Transmission of Fusarium boothii Mycovirus via Protoplast Fusion Causes Hypovirulence in Other Phytopathogenic Fungi

Lee, Kyung-Mi; Yu, Jisuk; Son, Moonil; Lee, Yin-Won; Kim, Kook‐Hyung

doi:10.1371/journal.pone.0021629

Cited by 74 publications

(72 citation statements)

References 42 publications

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“…Some hypovirulence-associated mycoviruses from B. cinerea and its related species Botrytis porri have been isolated and characterized (10,54,57), but these Botrytis viruses are unlikely to be used for control of diseases caused by these fungi under field conditions because they are easy to be eliminated via asexual propagation (see below). Expansion of the host range of mycoviruses has been accomplished for many fungus-mycovirus systems (7,8,24,(58)(59)(60)). …”

Section: Discussionmentioning

confidence: 99%

A Novel Partitivirus That Confers Hypovirulence on Plant Pathogenic Fungi

Xiao

Chéng

Tang

et al. 2014

J Virol

128

103

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Members of the family Partitiviridae have bisegmented double-stranded RNA (dsRNA) genomes and are not generally known to cause obvious symptoms in their natural hosts. An unusual partitivirus, Sclerotinia sclerotiorum partitivirus 1 (SsPV1/WF-1), conferred hypovirulence on its natural plant-pathogenic fungal host, Sclerotinia sclerotiorum strain WF-1. Cellular organelles, including mitochondria, were severely damaged. Hypovirulence and associated traits of strain WF-1 and SsPV1/WF-1 were readily cotransmitted horizontally via hyphal contact to different vegetative compatibility groups of S. sclerotiorum and interspecifically to Sclerotinia nivalis and Sclerotinia minor. S. sclerotiorum strain 1980 transfected with purified SsPV1/WF-1 virions also exhibited hypovirulence and associated traits similar to those of strain WF-1. Moreover, introduction of purified SsPV1/WF-1 virions into strain KY-1 of Botrytis cinerea also resulted in reductions in virulence and mycelial growth and, unexpectedly, enhanced conidial production. However, virus infection suppressed hyphal growth of most germinating conidia of B. cinerea and was eventually lethal to infected hyphae, since very few new colonies could develop following germ tube formation. Taken together, our results support the conclusion that SsPV1/WF-1 causes hypovirulence in Sclerotinia spp. and B. cinerea. Cryo-EM (cryo-electron microscopy) reconstruction of the SsPV1 particle shows that it has a distinct structure with similarity to the closely related partitiviruses Fusarium poae virus 1 and Penicillium stoloniferum virus F. These findings provide new insights into partitivirus biological activities and clues about molecular interactions between partitiviruses and their hosts. IMPORTANCEMembers of the Partitiviridae are believed to occur commonly in their phytopathogenic fungal and plant hosts. However, most partitiviruses examined so far appear to be associated with latent infections. Here we report a partitivirus, SsPV1/WF-1, that was isolated from a hypovirulent strain of Sclerotinia sclerotiorum and describe its biological and molecular features. We have demonstrated that SsPV1 confers hypovirulence. Furthermore, SsPV1 can infect and cause hypovirulence in Botrytis cinerea. Our study also suggests that SsPV1 has a vigorous ability to proliferate and spread via hyphal contact. SsPV1 can overcome vegetative incompatibility barriers and can be transmitted horizontally among different vegetative compatibility groups of S. sclerotiorum, even interspecifically. Cryo-EM reconstruction of SsPV1 shows that it has a distinct structure with similarity to closely related partitiviruses. Our studies exploit a novel system, SsPV1 and its hosts, which can provide the means to explore the mechanisms by which partitiviruses interact with their hosts.

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Section: Discussionmentioning

confidence: 99%

A Novel Partitivirus That Confers Hypovirulence on Plant Pathogenic Fungi

Xiao

Chéng

Tang

et al. 2014

J Virol

128

103

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“…In our study, the isolates growing at osmotic potential medium of -0.6 MPa and -1.8 MPa also showed a reduction of the mycelial growth. However, in the virus-infected Fusarium oxysporum strains when growing on PDA only slight morphological alterations were evident (Lee et al, 2011). Furthermore, it has been observed that Effect of mitovirus on mycelial growth of Gremmeniella abietina several Cryphonectria parasitica virus-infected strains grow as well as virus free isolates on most artificial media although they are incapable of producing grilling cankers on chestnut trees and sporulate poorly (McCabe et al 1999).…”

Section: Discussionmentioning

confidence: 99%

Effect of putative mitoviruses on in vitro growth of Gremmeniella abietina isolates under different laboratory conditions

2012

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Mitoviruses have been found in several forest pathogens (i.e. Cryphonectria parasitica, Gremmeniella abietina), and because they have been shown to reduce the virulence of host fungi there is a growing interest in studying their use as a biocontrol. This study was carried out to test the effect of temperature (5°C, 15°C, 25°C and 35°C), pH (4, 5, 7 and 9) and osmotic potential (-0.6, -1.2, -1.8 and -2.4 MPa) on the mycelial growth of seven G. abietina isolates under controlled laboratory conditions. Four of the isolates hosted mitoviruses and three of them did not. During the experiment, mycelial growth was recorded every week for a period of 8 weeks. Results showed no differences in growth behavior between mitovirus infected and non-infected isolates when placed under different pH modifications. However, the mitovirus-infected isolates presented larger mycelial growth than the mitovirus-free ones when at the fungi's optimal growing temperature of 15°C. When growing at certain osmotic potentials (-0.6 and -1.8 MPa) a reduction in growth of the mitovirus-infected isolates was observed. The results of this experiment suggest that mycelial growth among non-infected isolates and isolates naturally infected by mitovirus vary under different culture conditions, thus providing further insight into the effects of mitovirus on Gremmeniella abietina isolates.Key words: mitoviruses; Scleroderris canker; in vitro; biological control; Gremmeniella abietina; dsRNA. Resumen Efecto de posibles mitovirus en el crecimiento in vitro de aislados de Gremmeniella abietina bajo diferentes condiciones de laboratorioLos mitovirus son virus exclusivamente fúngicos que han sido aislados de algunos patógenos forestales (i.e. Cryphonectria parasitica, Gremmeniella abietina) y ya que pueden reducir la virulencia del hongo existe un creciente interés por su posible papel como agentes de control biológico. Se ha llevado a cabo un estudio para evaluar el efecto de la temperatura (5°C, 15°C, 25°C y 35°C), el pH (4, 5, 7 y 9) y el potencial osmótico (-0.6, -1.2, -1.8, -2.4 MPa) en el crecimiento micelial de siete aislados de G. abietina bajo condiciones controladas de laboratorio. Cuatro de los aislados albergaban mitovirus y tres de ellos no. Durante el experimento, el crecimiento micelial fue registrado semanalmente hasta completar 8 mediciones. Los aislados infectados con mitovirus presentaron mayor crecimiento micelial que los no infectados a la temperatura de crecimiento óptimo del hongo de 15°C. No se observaron efectos de la presencia de mitovirus entre los aislados infectados y los no infectados en los tratamientos de modificación del pH. Cuando se modificaron los potenciales osmóticos se observó una reducción del crecimiento micelial de los aislados infectados con mitovirus en los potenciales osmóticos de -0.6 y -1.8 MPa. Los resultados de este experimento sugieren que la presencia de los mitovirus afecta al crecimiento micelial del hongo bajo distintas condiciones de laboratorio. Este estudio proporciona un conocimiento más prof...

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“…All strains used in this study (Table 1) were stored in 25% (vol/vol) glycerol at Ϫ80°C and were reactivated on potato dextrose agar (PDA; Difco). For nucleic acid manipulation, all strains of F. graminearum were grown in 50 ml of a liquid complete medium (CM) (29) at 25°C with shaking (150 rpm) for 5 days, while strains of C. parasitica were grown in 50 ml of Endothia parasitica complete medium (29) at 26°C with shaking (120 rpm) for 5 days. Mycelia were harvested by filtration through miracloth (Calbiochem) and ground to a fine powder with liquid nitrogen in a mortar and pestle.…”

Section: Methodsmentioning

confidence: 99%

“…For extraction of genomic DNA, we used a previously described procedure (29). To construct a PCR fragment for deletion, overexpression, and complementation, a slightly modified double-joint (DJ) PCR strategy was applied for fusion of PCR products (20).…”

Section: Methodsmentioning

confidence: 99%

“…Protoplasts of the WT-VF strain (hereinafter, VF indicates virus free and VI indicates virus infected) were prepared by treating fresh mycelia grown in YPG liquid culture medium (0.3% yeast extract, 1% peptone, 2% glucose) for 3 h at 30°C with 1 M NH 4 Cl containing 10 mg/ml of Driselase (InterSpex Products) as previously described (29). Amplified PCR products were directly added, with 1 ml of polyethylene glycol (PEG) solution (60% PEG 3350, 10 mM Tris-HCl, pH 7.5, 10 mM CaCl 2 ), to protoplast suspensions as described above.…”

Section: Methodsmentioning

confidence: 99%

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The HEX1 Gene of Fusarium graminearum Is Required for Fungal Asexual Reproduction and Pathogenesis and for Efficient Viral RNA Accumulation of Fusarium graminearum Virus 1

Son

Lee

et al. 2013

J Virol

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bThe accumulation of viral RNA depends on many host cellular factors. The hexagonal peroxisome (Hex1) protein is a fungal protein that is highly expressed when the DK21 strain of Fusarium graminearum virus 1 (FgV1) infects its host, and Hex1 affects the accumulation of FgV1 RNA. The Hex1 protein is the major constituent of the Woronin body (WB), which is a peroxisome-derived electron-dense core organelle that seals the septal pore in response to hyphal wounding. To clarify the role of Hex1 and the WB in the relationship between FgV1 and Fusarium graminearum, we generated targeted gene deletion and overexpression mutants. Although neither HEX1 gene deletion nor overexpression substantially affected vegetative growth, both changes reduced the production of asexual spores and reduced virulence on wheat spikelets in the absence of FgV1 infection. However, the vegetative growth of deletion and overexpression mutants was increased and decreased, respectively, upon FgV1 infection compared to that of an FgV1-infected wild-type isolate. Viral RNA accumulation was significantly decreased in deletion mutants but was significantly increased in overexpression mutants compared to the viral RNA accumulation in the virus-infected wild-type control. Overall, these data indicate that the HEX1 gene plays a direct role in the asexual reproduction and virulence of F. graminearum and facilitates viral RNA accumulation in the FgV1-infected host fungus.T he interactions between viral elements and host factors are important for maintaining the infection cycles of RNA viruses in host cells. Viruses utilize numerous host factors that play essential roles in virus infection. Therefore, understanding the role(s) of host factors can provide insight into the molecular mechanism(s) of host and virus interactions. Relative to host factors affecting RNA viruses of animals and plants, host factors affecting fungal viruses (mycoviruses) are poorly understood.Several host and viral components required for virus life in cells have been identified and characterized in the model organism Saccharomyces cerevisiae. The knowledge obtained by studying S. cerevisiae as a host for several double-stranded RNA (dsRNA) and single-stranded RNA (ssRNA) viruses has greatly extended our understanding of mycovirus-host interaction (1). With respect to filamentous fungi, the host factors required for mycovirus replication and symptom induction have been well described for the interaction between the prototypic hypovirus Cryphonectria hypovirus 1 strain EP713 (CHV1) and its host, the chestnut blight fungus (Cryphonectria parasitica). One of these host factors, NAM-1, modulates symptom induction in the fungus in response to CHV1 infection (2). The hypovirus-responsive host transcription factor gene pro1 is required for female fertility of C. parasitica, development of its asexual spores, and the maintenance of CHV1 infection (3). The host gene Cpbir1 (bir1 of C. parasitica), which encodes the IAP (inhibitor of apoptosis protein) CpBir1 and which is required for fungal...

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Transmission of Fusarium boothii Mycovirus via Protoplast Fusion Causes Hypovirulence in Other Phytopathogenic Fungi

Cited by 74 publications

References 42 publications

A Novel Partitivirus That Confers Hypovirulence on Plant Pathogenic Fungi

A Novel Partitivirus That Confers Hypovirulence on Plant Pathogenic Fungi

Effect of putative mitoviruses on in vitro growth of Gremmeniella abietina isolates under different laboratory conditions

The HEX1 Gene of Fusarium graminearum Is Required for Fungal Asexual Reproduction and Pathogenesis and for Efficient Viral RNA Accumulation of Fusarium graminearum Virus 1

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