scholarly journals Fungi Detected in the Previous Year’s Leaf Petioles of Fraxinus excelsior and Their Antagonistic Potential against Hymenoscyphus fraxineus

Forests ◽  
2021 ◽  
Vol 12 (10) ◽  
pp. 1412
Author(s):  
Tadeusz Kowalski ◽  
Piotr Bilański
Keyword(s):  
Its Region ◽  
Inhibition Zone ◽  
Fraxinus Excelsior ◽  
Rrna Gene ◽  
Dual Culture ◽  
Ash Dieback ◽  
Fusarium Lateritium ◽  
Hymenoscyphus Fraxineus ◽  
Dual Culture Assay

Studies on fungal communities in the previous year’s leaf petioles of Fraxinus excelsior found in litter in five ash stands in southern Poland were made in 2017. Fungi were identified on the basis of isolation from 300 surface sterilized leaf petioles and by in situ inventory of fruit bodies (on 600 petioles, in spring and autumn). Identification was based on morphology of colonies and fruit bodies, and sequencing of ITS region of the rRNA gene cluster. In total, 2832 isolates from 117 taxa (Ascomycota—100; Basidiomycota—15; Mucoromycota—2 taxa) were obtained with the isolation method. The most frequent taxa (with frequency >10%) were: Nemania serpens, Hymenoscyphus fraxineus, Alternaria sp. 1, Boeremia sp., Helotiales sp. 1, Epicoccum nigrum, Venturia fraxini, Fusarium sp., Fusarium lateritium, Nemania diffusa, Typhula sp. 2 (in descending order). In total, 45 taxa were detected with the in situ inventory method. Eleven taxa were classified as dominant: Hymenoscyphus fraxineus, Venturia fraxini, Leptosphaeria sp. 2, Cyathicula fraxinophila, Typhula sp. 2, Hypoderma rubi, Pyrenopeziza petiolaris, Cyathicula coronata, Hymenoscyphus scutula, Leptosphaeria sclerotioides and Hymenoscyphus caudatus. Among 202 leaf petioles colonized by H. fraxineus, 177 petioles also showed fructification of 26 other fungi. All the isolated saprotrophs were tested in dual-culture assay for antagonism to two strains of H. fraxineus. Three interaction types were observed: type A, mutual direct contact, when the two fungi meet along the contact line (occurred with 43.3% of test fungi); type B, with inhibition zone between colonies (with 46.9% of test fungi); type C, when the test fungus overgrows the colony of H. fraxineus (with 9.8% of test fungi). The possible contribution of the fungal saprotrophs in limiting of the expansion of H. fraxineus in ash leaf petioles, which may result in reduction in the inoculum of ash dieback causal agent, is discussed.

scholarly journals Rising out of the ashes: additive genetic variation for susceptibility to Hymenoscyphus fraxineus in Fraxinus excelsior

2015 ◽  
Author(s):  
F. Muñoz ◽  
B. Marçais ◽  
J. Dufour ◽  
A. Dowkiw
Keyword(s):  
Genetic Variation ◽  
Fraxinus Excelsior ◽  
Narrow Sense ◽  
Narrow Sense Heritability ◽  
Ash Dieback ◽  
Family Effects ◽  
Hymenoscyphus Fraxineus ◽  
Natural Range ◽  
Spatio Temporal ◽  
Crown Dieback

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.

scholarly journals Molecular Characterization of Trichoderma spp. Isolates by Internal Transcribed Spacer (ITS) Region Sequencing Technique and its Use as a Biocontrol Agent

2020 ◽  
Vol 14 (1) ◽  
pp. 70-77
Author(s):  
Ziyaul Haque ◽  
Mohammed S. Iqbal ◽  
Ausaf Ahmad ◽  
Mohd S. Khan ◽  
Jyoti Prakash
Keyword(s):  
Internal Transcribed Spacer ◽  
Biocontrol Agent ◽  
Its Region ◽  
Antagonistic Activity ◽  
Rrna Gene ◽  
Dual Culture ◽  
Sequencing Analysis ◽  
Trichoderma Spp ◽  
Sequencing Technique ◽  
Its Region Sequencing

Objective: In the present investigation, Trichoderma spp., isolated from rhizospheric soil, has been identified by Internal Transcribed Spacer (ITS) region sequencing technique and its antagonistic activity was evaluated against A. niger. Methods: The sequencing analysis was done with its ITS1 region of the rRNA gene. Using the ITS1 amplified products for all isolated fungi, a bi-directional DNA sequencing was done with high quality bases (>98% - 100%). Antagonistic activity was done using dual culture technique. Results: All of the ITS1 nucleotide sequences obtained in this study matched 97% - 100% with the published sequence of Trichoderma spp. The results confirmed the strains as T. asperellum and T. viride with gene bank accession no. (ZTa); MK937669 and (ZTv); MK503705, respectively. When phylogenetic analysis was done for the isolates, the optimal tree with the sum of branch length = 0.69585023 and 0.10077756 for T. asperellum and T. viride, respectively, was observed. There were a total of 678 and 767 for T. asperellum and T. viride positions in the final dataset, respectively. Antagonistic activity was done for the isolated strains of Trichoderma spp. against A. niger, and it was found that T. asperellum showed maximum antagonistic activity (79.33±7.09%). Conclusion: The findings prolong the genome availability for relative investigations pointing out phenotypic variances to compare with Trichoderma genetic diversity. The present investigation delivered the Bases of future studies for better knowledge in understanding the complicated connections of Trichoderma spp. to be used as an effective biocontrol agent.

scholarly journals Estimating mortality rates of European ash ( Fraxinus excelsior ) under the ash dieback ( Hymenoscyphus fraxineus ) epidemic

2018 ◽  
Vol 1 (1) ◽  
pp. 48-58 ◽  
Author(s):  
Tim L. R. Coker ◽  
Jiří Rozsypálek ◽  
Anne Edwards ◽  
Tony P. Harwood ◽  
Louise Butfoy ◽  
...  
Keyword(s):  
Mortality Rates ◽  
Fraxinus Excelsior ◽  
Ash Dieback ◽  
Hymenoscyphus Fraxineus ◽  
European Ash

scholarly journals Rising Out of the Ashes: Additive Genetic Variation for Crown and Collar Resistance to Hymenoscyphus fraxineus in Fraxinus excelsior

2016 ◽  
Vol 106 (12) ◽  
pp. 1535-1543 ◽  
Author(s):  
Facundo Muñoz ◽  
Benoît Marçais ◽  
Jean Dufour ◽  
Arnaud Dowkiw
Keyword(s):  
Genetic Variation ◽  
Health Status ◽  
Fraxinus Excelsior ◽  
Narrow Sense ◽  
Narrow Sense Heritability ◽  
Ash Dieback ◽  
Family Effects ◽  
Hymenoscyphus Fraxineus ◽  
Natural Range ◽  
Crown Dieback

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.

scholarly journals Amplifying feedback loop between growth and wood anatomical characteristics of Fraxinus excelsior explains size-related susceptibility to ash dieback

2020 ◽  
Author(s):  
Stefan Klesse ◽  
Georg von Arx ◽  
Martin M Gossner ◽  
Christian Hug ◽  
Andreas Rigling ◽  
...  
Keyword(s):  
Landscape Heterogeneity ◽  
Turgor Pressure ◽  
Ring Width ◽  
Mortality Rates ◽  
Data Availability ◽  
Fraxinus Excelsior ◽  
Ash Dieback ◽  
Vessel Size ◽  
Hymenoscyphus Fraxineus ◽  
Crown Dieback

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.

scholarly journals Diplodia fraxini and Diplodia subglobosa: The Main Species Associated with Cankers and Dieback of Fraxinus excelsior in North-Eastern Italy

Forests ◽  
2020 ◽  
Vol 11 (8) ◽  
pp. 883
Author(s):  
Benedetto T. Linaldeddu ◽  
Francesco Bottecchia ◽  
Carlo Bregant ◽  
Lucia Maddau ◽  
Lucio Montecchio
Keyword(s):  
Dna Sequences ◽  
Fungal Pathogens ◽  
Fungal Species ◽  
Fraxinus Excelsior ◽  
Limited Information ◽  
Ash Dieback ◽  
Alpine Regions ◽  
Hymenoscyphus Fraxineus ◽  
North Eastern ◽  
Friuli Venezia Giulia

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.

scholarly journals Isolation of actinomycetes from maize rhizosphere from Kupang, East Nusa Tenggara Province, and evaluation of their antibacterial, antifungal, and extracellular enzyme activity

2018 ◽  
Vol 23 (1) ◽  
pp. 40
Author(s):  
Umi Fatmawati ◽  
Yulin Lestari ◽  
Anja Meryandini ◽  
Abdjad Asih Nawangsih ◽  
Aris Tri Wahyudi
Keyword(s):  
Extracellular Enzymes ◽  
Sclerotium Rolfsii ◽  
Extracellular Enzyme ◽  
Rrna Gene ◽  
Dual Culture ◽  
Lytic Enzymes ◽  
Genus Streptomyces ◽  
Maize Rhizosphere ◽  
Dual Culture Assay ◽  
The One

Actinomycetes are the one of the components of the rhizospheric microbial population and useful for producing secondary metabolites such as lytic enzymes, antibiotics, and antifungal. The aim of the study was to isolate the actinomycetes from maize rhizosphere collected from Kupang, East Nusa Tenggara. The screening was focused on the actinomycetes that showed the ability to produce antibacterial, antifungal, and extracellular enzymes such as amylase, cellulase, and protease. The actinomycetes were isolated using Humic-Acid Vitamin B (HV) agar media. The antagonistic assay was tested against Escherichia coli, Staphylococcus aureus, Sclerotium rolfsii and Fusarium oxysporum. Isolate JKP-8 was an isolate that showed the highest activity in inhibiting the growth of E. coli and S. aureus bacteria. Isolate JKP-5 showed the highest activity in inhibiting the growth of F.oxysporum. There were no actinomycetes isolates that showed an ability to inhibit the growth of S. rolfsii fungus based on dual culture assay. JKP-3 and JKP-4 isolates exhibited the highest ability to hydrolyze amylum, while JKP-5 and JKP-8 isolates exhibited the highest ability to hydrolyze CMC. The results of the amplification of 16S rRNA gene in selected potential isolates JKP 5 and JKP 8 indicated that both isolates belong to the genus Streptomyces.

scholarly journals Decline of Fraxinus excelsior L. in parks of Saint Petersburg: Who is to blame – Hymenoscyphus fraxineus or Diplodia spp.?

2020 ◽  
Vol 73 (1) ◽  
pp. 43-51
Author(s):  
Dmitrii A. Shabunin ◽  
Andrey V. Selikhovkin ◽  
Elena Yu. Varentsova ◽  
Dmitry L. Musolin
Keyword(s):  
Light Microscopy ◽  
Fraxinus Excelsior ◽  
Ash Dieback ◽  
Saint Petersburg ◽  
Hymenoscyphus Fraxineus ◽  
Ash Trees ◽  
Fallen Leaves ◽  
European Ash

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.

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