• The Plant Pathology Journal
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• 제27권3호
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• pp.285-290
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• 2011

Fusarium graminearum virus 2 (FgV2) infects Fusarium graminearum strain 98-8-60 and has at least five segments of double-stranded RNAs (dsRNAs), denoted as dsRNA-1 to dsRNA-5. In this study, the genome of FgV2 was sequenced and its phylogenetic relationship with other mycoviruses was analyzed. The lengths of FgV2 dsRNAs 1-5 ranged from 2414 to 3580 base pairs (bp). The 5' and 3' untranslated regions (UTRs) are highly conserved, and each dsRNA segment had 78-105 and 84-306 bp of 5' and 3' UTRs, respectively. Each dsRNA segment contained a single open reading frame (ORF). Computer analysis of dsRNA-1 revealed a putative open reading frame (ORF) that shows high sequence identity with an RNA-dependent RNA polymerase (RdRp) containing eight conserved motifs. dsRNAs 2-5 also each contain one putative ORF coding for products of unknown function. The sequences of FgV2 dsRNA-2 and dsRNA-3 have significant sequence identity with Magnaporthe oryzae chrysovirus 1 (MoCV1) dsRNA-3 and -4, respectively. When compared to other dsRNA mycoviruses in a phylogenetic analysis of the putative RdRp protein, FgV2 was found to form a distinct virus clade with Aspergillus mycovirus 1816 and MoCV1 in the family Chrysoviridae.

• 한국균학회지
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• 제25권3호통권82호
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• pp.191-201
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• 1997

유전자의 삽입에 의해 발생하는 C. parasitica의 돌연변이체중 mycovirus에 감염된 것과 같이 색소와 포자를 적게 형성하는 균주(HSM1)를 선발하였다. 선발된 균주는 형태학적 병징외에도 laccase효소의 역가와 같은 생화학적 그리고 표지 유전자들을 통해 분자 생물학적인 특징도 virus에 감염된 균주와 동일한 특징을 나타냈다. HSM1에서 돌연변이가 일어날 부위를 cloning하여 조사한 결과, 유전자 삽입 부위는 C. parasitica의 두 유전자(Cpg2와Cpg3)의 사이(intergenic space)이며 유전자의 삽입 결과, HSM1에서 Cpg2의 발현이 오히려 증가됨이 관찰되었고, 나아가 이와 같은 현상은 mycovirus 감염 균주(UEP1)에서도 일어나고 있음을 확인하였다.

• Molecules and Cells
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• 제23권3호
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• pp.304-315
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• 2007

Fusarium graminearum causes a serious scab disease of small grains in Korea. The nucleotide sequence of the genomic RNA of a double-stranded RNA (dsRNA) virus, Fusarium graminearum virus-DK21 (FgV-DK21), from F. graminearum strain DK21, which is associated with hypovirulence in F. graminearum, was determined and compared to the genome sequences of other mycoviruses, including Cryponectria hypoviruses. The FgV-DK21 dsRNA consists of 6,624 nucleotides, excluding the 3'-terminal poly(A) tail. The viral genome has 53- and 46-nucleotide 5' and 3' untranslated regions (UTRs), respectively, and five putative open reading frames. A phylogenetic analysis of the deduced amino acid sequence of ORF1, which encodes a putative RNA-dependent RNA polymerase, and those of other mycoviruses revealed that this organism forms a distinct virus clade with other hypoviruses, and is more distantly related to other mycoviruses (3.8 to 24.0% identity). However, pairwise sequence comparisons of the nucleotide and deduced amino acid sequences of ORFs 2 through 5 revealed no close relationships to other protein sequences currently available in GenBank. Analyses of RNA accumulation by Northern blot and primer extension indicated that these putative gene products are expressed from at least two different subgenomic RNAs (sgRNAs), in contrast to the cases in other hypoviruses. This study suggests the existence of a new, as yet unassigned, genus of mycoviruses that exhibits a potex-like genome organization and sgRNA accumulation.

• The Plant Pathology Journal
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• 제30권2호
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• pp.151-158
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• 2014

Three double-stranded RNAs (dsRNAs), approximately 1.85, 1.65 and 1.27 kb in size, were detected in an isolate of Cytospora sacchari from Iran. Partial nucleotide sequence revealed a 1,284 bp segment containing one ORF that potentially encodes a 405 aa protein. This protein contains conserved motifs related to RNA dependent RNA polymerases (RdRp) that showed similarity to RdRps of partitiviruses. The results indicate that these dsRNAs represent a novel Partitivirus that we tentatively designate Cytospora sacchari partitivirus (CsPV). Treatment of the fungal strain by cyclohexamide and also hyphal tip culture had no effect on removing the putative virus. Phylogenetic analysis of putative RdRp of CsPV and other partitiviruses places CsPV as a member of the genus Partitivirus in the family Partitiviridae, and clustering with Aspergillus ochraceous virus 1.

• International journal of advanced smart convergence
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• 제8권3호
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• pp.115-122
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• 2019

Recent and ongoing discoveries of mycoviruses with new properties demand the development of an appropriate research infrastructure to analyze their evolution and classification. In particular, the discovery of negative-sense single-stranded mycoviruses is worth noting in genome types in which double-stranded RNA virus and positive-sense single-stranded RNA virus were predominant. In addition, some genomic properties of mycoviruses are more interesting because they have been reported to have similarities with the pathogenic virus family that infects humans and animals. Genetic information on mycoviruses continues to accumulate in public repositories; however, these databases have some difficulty reflecting the latest taxonomic information and obtaining specialized data for mycoviruses. Therefore, in this study, we developed a bioinformatics-based pipeline to efficiently utilize this genetic information. We also designed a schema for data processing and database construction and an algorithm to keep taxonomic information of mycoviruses up to date. The pipeline and database (termed 'mycoVDB') presented in this study are expected to serve as useful foundations for improving the accuracy and efficiency of future research on mycoviruses.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.117.3-118
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• 2003

We tested for the presence of double-stranded RNA (dsRNA) mycovirus in 827 Fusarium graminearum isolated from diseased barley and maize. dsRNA mycoviruses with various sizes were isolated. Of them, it was previously reported that dsRNA from DK2l isolate had pronounced morphological changes, including reduction in mycelial growth, increased to red pigmentation, reduced virulence and sporulation. (Chu et al., Appl. Environ. Microbiol. 2002). For better understanding of this hypovirulence associated with DK2l dsRNA virus, we determined the complete nucleotide sequence of dsRNA genome and named Fusarium hypovirus DK2l strain (Fhv-DK2l ). Genomic RNA of Fhv-DK2l was determined to be 6625 nucleotides in length excluding the poly (A) tail and contained three putative open reading frame. RNA-dependent RNA polymerase (RdRp) and helicase domain were expected in ORF A, 54 to 4709 nucleotide position. ORE B, 4752 to 5216 nucleotide position, and ORF C, 5475 to 6578 nucleotide position, were predicted to encode 16.7kDa and 41.3kDa protein respectively each. We could not detect any conserved domains from these two proteins. Phylogenetic analysis showed Fhv-DK2l was related to Cryphonectria hypovirus 3. Ten additional isolates were found that were infected with dsRNA mycoviruses. These mycoviruses contain 2 to 4 different segments of dsRNAs with the size range of approximately 1.7 to 10-kbp in length. The presence of dsRNAs isolates did not affect colony morphology and were transmissible through conidia and ascospore with incidence of 30-100％. These results indicate that there is genomic diversity of dsRNA mycoviruses that infect F. graminearum isolates and that impact of virus infection on host's morphology and virulence is determined by the interaction between dsRNAs and the fungal host, not by the mere presence of the dsRNAs

• The Plant Pathology Journal
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• 제22권1호
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• pp.68-74
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• 2006

A new virus with a dsRNA genome was isolated and characterized from the Suhan-:neutari strain (ASI 2504) of Pleurotus ostreatus, which was characterized as long and slightly bent with small caps on the stipe of fruit body. Thirty nm isometric viruses with three dsRNA segments (approximately 2.0, 1.84 and 1.82 kb in sizes) were isolated by ultracentrifugation in sucrose gradients. Western analysis of protein extracted purified viruses with anti-virus polyclonal antibody confirmed that viruses have two specific proteins (36 and 68 kDa). Computer analysis of 2.0 kb segment shows that high. sequence identity with RNA-dependent RNA polymerase (RdRp) of partitiviruses, respectively. When compared to other dsRNA mycoviruses in a phylogenetic analysis, OMDV was most related to Pleurotus ostreatus virus 1.

• The Plant Pathology Journal
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• 제16권3호
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• pp.142-146
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• 2000

Ordered differential display using RT-PCR (ODD-PCR) was conducted to have a profile of the differently expressed genes between a hypovirulent strain of Cryphonectria parasitica (UEP1) and its isogenic wild type strain (EP155/2). ODD-PCR has advantages of high sensitivity, reproducibility, proportional representation, and limited number of primer combinations comparing with other differential display methods. RNAs were prepared from 1 and 5 day liquid culture of both hypovirulent and wild type strains, and were further evaluated with the marker genes of C. parasitica such as cryparin and mating factor MF2-1, which were already proven to be specifically down-regulated by the presence of mycovirus CHV1-713. ODD-PCR was conducted using those RNAs and expressed genes were categorized to five groups according to their temporal and quantitative expression patterns. Those fives groups are CPC, CPE, CPL, CPD, and CPU which represent constitutively-expressed, early-expressed, late-expressed, down-regulated, and up-regulated, respectively. Ninety two primer combinations out of a total of 192 have been tested so far. Among the twenty to fifty distinct bands per each reaction, an average of four to ten genes was identified as viral-regulated fungal genes. Those viral-specifc genes were further analyzed by DNA sequencing followed by homology search. Characterization of 30 clones including all five groups were conducted as a preliminary data and more are under investigation.

• 한국균학회지
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• 제50권4호
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• pp.263-274
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• 2022

표고는 중요한 상업용 버섯이며, 표고에서의 바이러스 감염에 대한 여러 보고가 있었다. 톱밥 재배 품종인 산백향(NIFoS 2778)과 태향고(NIFoS 4317)에 대한 바이러스 검출 결과, 이들이 2개의 mycoviruses (LeV-HKB 및 LeNSRV1)에 감염되어 있음을 확인하였다. 각 품종의 자실체에서 분리된 담자포자에서 유래한 80개의 단핵균사에서 바이러스 감염을 조사한 결과, 대부분의 단 핵균사들이 바이러스에 감염되어 있었으며, LeV-HKB와 LeNSRV1의 수직감염률에 차이가 있었다. LeV-HKB가 LeNSRV1 보다 높은 수직감염률을 나타낸 것이다. 따라서 mycovirus의 수직 감염 기작이 바이러스 종에 따라 다른 것으로 보인다. 다음으로, 담자포자 유래 단핵균사들의 생장속도를 조사하여 바이러스 감염과 생장속도의 상관관계를 조사하였다. 바이러스 감염과 균사 생장속도 사이에 통계적으로 유의한 상관관계가 없었지만, 감염된 바이러스의 종류가 늘어날수록 생장속도가 감소하는 경향을 확인하였다. 본 연구는 mycovirus의 수직 감염 기작을 이해하는 데 기여하고, 바이러스에 감염되지 않은 단핵균사를 이용한 무바이러스 품종 개발을 촉진하는 데 기여할 수 있을 있을 것 기대된다.