RESUMEN
In this study, a novel mitovirus, tentatively designated as "Alternaria alternata mitovirus 2" (AaMV2), was isolated from the fungus Alternaria alternata f. sp. mali causing apple leaf blotch disease. The complete genome of AaMV2 is 3,157 nucleotides in length, with an A+U content of 68.10%. The genome has a single large open reading frame (ORF) encoding an RNA-dependent RNA polymerase (RdRp) protein with a molecular mass of 98.10 kDa. BLAST analysis revealed that AaMV2 has the highest sequence identity to Leptosphaeria biglobosa mitovirus 6, with 79.76% and 82.86% identity at the amino acid and nucleotide level, respectively. Phylogenetic analysis suggested that AaMV2 is a new member of the genus Duamitovirus within the family Mitoviridae. This is the first report of the complete genome sequence analysis of a mitovirus in A. alternata.
Asunto(s)
Alternaria , Virus Fúngicos , Genoma Viral , Malus , Sistemas de Lectura Abierta , Filogenia , Enfermedades de las Plantas , Virus ARN , Secuenciación Completa del Genoma , Alternaria/virología , Alternaria/genética , Enfermedades de las Plantas/microbiología , Malus/microbiología , Malus/virología , Virus Fúngicos/genética , Virus Fúngicos/aislamiento & purificación , Virus Fúngicos/clasificación , Virus ARN/genética , Virus ARN/aislamiento & purificación , Proteínas Virales/genética , ARN Viral/genética , ARN Polimerasa Dependiente del ARN/genética , Composición de Base , Hojas de la Planta/microbiología , Hojas de la Planta/virología , Secuencia de BasesRESUMEN
Here, we characterized a novel mitovirus from the fungus Nigrospora oryzae, which was named "Nigrospora oryzae mitovirus 3" (NoMV3). The NoMV3 genome is 2,492 nt in length with a G + C content of 33%, containing a single large open reading frame (ORF) using the fungal mitochondrial genetic code. The ORF encodes an RNA-dependent RNA polymerase (RdRp) of 775 amino acids with a molecular mass of 88.75 kDa. BLASTp analysis revealed that the RdRp of NoMV3 had 68.6%, 50.6%, and 48.6% sequence identity to those of Nigrospora oryzae mitovirus 2, Suillus luteus mitovirus 6, and Fusarium proliferatum mitovirus 3, respectively, which belong to the genus Unuamitovirus within the family Mitoviridae. Phylogenetic analysis based on amino acid sequences supported the classification of NoMV3 as a member of a new species in the genus Unuamitovirus within the family Mitoviridae.
Asunto(s)
Ascomicetos , Virus Fúngicos , Genoma Viral , Sistemas de Lectura Abierta , Filogenia , Enfermedades de las Plantas , Virus ARN , ARN Polimerasa Dependiente del ARN , Genoma Viral/genética , Virus Fúngicos/genética , Virus Fúngicos/clasificación , Virus Fúngicos/aislamiento & purificación , Virus ARN/genética , Virus ARN/clasificación , Virus ARN/aislamiento & purificación , ARN Polimerasa Dependiente del ARN/genética , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/virología , Ascomicetos/virología , Ascomicetos/genética , ARN Viral/genética , Proteínas Virales/genética , Composición de Base , Secuencia de AminoácidosRESUMEN
Monilinia fructicola is one of the most devastating fungal diseases of rosaceous fruit crops, both in the field and postharvest, causing significant yield losses. Here, we report the discovery of a novel positive single-stranded RNA virus, Monilinia fructicola hypovirus 3 (MfHV3), in a strain (hf-1) of the phytopathogenic fungus Monilinia fructicola. The complete genome of MfHV3 is 9259 nucleotides (nt) in length and contains a single large open reading frame (ORF) from nt position 462 to 8411. This ORF encodes a polyprotein with three conserved domains, namely UDP-glycosyltransferase, RNA-dependent RNA polymerase (RdRp), and DEAD-like helicase. The MfHV3 polyprotein shares the highest similarity with Colletotrichum camelliae hypovirus 1. Phylogenetic analysis indicated that MfHV3 clustered with members of the genus Betahypovirus within the family Hypoviridae. Taken together, the results of genomic organization comparisons, amino acid sequence alignments, and phylogenetic analysis convincingly show that MfHV3 is a new member of the genus Betahypovirus, family Hypoviridae.
Asunto(s)
Ascomicetos , Virus Fúngicos , Genoma Viral , Sistemas de Lectura Abierta , Filogenia , Enfermedades de las Plantas , Ascomicetos/virología , Ascomicetos/genética , Virus Fúngicos/genética , Virus Fúngicos/clasificación , Virus Fúngicos/aislamiento & purificación , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/virología , ARN Viral/genética , Proteínas Virales/genética , Secuenciación Completa del Genoma , Virus ARN/genética , Virus ARN/clasificación , Virus ARN/aislamiento & purificación , ARN Polimerasa Dependiente del ARN/genética , Secuencia de AminoácidosRESUMEN
Here, we report a novel ourmia-like mycovirus, named "Phomopsis asparagi magoulivirus 1" (PaMV1), derived from the phytopathogenic fungus Phomopsis asparagi. The genome of PaMV1 consists of a positive-sense single-stranded RNA (+ ssRNA) that is 2,639 nucleotides in length, with a GC content of 57.13%. It contains a single open reading frame (ORF) encoding a putative RNA-dependent RNA polymerase (RdRp) consisting of 686 amino acids with a molecular mass of 78.57 kDa. Phylogenetic analysis based on RdRp sequences revealed that PaMV1 grouped together with Diaporthe gulyae magoulivirus 1 (DgMV1) in a distinct clade. Sequence comparisons and phylogenetic analysis suggest that PaMV1 is a novel member of the genus Magoulivirus, family Botourmiaviridae.
Asunto(s)
Virus Fúngicos , Genoma Viral , Sistemas de Lectura Abierta , Phomopsis , Filogenia , ARN Viral , Virus Fúngicos/genética , Virus Fúngicos/clasificación , Virus Fúngicos/aislamiento & purificación , Phomopsis/virología , ARN Viral/genética , Secuenciación Completa del Genoma , ARN Polimerasa Dependiente del ARN/genética , Composición de Base , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/virología , Proteínas Virales/genética , Secuencia de Bases , Virus ARN/genética , Virus ARN/aislamiento & purificación , Virus ARN/clasificaciónRESUMEN
The virus family Phenuiviridae (order Hareavirales, comprising segmented negative-sense single stranded RNA viruses) has highly diverse members that are known to infect animals, plants, protozoans, and fungi. In this study, we identified a novel phenuivirus infecting a strain of the entomopathogenic fungus Cordyceps javanica isolated from a small brown plant hopper (Laodelphax striatellus), and this virus was tentatively named "Cordyceps javanica negative-strand RNA virus 1" (CjNRSV1). The CjNRSV1 genome consists of three negative-sense single stranded RNA segments (RNA1-3) with lengths of 7252, 2401, and 1117 nt, respectively. The 3'- and 5'-terminal regions of the RNA1, 2, and 3 segments have identical sequences, and the termini of the RNA segments are complementary to each other, reflecting a common characteristic of viruses in the order Hareavirales. RNA1 encodes a large protein (â¼274 kDa) containing a conserved domain for the bunyavirus RNA-dependent RNA polymerase (RdRP) superfamily, with 57-80% identity to the RdRP encoded by phenuiviruses in the genus Laulavirus. RNA2 encodes a protein (â¼79 kDa) showing sequence similarity (47-63% identity) to the movement protein (MP, a plant viral cell-to-cell movement protein)-like protein (MP-L) encoded by RNA2 of laulaviruses. RNA3 encodes a protein (â¼28 kDa) with a conserved domain of the phenuivirid nucleocapsid protein superfamily. Phylogenetic analysis using the RdRPs of various phenuiviruses and other unclassified phenuiviruses showed CjNRSV1 to be grouped with established members of the genus Laulavirus. Our results suggest that CjNRSV1 is a novel fungus-infecting member of the genus Laulavirus in the family Phenuiviridae.
Asunto(s)
Cordyceps , Genoma Viral , Filogenia , ARN Viral , Cordyceps/genética , ARN Viral/genética , Virus Fúngicos/clasificación , Virus Fúngicos/genética , Virus Fúngicos/aislamiento & purificación , Proteínas Virales/genética , Virus ARN de Sentido Negativo/genética , Virus ARN de Sentido Negativo/clasificación , ARN Polimerasa Dependiente del ARN/genética , Virus ARN/genética , Virus ARN/clasificación , Virus ARN/aislamiento & purificación , Secuencia de Aminoácidos , Sistemas de Lectura AbiertaRESUMEN
In this study, we identified a novel partitivirus, named "Cordyceps militaris partitivirus 1" (CmPV1), in Cordyceps militaris strain RCEF7506. The complete genome of CmPV1 comprises two segments, dsRNA1 and dsRNA2, each encoding a single protein. dsRNA1 (2,206 bp) encodes an RNA-dependent RNA polymerase (RdRp), and dsRNA2 (2,256 bp) encodes a coat protein (CP). Sequence analysis revealed that dsRNA1 has the highest similarity to that of Bipolaris maydis partitivirus 2 (BmPV2), whereas dsRNA2 shows the highest similarity to human blood-associated partitivirus (HuBPV). Phylogenetic analysis based on RdRp sequences suggests that CmPV1 is a new member of the genus Betapartitivirus of the family Partitiviridae. This is the first documentation of a betapartitivirus infecting the entomopathogenic fungus C. militaris.
Asunto(s)
Cordyceps , Virus Fúngicos , Genoma Viral , Filogenia , Virus ARN , Cordyceps/genética , Cordyceps/virología , Cordyceps/aislamiento & purificación , Genoma Viral/genética , Virus Fúngicos/genética , Virus Fúngicos/aislamiento & purificación , Virus Fúngicos/clasificación , Virus ARN/genética , Virus ARN/aislamiento & purificación , Virus ARN/clasificación , ARN Viral/genética , ARN Polimerasa Dependiente del ARN/genética , Sistemas de Lectura Abierta , Proteínas Virales/genética , Proteínas de la Cápside/genéticaRESUMEN
Rice sheath blight, caused by the soil-borne fungus Rhizoctonia solani (teleomorph: Thanatephorus cucumeris, Basidiomycota), is one of the most devastating phytopathogenic fungal diseases and causes yield loss. Here, we report on a very high prevalence (100%) of potential virus-associated double-stranded RNA (dsRNA) elements for a collection of 39 fungal strains of R. solani from the rice sheath blight samples from at least four major rice-growing areas in the Philippines and a reference isolate from the International Rice Research Institute, showing different colony phenotypes. Their dsRNA profiles suggested the presence of multiple viral infections among these Philippine R. solani populations. Using next-generation sequencing, the viral sequences of the three representative R. solani strains (Ilo-Rs-6, Tar-Rs-3, and Tar-Rs-5) from different rice-growing areas revealed the presence of at least 36 viruses or virus-like agents, with the Tar-Rs-3 strain harboring the largest number of viruses (at least 20 in total). These mycoviruses or their candidates are believed to have single-stranded RNA or dsRNA genomes and they belong to or are associated with the orders Martellivirales, Hepelivirales, Durnavirales, Cryppavirales, Ourlivirales, and Ghabrivirales based on their coding-complete RNA-dependent RNA polymerase sequences. The complete genome sequences of two novel RNA viruses belonging to the proposed family Phlegiviridae and family Mitoviridae were determined.
Asunto(s)
Oryza , Filogenia , Enfermedades de las Plantas , Virus ARN , Rhizoctonia , Rhizoctonia/virología , Rhizoctonia/genética , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/virología , Oryza/microbiología , Oryza/virología , Virus ARN/genética , Virus ARN/aislamiento & purificación , Virus ARN/clasificación , Genoma Viral , ARN Viral/genética , Secuenciación de Nucleótidos de Alto Rendimiento , ARN Bicatenario/genética , Virus Fúngicos/genética , Virus Fúngicos/clasificación , Virus Fúngicos/aislamiento & purificación , Filipinas , TranscriptomaRESUMEN
Peanut stem rot is a soil-borne disease caused by Agroathelia rolfsii. It occurs widely and seriously affects the peanut yield in most peanut-producing areas. The mycoviruses that induce the hypovirulence of some plant pathogenic fungi are potential resources for the biological control of fungal diseases in plants. Thus far, few mycoviruses have been found in A. rolfsii. In this study, two mitoviruses, namely, Agroathelia rolfsii mitovirus 1 (ArMV1) and Agroathelia rolfsii mitovirus 2 (ArMV2), were identified from the weakly virulent A. rolfsii strain GP3-1, and they were also found in other A. rolfsii isolates. High amounts of ArMV1 and ArMV2in the mycelium could reduce the virulence of A. rolfsii strains. This is the first report on the existence of mitoviruses in A. rolfsii. The results of this study may provide insights into the classification and evolution of mitoviruses in A. rolfsii and enable the exploration of the use of mycoviruses as biocontrol agents for the control of peanut stem rot.
Asunto(s)
Arachis , Virus Fúngicos , Filogenia , Enfermedades de las Plantas , Virus ARN , Arachis/virología , Arachis/microbiología , Enfermedades de las Plantas/virología , Enfermedades de las Plantas/microbiología , Virus ARN/genética , Virus ARN/clasificación , Virus ARN/aislamiento & purificación , Virus Fúngicos/clasificación , Virus Fúngicos/aislamiento & purificación , Virus Fúngicos/genética , Genoma Viral , Virulencia , ARN Viral/genéticaRESUMEN
Mycoviruses are usually transmitted horizontally via hyphal anastomosis and vertically through sporulation in natural settings. Oyster mushroom spherical virus (OMSV) is a mycovirus that infects Pleurotus ostreatus, with horizontal transmission via hyphal anastomosis. However, whether OMSV can be vertically transmitted is unclear. This study aimed to investigate the transmission characteristics of OMSV to progeny via basidiospores and horizontally to a new host. A total of 37 single-basidiospore offspring were obtained from OMSV-infected P. ostreatus and Pleurotus pulmonarius for Western blot detection of OMSV. The OMSV-carrying rate among monokaryotic isolates was 19% in P. ostreatus and 44% in P. pulmonarius. Then, OMSV-free and OMSV-infected monokaryotic isolates were selected for hybridization with harvested dikaryotic progeny strains. Western blot analyses of the offspring revealed that the OMSV transmission efficiency was 50% in P. ostreatus and 75% in P. pulmonarius, indicating vertical transmission via sexual basidiospores. Furthermore, we observed the horizontal transfer of OMSV from P. pulmonarius to Pleurotus floridanus. OMSV infection in P. floridanus resulted in significant inhibition of mycelial growth and yield loss. This study was novel in reporting the vertical transmission of OMSV through basidiospores, and its infection and pathogenicity in a new host P. floridanus.
Asunto(s)
Virus Fúngicos , Pleurotus , Esporas Fúngicas , Pleurotus/virología , Esporas Fúngicas/crecimiento & desarrollo , Virus Fúngicos/fisiologíaRESUMEN
The genus Alternaria comprises many important fungal pathogens that infect a wide variety of organisms. In this report, we present the discovery of a new double-stranded RNA (dsRNA) mycovirus called Alternaria botybirnavirus 2 (ABRV2) from a phytopathogenic strain, XC21-21C, of Alternaria sp. isolated from diseased tobacco leaves in China. The ABRV2 genome consists of two dsRNA components, namely dsRNA1 and dsRNA2, with lengths of 6,162 and 5,865 base pairs (bp), respectively. Each of these genomic dsRNAs is monocistronic, encoding hypothetical proteins of 201.6 kDa (P1) and 2193.3 kDa (P2). ABRV2 P1 and P2 share 50.54% and 63.13% amino acid sequence identity with the corresponding proteins encoded by dsRNA1 of Alternaria botybirnavirus 1 (ABRV1). Analysis of its genome organization and phylogenetic analysis revealed that ABRV2 is a new member of the genus Botybirnavirus.
Asunto(s)
Alternaria , Virus Fúngicos , Genoma Viral , Nicotiana , Filogenia , Enfermedades de las Plantas , ARN Bicatenario , ARN Viral , Alternaria/virología , Alternaria/genética , Nicotiana/virología , Nicotiana/microbiología , Virus Fúngicos/genética , Virus Fúngicos/clasificación , Virus Fúngicos/aislamiento & purificación , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/virología , ARN Viral/genética , ARN Bicatenario/genética , China , Virus ARN Bicatenario/genética , Virus ARN Bicatenario/aislamiento & purificación , Virus ARN Bicatenario/clasificación , Hojas de la Planta/virología , Hojas de la Planta/microbiología , Proteínas Virales/genéticaRESUMEN
Non-self recognition is a fundamental aspect of life, serving as a crucial mechanism for mitigating proliferation of molecular parasites within fungal populations. However, studies investigating the potential interference of plants with fungal non-self recognition mechanisms are limited. Here, we demonstrate a pronounced increase in the efficiency of horizontal mycovirus transmission between vegetatively incompatible Sclerotinia sclerotiorum strains in planta as compared to in vitro. This increased efficiency is associated with elevated proline concentration in plants following S. sclerotiorum infection. This surge in proline levels attenuates the non-self recognition reaction among fungi by inhibition of cell death, thereby facilitating mycovirus transmission. Furthermore, our field experiments reveal that the combined deployment of hypovirulent S. sclerotiorum strains harboring hypovirulence-associated mycoviruses (HAVs) together with exogenous proline confers substantial protection to oilseed rape plants against virulent S. sclerotiorum. This unprecedented discovery illuminates a novel pathway by which plants can counteract S. sclerotiorum infection, leveraging the weakening of fungal non-self recognition and promotion of HAVs spread. These promising insights provide an avenue to explore for developing innovative biological control strategies aimed at mitigating fungal diseases in plants by enhancing the efficacy of horizontal HAV transmission.
Asunto(s)
Ascomicetos , Virus Fúngicos , Enfermedades de las Plantas , Prolina , Virus Fúngicos/fisiología , Virus Fúngicos/genética , Prolina/metabolismo , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/virología , Ascomicetos/virología , Ascomicetos/fisiología , Brassica napus/microbiología , Brassica napus/virología , Virulencia , Interacciones Huésped-PatógenoRESUMEN
Although most known viruses infecting fungi pathogenic to higher eukaryotes are asymptomatic or reduce the virulence of their host fungi, those that confer hypervirulence to entomopathogenic fungus still need to be explored. Here, we identified and studied a novel mycovirus in Metarhizium flavoviride, isolated from small brown planthopper (Laodelphax striatellus). Based on molecular analysis, we tentatively designated the mycovirus as Metarhizium flavoviride partitivirus 1 (MfPV1), a species in genus Gammapartitivirus, family Partitiviridae. MfPV1 has two double-stranded RNAs as its genome, 1,775 and 1,575 bp in size respectively, encapsidated in isometric particles. When we transfected commercial strains of Metarhizium anisopliae and Metarhizium pingshaense with MfPV1, conidiation was significantly enhanced (t test; P-value < 0. 01), and the significantly higher mortality rates of the larvae of diamondback moth (Plutella xylostella) and fall armyworm (Spodoptera frugiperda), two important lepidopteran pests were found in virus-transfected strains (ANOVA; P-value < 0.05). Transcriptomic analysis showed that transcript levels of pathogenesis-related genes in MfPV1-infected M. anisopliae were obviously altered, suggesting increased production of metarhizium adhesin-like protein, hydrolyzed protein, and destruxin synthetase. Further studies are required to elucidate the mechanism whereby MfPV1 enhances the expression of pathogenesis-related genes and virulence of Metarhizium to lepidopteran pests. This study presents experimental evidence that the transfection of other entomopathogenic fungal species with a mycovirus can confer significant hypervirulence and provides a good example that mycoviruses could be used as a synergistic agent to enhance the biocontrol activity of entomopathogenic fungi.
Asunto(s)
Virus Fúngicos , Metarhizium , Metarhizium/patogenicidad , Metarhizium/genética , Animales , Virulencia/genética , Virus Fúngicos/genética , Control Biológico de Vectores/métodos , Mariposas Nocturnas/microbiología , Mariposas Nocturnas/virología , Genoma Viral , FilogeniaRESUMEN
The extensive use of high-throughput sequencing (HTS) has significantly advanced and transformed our comprehension of virus diversity, especially in intricate settings like soil and biological specimens. In this study, we delved into mycovirus sequence surveys within mycorrhizal fungus species Terfezia claveryi, through employing HTS with total double-stranded RNA (dsRNA) extracts. Our findings revealed the presence of four distinct members from the Alsuviricetes class, one flexivirus designated as Terfezia claveryi flexivirus 1 (TcFV1) and three endornaviruses (TcEV1, TcEV2, and TcEV3) in two different T. claveryi isolates. TcFV1, a member of the order Tymovirales, exhibits a unique genome structure and sequence features. Through in-depth analyses, we found that it shares sequence similarities with other deltaflexiviruses and challenges existing Deltaflexiviridae classification. The discovery of TcFV1 adds to the genomic plasticity of mycoviruses within the Tymovirales order, shedding light on their evolutionary adaptations. Additionally, the three newly discovered endornaviruses (TcEV1, TcEV2, and TcEV3) in T. claveryi exhibited limited sequence similarities with other endornaviruses and distinctive features, including conserved domains like DEAD-like helicase, ATPases Associated with Diverse Cellular Activities (AAA ATPase), and RNA dependent RNA polymerase (RdRp), indicating their classification as members of new species within the Alphaendornavirus genus. In conclusion, this research emphasizes the importance of exploring viral diversity in uncultivated fungi, bridging knowledge gaps in mycovirus ecology. The discoveries of a novel flexivirus with unique genome organization and endornaviruses in T. claveryi broaden our comprehension of mycovirus diversity and evolution, highlighting the need for continued investigations into viral populations in wild fungi.
Asunto(s)
Virus Fúngicos , Genoma Viral , Micorrizas , Filogenia , Virus Fúngicos/genética , Virus Fúngicos/clasificación , Virus Fúngicos/aislamiento & purificación , Micorrizas/genética , Micorrizas/virología , Micorrizas/clasificación , Virus ARN/genética , Virus ARN/clasificación , Virus ARN/aislamiento & purificación , ARN Viral/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Basidiomycota/virología , Basidiomycota/genéticaRESUMEN
Fungi harbor a vast diversity of mobile genetic elements (MGEs). Recently, novel fungal MGEs, tentatively referred to as 'ambiviruses,' were described. 'Ambiviruses' have single-stranded RNA genomes of about 4-5 kb in length that contain at least two open reading frames (ORFs) in non-overlapping ambisense orientation. Both ORFs are conserved among all currently known 'ambiviruses,' and one of them encodes a distinct viral RNA-directed RNA polymerase (RdRP), the hallmark gene of ribovirian kingdom Orthornavirae. However, 'ambivirus' genomes are circular and predicted to replicate via a rolling-circle mechanism. Their genomes are also predicted to form rod-like structures and contain ribozymes in various combinations in both sense and antisense orientations-features reminiscent of viroids, virusoids, ribozyvirian kolmiovirids, and yet-unclassified MGEs (such as 'epsilonviruses,' 'zetaviruses,' and some 'obelisks'). As a first step toward the formal classification of 'ambiviruses,' the International Committee on Taxonomy of Viruses (ICTV) recently approved the establishment of a novel ribovirian phylum, Ambiviricota, to accommodate an initial set of 20 members with well-annotated genome sequences.
Asunto(s)
Genoma Viral , Sistemas de Lectura Abierta , Viroides , Viroides/genética , Viroides/clasificación , Filogenia , ARN Viral/genética , Virus ARN/genética , Virus ARN/clasificación , Hongos/genética , Hongos/virología , ARN Polimerasa Dependiente del ARN/genética , Virus Fúngicos/genética , Virus Fúngicos/clasificación , Virus Fúngicos/aislamiento & purificaciónRESUMEN
In the current study, a novel strain of Fusarium oxysporum alternavirus 1 (FoAV1) was identified from the Fusarium oxysporum f. sp. melonis (FOM) strain T-BJ17 and was designated as Fusarium oxysporum alternavirus 1-FOM (FoAV1-FOM). Its genome consists of four dsRNA segments of 3515 bp (dsRNA1), 2663 bp (dsRNA2), 2368 bp (dsRNA3), and 1776 bp (dsRNA4) in length. Open reading frame 1 (ORF1) in dsRNA1 was found to encode a putative RNA-dependent RNA polymerase (RdRp), whose amino acid sequence was 99.02% identical to that of its counterpart in FoAV1; while ORF2 in dsRNA2, ORF3 in dsRNA3, and ORF4 in dsRNA4 were all found to encode hypothetical proteins. Strain T-BJ17-VF, which was verified to FoAV1-FOM-free, was obtained using single-hyphal-tip culture combined with high-temperature treatment to eliminate FoAV1-FOM from strain T-BJ17. The colony growth rate, ability to produce spores, and virulence of strain T-BJ17 were significantly lower than those of T-BJ17-VF, while the dry weight of the mycelial biomass and the sensitivity to difenoconazole and pydiflumetofen of strain T-BJ17 were greater than those of T-BJ17-VF. FoAV1-FOM was capable of 100% vertical transmission via spores. To our knowledge, this is the first time that an alternavirus has infected FOM, and this is the first report of hypovirulence and increased sensitivity to difenoconazole and pydiflumetofen induced by FoAV1-FOM infection in FOM.
Asunto(s)
Virus Fúngicos , Fusarium , Genoma Viral , Enfermedades de las Plantas , Triazoles , Fusarium/efectos de los fármacos , Fusarium/genética , Fusarium/virología , Fusarium/patogenicidad , Virus Fúngicos/genética , Virus Fúngicos/aislamiento & purificación , Virus Fúngicos/clasificación , Virus Fúngicos/fisiología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/virología , Triazoles/farmacología , Dioxolanos/farmacología , Virulencia , Virus ARN/genética , Virus ARN/aislamiento & purificación , Virus ARN/efectos de los fármacos , Virus ARN/clasificación , Filogenia , Sistemas de Lectura Abierta , Triticum/microbiología , Triticum/virologíaRESUMEN
It is extremely rare that a single virus crosses host barriers across multiple kingdoms. Based on phylogenetic and paleovirological analyses, it has previously been hypothesized that single members of the family Partitiviridae could cross multiple kingdoms. Partitiviridae accommodates members characterized by their simple bisegmented double-stranded RNA genome; asymptomatic infections of host organisms; the absence of an extracellular route for entry in nature; and collectively broad host range. Herein, we show the replicability of single fungal partitiviruses in three kingdoms of host organisms: Fungi, Plantae, and Animalia. Betapartitiviruses of the phytopathogenic fungusRosellinia necatrix could replicate in protoplasts of the carrot (Daucus carota), Nicotiana benthamiana and Nicotiana tabacum, in some cases reaching a level detectable by agarose gel electrophoresis. Moreover, betapartitiviruses showed more robust replication than the tested alphapartitiviruses. One of the fungal betapartitiviruses, RnPV18, could persistently and stably infect carrot plants regenerated from virion-transfected protoplasts. Both alpha- and betapartitiviruses, although with different host preference, could replicate in two insect cell lines derived from the fall armyworm Spodoptera frugiperda and the fruit fly Drosophila melanogaster. Our results indicate the replicability of single partitiviruses in members of three kingdoms and provide insights into virus adaptation, host jumping, and evolution.
Asunto(s)
Daucus carota , Nicotiana , Replicación Viral , Animales , Nicotiana/virología , Nicotiana/microbiología , Daucus carota/virología , Daucus carota/microbiología , Virus ARN/genética , Virus ARN/fisiología , Virus Fúngicos/genética , Virus Fúngicos/clasificación , Virus Fúngicos/fisiología , Filogenia , Protoplastos/virología , Enfermedades de las Plantas/virología , Enfermedades de las Plantas/microbiología , Spodoptera/virología , Spodoptera/microbiologíaRESUMEN
In recent years, numerous oomycete mycoviruses have been discovered; however, very few studies have focused on their effects on the host oomycete phenotype. In this study, we investigated the impact of toti-like Pythium ultimum RNA virus 2 (PuRV2) infection on the phytopathogenic soil-borne oomycete Globisporangium ultimum, which serves as a model species for Globisporangium and Pythium, specifically the UOP226 isolate in Japan. We generated a PuRV2-free isogenic line through hyphal tip isolation using high-temperature culture and subsequently compared the phenotypic characteristics and gene expression profiles of UOP226 and the PuRV2-free isogenic line. Our findings revealed that the metalaxyl sensitivity of UOP226 was greater than that of the PuRV2-free isogenic line, whereas the mycelial growth rate and colony morphology remained unchanged in the absence of the fungicide. Furthermore, transcriptome analyses using RNA-seq revealed significant downregulation of ABC-type transporter genes, which are involved in fungicide sensitivity, in UOP226. Our results suggest that PuRV2 infection influences the ecology of G. ultimum in agricultural ecosystems where metalaxyl is applied.
Asunto(s)
Alanina , Virus Fúngicos , Fungicidas Industriales , Enfermedades de las Plantas , Virus ARN , Fungicidas Industriales/farmacología , Virus Fúngicos/genética , Virus Fúngicos/fisiología , Virus Fúngicos/aislamiento & purificación , Virus Fúngicos/efectos de los fármacos , Alanina/análogos & derivados , Alanina/farmacología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/virología , Virus ARN/efectos de los fármacos , Virus ARN/genética , Pythium/efectos de los fármacos , Pythium/crecimiento & desarrollo , Hifa/crecimiento & desarrollo , Hifa/efectos de los fármacos , Perfilación de la Expresión Génica , Micelio/crecimiento & desarrollo , Micelio/efectos de los fármacos , Micelio/virología , Japón , TranscriptomaRESUMEN
A wide diversity of mycoviruses has been reported from Botrytis species, some with the potential to suppress the pathogenic abilities of this fungus. Considering their importance, this study was devised to find potential hypovirulence-associated mycoviruses found in Botrytis cinerea strains isolated from Pakistani strawberry fields. Here we report the complete genome characterization of two fusariviruses co-infecting a single isolate of phytopathogenic fungus B. cinerea (Kst14a). The viral genomes were sequenced by deep sequencing using total RNA fractions of the Kst14a isolate. The identified viruses were tentatively named Botrytis cinerea fusarivirus 9 (BcFV9) and Botrytis cinerea fusarivirus 3a (BcFV3a). Both viruses had a single-segmented (ssRNA) genome having a size of 6424 and 8370 nucleotides encoding two discontinuous open reading frames (ORFs). ORF-1 of both mycoviruses encodes for a polyprotein having a conserved domain of RNA-dependent RNA polymerase (RdRP) and a helicase domain (Hel) which function in RNA replication, while ORF2 encodes a hypothetical protein with an unknown function, respectively. Phylogenetic analysis indicated that BcFV9 made a clade with the genus Alphafusarivirus and BcFV3a fall in the genus Betafusarivirus in the family Fusariviridae. To our knowledge, this is the first report of two fusariviruses identified in isolates of B. cinerea from Pakistan. Both mycoviruses successfully transfected to a compatible strain of B. cinerea (Mst11). A comparison of virus-free (VF) and virus-infected (VI) isogenic lines showed the presence of these viruses was causing hypovirulence in infected strains. Virus-infected strains also had a small lesion size while testing the pathogenicity via apple assay.
Asunto(s)
Botrytis , Virus Fúngicos , Genoma Viral , Sistemas de Lectura Abierta , Filogenia , Enfermedades de las Plantas , Botrytis/virología , Botrytis/genética , Virus Fúngicos/genética , Virus Fúngicos/aislamiento & purificación , Virus Fúngicos/clasificación , Enfermedades de las Plantas/microbiología , ARN Viral/genética , Fragaria/microbiología , Fragaria/virología , Pakistán , Proteínas Virales/genética , Secuenciación de Nucleótidos de Alto RendimientoRESUMEN
A novel mitovirus was identified in Fusarium oxysporum f. sp. melonis strain T-SD3 and designated as "Fusarium oxysporum mitovirus 3" (FoMV3). The virus was isolated from diseased muskmelon plants with the typical symptom of fusarium wilt. The complete genome of FoMV3 is 2269 nt in length with a predicted AU content of 61.40% and contains a single open reading frame (ORF) using the fungal mitochondrial genetic code. The ORF was predicted to encode a polypeptide of 679 amino acids (aa) containing a conserved RNA-dependent RNA polymerase (RdRp) domain with a molecular mass of 77.39 kDa, which contains six conserved motifs with the highly conserved GDD tripeptide in motif IV. The 5'-untranslated region (UTR) and 3'-UTR of FoMV3 were predicted to fold into stem-loop structures. BLASTp analysis revealed that the RdRp of FoMV3 shared the highest aa sequence identity (83.85%) with that of Fusarium asiaticum mitovirus 5 (FaMV5, a member of the family Mitoviridae) infecting F. asiaticum, the causal agent of wheat fusarium head blight. Phylogenetic analysis further suggested that FoMV3 is a new member of the genus Unuamitovirus within the family Mitoviridae. This is the first report of a new mitovirus associated with F. oxysporum f. sp. melonis.
Asunto(s)
Virus Fúngicos , Fusarium , Genoma Viral , Sistemas de Lectura Abierta , Filogenia , Enfermedades de las Plantas , Fusarium/virología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/virología , Virus Fúngicos/genética , Virus Fúngicos/aislamiento & purificación , Virus Fúngicos/clasificación , Virus ARN/genética , Virus ARN/aislamiento & purificación , Virus ARN/clasificación , Secuenciación Completa del Genoma , ARN Viral/genética , ARN Polimerasa Dependiente del ARN/genética , Proteínas Virales/genética , Cucumis melo/virología , Cucumis melo/microbiología , Secuencia de Aminoácidos , Regiones no Traducidas 5' , Regiones no Traducidas 3' , Secuencia de BasesRESUMEN
A novel negative-sense single-stranded RNA mycovirus, designated as "Magnaporthe oryzae mymonavirus 1" (MoMNV1), was identified in the rice blast fungus Magnaporthe oryzae isolate NJ39. MoMNV1 has a single genomic RNA segment consisting of 10,515 nucleotides, which contains six open reading frames. The largest open reading frame contains 5837 bases and encodes an RNA replicase. The six open reading frames have no overlap and are arranged linearly on the genome, but the spacing of the genes is small, with a maximum of 315 bases and a minimum of 80 bases. Genome comparison and phylogenetic analysis indicated that MoMNV1 is a new member of the genus Penicillimonavirus of the family Mymonaviridae.