scholarly journals Modified small-scale batch procedure for isolation of dsRNA from Cryphonectria parasitica

2007 ◽  
Vol 88 (1) ◽  
pp. 27-29
Author(s):  
Jana Libantová ◽  
Jana Moravčíková ◽  
Katarína Adamčíková ◽  
Marek Kobza ◽  
Gabriela Juhásová
Keyword(s):  
Biological Control ◽  
Cost Savings ◽  
Cryphonectria Parasitica ◽  
Fungal Disease ◽  
Small Scale ◽  
Chestnut Blight ◽  
Chestnut Blight Fungus

Cytoplasmically-transmissible viral double-stranded RNAs of the genus Hypovirus cause reduced virulence (hypovirulence) in the chestnut blight fungus Cryphonectria parasitica. Biological control of this fungal disease is done by inoculating selected artificial hypovirulent strains of C. parasitica in the wounds of attacked chestnut trees and is followed by testing the transmissibility of dsRNA to C. parasitica isolates affecting these trees. Here we present a modified protocol of isolation and detection of dsRNA. The proposed procedure requires smaller amounts of fungal material for dsRNA detection and uses less reagents, thus resulting in appreciable cost savings.

scholarly journals Saprophytic Activity and Sporulation of Cryphonectria parasitica on Dead Chestnut Wood in Forests with Naturally Established Hypovirulence

2006 ◽  
Vol 96 (12) ◽  
pp. 1337-1344 ◽  
Author(s):  
S. Prospero ◽  
M. Conedera ◽  
U. Heiniger ◽  
D. Rigling
Keyword(s):  
Biological Control ◽  
Dead Wood ◽  
Transmission Rate ◽  
Virus Transmission ◽  
Cryphonectria Parasitica ◽  
Chestnut Blight ◽  
Chestnut Blight Fungus ◽  
The Mean ◽  
Chestnut Wood

Sustainable biological control of the chestnut blight fungus Crypho-nectria parasitica with hypovirulence depends on the production and dissemination of hypovirus-infected propagules of the pathogen. We investigated the ability of C. parasitica to sporulate and produce hypo-virus-infected spores on recently dead chestnut wood in coppice stands in southern Switzerland where hypovirulence has been naturally established. The number and type (active, inactive, or none) of cankers was assessed on experimentally cut and stacked stems, firewood stacks, and natural dead wood. Hypovirus-free and hypovirus-infected strains readily survived for more than 1 year in the chestnut blight cankers of the stacked stems. Sporulation of C. parasitica was observed on the surface of preexisting inactive and active cankers, as well as on newly colonized bark areas and was significantly more abundant than on comparable cankers on living stems. On all types of dead wood, we observed more stromata with perithecia than with pycnidia; however, a large proportion of the stromata was not differentiated. All perithecia examined yielded only hypovirus-free ascospores. The incidence of pycnidia that produced hypovirus-infected conidia ranged from 5% on natural dead wood to 41% on the experimental stacks. The mean virus transmission rate into conidia was 69%. Our study demonstrates a considerable saprophytic activity of C. parasitica on recently dead chestnut wood and supports the hypothesis of a role of this saprophytic phase in the epidemiology of hypovirulence.

scholarly journals 75 years of the chestnut blight fungus Cryphonectria parasitica (Murr.) Barr in Europe

2014 ◽  
pp. 82-85
Author(s):  
László Radócz ◽  
Gábor Görcsös ◽  
Gábor Tarcali ◽  
Gabriella Kovács ◽  
Qin Ling
Keyword(s):  
Biological Control ◽  
Cryphonectria Parasitica ◽  
Vegetative Compatibility ◽  
Chestnut Blight ◽  
Chestnut Blight Fungus ◽  
European Chestnut ◽  
High Prevalence ◽  
History Of ◽  
Short Time ◽  
High Diversity

The chestnut blight fungus Cryphonectria parasitica is a native pathogen in East Asia and has been introduced into North America and Europe. Historical records and population genetic studies revealed at least three major introduction events from Asia into Europe. Nowadays, chestnut blight is present in almost the entire distribution range of European chestnut, i.e. from the Iberian Peninsula to the Caucasus. The C. parasitica population in most countries has been studied in respect to the diversity of vegetative compatibility (vc) types and the occurrence of hypovirulence. The vc type diversity of the different populations varied considerably. Typically, a high diversity of vc types has been found in areas with a long history of chestnut blight and where sexual recombination between divergent genotypes commonly has occurred. On the other hand, newly established populations often showed a low diversity with only one, or a few vc types present. Hypovirulence, i.e. the occurrence of C. parasitica isolates infected by Cryphonectria hypovirus 1 has been found widespread in Europe. Natural dissemination and active biological control applications have lead to a high prevalence of the hypovirus and to the recovery of many chestnut stands. Virulent cankers became hypovirus-infected within a short time and ceased expansion. There is concern that the diversity of vegetative compatibility types could increase in Europe through sexual reproduction between C. parasitica genotypes originating from different introductions. A higher level of vegetative incompatibility would not only hamper hypovirus spread within a population but could also select for lower virulence in CHV-1 and subsequently lead to an erosion of biological control. Recent studies, however, indicate that the vc type barriers are not so restrictive than previously assumed and that so far no evidence for an erosion of biological control system in high diversity populations can be observed.

scholarly journals Biologische Bekämpfung des Kastanienrindenkrebses auf der Alpennordseite der Schweiz|Biological control of chestnut blight north of the Swiss Alps

2007 ◽  
Vol 158 (11) ◽  
pp. 342-348
Author(s):  
Ursula Heiniger
Keyword(s):  
Biological Control ◽  
Castanea Sativa ◽  
Cryphonectria Parasitica ◽  
Vegetative Compatibility ◽  
Swiss Alps ◽  
Chestnut Blight ◽  
Biologische Bekämpfung ◽  
Chestnut Blight Fungus ◽  
Low Rate

Since 1986 several isolated stands of chestnut (Castanea sativa) located north of the Swiss Alps have been infected with the chestnut blight fungus (Cryphonectria parasitica). At all sites (1–3), the diversity of the vegetative compatibility types was low. To control the disease, the hypovirus CHV1 was introduced at seventeen sites in five cantons. In total, 571 cankers were treated with local C. parasitica isolates containing CHV1. Re-inspection of the cankers one to two years after treatment demonstrated that the percentage of active cankers was significantly reduced in three cantons. Re-isolations of C. parasitica showed that the hypovirus persisted in 33% to 75% of the treated cankers and was disseminated to new cankers at a low rate. The difficulties and the potential of hypovirus treatment of small chestnut stands are discussed.

scholarly journals Role of Fresh Dead Wood in the Epidemiology and the Biological Control of the Chestnut Blight Fungus

Plant Disease ◽  
2019 ◽  
Vol 103 (3) ◽  
pp. 430-438 ◽  
Author(s):  
Joana Beatrice Meyer ◽  
Loïc Chalmandrier ◽  
Fabio Fässler ◽  
Christopher Schefer ◽  
Daniel Rigling ◽  
...  
Keyword(s):  
Biological Control ◽  
Dead Wood ◽  
Biocontrol Agent ◽  
Castanea Sativa ◽  
Cryphonectria Parasitica ◽  
Chestnut Blight ◽  
Chestnut Blight Fungus ◽  
Study Sites ◽  
Cryphonectria Hypovirus

The invasive fungus Cryphonectria parasitica, the causal agent of chestnut blight, is able to survive and sporulate on the bark of fresh dead Castanea sativa wood for at least 2 years. Here, we experimentally investigated the role of fresh dead wood in the epidemiology of chestnut blight, specifically in the spread of the hyperparasitic virus Cryphonectria hypovirus 1, which acts as biocontrol agent of C. parasitica. A total of 152 artificially initiated, virulent bark cankers in four chestnut stands were treated with virus-infected asexual spores originating either from sporulating dead wood or from a spore suspension. Molecular markers for both the virus and the fungal carrier were used to examine the spread of the applied biocontrol virus. Fourteen months after treatment, 42 to 76% of the conidial spray-treated cankers and 50 to 60% of the cankers exposed to a sporulating dead stem had been virus infected by the applied hypovirulent conidia in all four study sites. Virus infection reduced canker expansion and promoted canker healing (callusing). Thus, fresh chestnut dead wood may play an important role in supporting the successful spread of natural hypovirulence in chestnut forests. Further, combined with the application of virus-infected conidial suspensions, it may help promote the establishment of artificially released hypoviruses in chestnut stands to control chestnut blight.

scholarly journals Differential Transfer and Dissemination of Hypovirus and Nuclear and Mitochondrial Genomes of a Hypovirus-Infected Cryphonectria parasitica Strain after Introduction into a Natural Population

2003 ◽  
Vol 69 (7) ◽  
pp. 3767-3771 ◽  
Author(s):  
Patrik J. Hoegger ◽  
Ursula Heiniger ◽  
Ottmar Holdenrieder ◽  
Daniel Rigling
Keyword(s):  
Biological Control ◽  
Mitochondrial Dna ◽  
Nuclear Genome ◽  
Cryphonectria Parasitica ◽  
Plant Diseases ◽  
Mitochondrial Genomes ◽  
Chestnut Blight ◽  
Fungal Host ◽  
Chestnut Blight Fungus ◽  
Living Organisms

ABSTRACT Biological control of plant diseases generally requires release of living organisms into the environment. Cryphonectria hypoviruses function as biological control agents for the chestnut blight fungus, Cryphonectria parasitica, and hypovirus-infected C. parasitica strains can be used to treat infected trees. We used naturally occurring molecular marker polymorphisms to examine the persistence and dissemination of the three genomes of a hypovirus-infected C. parasitica strain, namely, the double-stranded RNA genome of Cryphonectria hypovirus 1 (CHV1) and the nuclear and mitochondrial genomes of its fungal host. The hypovirus-infected strain was experimentally introduced into a blight-infested chestnut coppice forest by treating 73 of 246 chestnut blight cankers. Two years after introduction, the hypovirus had disseminated to 36% of the untreated cankers and to 35% of the newly established cankers. Spread of the hypovirus was more frequent within treated sprout clusters than between sprout clusters. Mitochondrial DNA of the introduced fungus also was transferred into the resident C. parasitica population. Concomitant transfer of both the introduced hypovirus and mitochondrial DNA was detected in almost one-half of the treated cankers analyzed. The introduced mitochondrial DNA haplotype also was found in three resident isolates from newly established cankers. The nuclear genome of the introduced strain persisted in the treated cankers but did not spread beyond them.

scholarly journals Variation in Tolerance and Virulence in the Chestnut Blight Fungus-Hypovirus Interaction

2000 ◽  
Vol 66 (11) ◽  
pp. 4863-4869 ◽  
Author(s):  
Tobin L. Peever ◽  
Yir-Chung Liu ◽  
Paolo Cortesi ◽  
Michael G. Milgroom
Keyword(s):  
Biological Control ◽  
Genetic Variation ◽  
Local Population ◽  
Cryphonectria Parasitica ◽  
Conidial Germination ◽  
Relative Fitness ◽  
Chestnut Blight ◽  
Future Evolution ◽  
Chestnut Blight Fungus ◽  
Fungal Isolates

ABSTRACT Chestnut blight, caused by the fungus Cryphonectria parasitica, has been effectively controlled with double-stranded RNA hypoviruses in Europe for over 40 years. The marked reduction in the virulence of C. parasitica by hypoviruses is a phenomenon known as hypovirulence. This virus-fungus pathosystem has become a model system for the study of biological control of fungi with viruses. We studied variation in tolerance to hypoviruses in fungal hosts and variation in virulence among virus isolates from a local population in Italy. Tolerance is defined as the relative fitness of a fungal individual when infected with hypoviruses (compared to being uninfected); virulence is defined for each hypovirus as the reduction in fitness of fungal hosts relative to virus-free hosts. Six hypovirus-infected isolates of C. parasitica were sampled from the population, and each hypovirus was transferred into six hypovirus-free recipient isolates. The resulting 36 hypovirus-fungus combinations were used to estimate genetic variation in tolerance to hypoviruses, in hypovirus virulence, and in virus-fungus interactions. Four phenotypes were evaluated for each virus-fungus combination to estimate relative fitness: (i) sporulation, i.e., the number of asexual spores (conidia) produced; (ii) canker area on field-inoculated chestnut trees, (iii) vertical transmission of hypoviruses into conidia, and (iv) conidial germination. Two-way analysis of variance (ANOVA) revealed significant interactions (P < 0.001) between viruses and fungal isolates for sporulation and canker area but not for conidial germination or transmission. One-way ANOVA among hypoviruses (within each fungal isolate) and among fungal isolates (within each hypovirus) revealed significant genetic variation (P < 0.01) in hypovirus virulence and fungal tolerance within several fungal isolates, and hypoviruses, respectively. These interactions and the significant genetic variation in several fitness characters indicate the potential for future evolution of these characters. However, biological control is unlikely to break down due to evolution of tolerance to hypoviruses in the fungus because the magnitudes of tolerance and interactions were relatively small.

Crypt1, an active Ac-like transposon from the chestnut blight fungus, Cryphonectria parasitica

2001 ◽  
Vol 265 (4) ◽  
pp. 730-738 ◽  
Author(s):  
D. Linder-Basso ◽  
R. Foglia ◽  
P. Zhu ◽  
B.I. Hillman
Keyword(s):  
Cryphonectria Parasitica ◽  
Chestnut Blight ◽  
Chestnut Blight Fungus

scholarly journals The Morphology and the Biological Control of Cryphonectria parasitica

2012 ◽  
Vol 69 (1) ◽  
Author(s):  
Carmen Emilia PUIA ◽  
Daniela Andreea GRIGORESCU ◽  
Raluca Vasilica MICLEA
Keyword(s):  
Biological Control ◽  
Virulent Strain ◽  
Morphological Characteristics ◽  
Castanea Sativa ◽  
Cryphonectria Parasitica ◽  
Chestnut Blight ◽  
Dual Culture ◽  
Natural Form ◽  
Hypovirulent Strain ◽  
Endothia Parasitica

Cryphonectria parasitica  (Murr.) Bar [syn. Endothia parasitica (Murr. And.] (anamorf: Endothiella sp .) is the causal agent of chestnut bark disease or chestnut blight, disease which produced great damages throughout the world, for example, in Europe, the European chestnut tree ( Castanea sativa (P.) Mill) was heavily affected. Environmental concerns have focused attention on natural forms of disease control as an effective alternative to chemical pesticides. In the chestnut blight fungus, Cryphonectria parasitica deals with a natural form of biological control in which the virulence of a fungal pathogen is attenuated by an endogenous viral RNA genetic element- the hypovirulent strain. In our researches we picked samples of chestnut bark from different areas in Maramures county. We’ve isolated the fungus on PDA medium and we’ve studied the morphological characteristics of the usual virulent strain and we looked for a possible hypovirulent strain in order to study its capacity for biological control. The fungus develops in the bark and in cambium where forms a yellowish or brownish stroma and produces both conidia and ascospores. The pycnidia stromata break through the lenticels producing conidia and later in the same stroma develop the perithecia which produce ascospores. Both strains of the fungus were found in the research area. The hypovirulent strain had a slower development, showed no sporu lation and pigmentation “white cultural strain” and was tested in vitro for the capacity to convert the virulent isolates by dual culture tests.

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