-growing trees or trees with smaller crowns already before the arrival of the fungus were more susceptible to dieback and mortality. Defoliation directly reduced growth as well as maximum earlywood vessel size, and the positive relationship between vessel size and growth rate caused a positive feedback amplifying and accelerating crown dieback. Measured non-structural carbohydrate (NSC) concentrations in the outermost five rings did not significantly vary between healthy and weakened trees, which translate into large differences in absolute available amount of NSCs. Thus, we hypothesize that a lack of NSCs (mainly sugars) leads to lower turgor pressure and smaller earlywood vessels in the following year. This might impede efficient water transport and photosynthesis, and be responsible for stronger symptoms of dieback and higher mortality rates in smaller and slower-growing trees.


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 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.


2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Shadi Eshghi Sahraei ◽  
Michelle Cleary ◽  
Jan Stenlid ◽  
Mikael Brandström Durling ◽  
Malin Elfstrand

Abstract Background With the expanding ash dieback epidemic that has spread across the European continent, an improved functional understanding of the disease development in afflicted hosts is needed. The study investigated whether differences in necrosis extension between common ash (Fraxinus excelsior) trees with different levels of susceptibility to the fungus Hymenoscyphus fraxineus are associated with, and can be explained by, the differences in gene expression patterns. We inoculated seemingly healthy branches of each of two resistant and susceptible ash genotypes with H. fraxineus grown in a common garden. Results Ten months after the inoculation, the length of necrosis on the resistant genotypes were shorter than on the susceptible genotypes. RNA sequencing of bark samples collected at the border of necrotic lesions and from healthy tissues distal to the lesion revealed relatively limited differences in gene expression patterns between susceptible and resistant genotypes. At the necrosis front, only 138 transcripts were differentially expressed between the genotype categories while 1082 were differentially expressed in distal, non-symptomatic tissues. Among these differentially expressed genes, several genes in the mevalonate (MVA) and iridoid pathways were found to be co-regulated, possibly indicating increased fluxes through these pathways in response to H. fraxineus. Comparison of transcriptional responses of symptomatic and non-symptomatic ash in a controlled greenhouse experiment revealed a relatively small set of genes that were differentially and concordantly expressed in both studies. This gene-set included the rate-limiting enzyme in the MVA pathway and a number of transcription factors. Furthermore, several of the concordantly expressed candidate genes show significant similarity to genes encoding players in the abscisic acid- or Jasmonate-signalling pathways. Conclusions A set of candidate genes, concordantly expressed between field and greenhouse experiments, was identified. The candidates are associated with hormone signalling and specialized metabolite biosynthesis pathways indicating the involvement of these pathways in the response of the host to infection by H. fraxineus.


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