• 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.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.150.3-151
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• 2003

To analyze the genome of Cucumber mosaic virus(CMV) in pepper, we developed a new extraction method for double-stranded RNA(dsRNA). To isolate the dsRNA, 0.1g of pepper leaves homogenized with 1ml of 5${\times}$EXB extraction buffer[0.5M glycin, 0.5M NaCl, 5mM EDTA(pH9.0/NaOH), 10％ Sodium N-lauryl salcosinate(NLS), 10％ Sodium dodecylsulfate(SDS)] and purified with the 1/4 volume of phenol： chloroform： isoamylalcohol(25：24：1). dsRNAs from the aqueous phase was precipitated with isopropanol. This procedure was able to detect a minimal amount of dsRNA from CMV infected plant tissue and to distinguish different CMV satellite RNAs by polyacrylamide gel electrophoresis(PAGE). Moreover, this method can be applied CMV infected in pepper or Rice dwarf virus (RDV) infected rice.

• Journal of Microbiology and Biotechnology
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• 제6권5호
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• pp.361-363
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• 1996

The terminal regions of the double-stranded RNA (dsRNA) genome segments of infectious pancreatic necrosis virus (IPNV) DRT strain were sequenced. The dsRNAs, which were $^{32}P$-labelled at their 3'-termini by incubation with [$^{32}P$]pCp and T4 RNA ligase, were separated by 5$%$ polyacrylamide gel electrophoresis, and the segments A and B of IPNV-DRT were sequenced by two-dimensional gel electrophoresis. The 5'-terminal sequences of the IPNV-DRT plus strand from two genome segments were found to have the same conserved nucleotide (5'-CGG(C/A)A-), but the 3'-terminal sequences -CCCCAGGCG-3' and -CGGACCCCG-3' were found in the plus strand from segments A and B, respectively. The inverted oligonucleotide sequences of 3'-terminal of between segments A and B were found and they differ from those of other IPNVs.

• 식물병연구
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• 제14권3호
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• pp.223-225
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• 2008

2005년 전북 고참에서 채집한 옥수수 이병주로부터 벼검은줄오갈병 바이러스를 동정하였다. 이들 이병주로부터 게놈 dsRNA를 추출하여 polyacrylamide gel 전기영동으로 게놈 패턴을 분석 하였다. 전기영동 결과 이미 알려진 10개의 분절게놈을 확인하였으며 채집지역별 isolate에서 게놈 dsRNA이동도의 차이를 확인하였다. 추출된 dsRNA를 주형으로 하여 S10의 full-length 특이 primer를 사용하여 RT-PCR한 결과 1,801의 예상되는 band를 확인하여 RBSDV로 동정하였다. S10을 pGEM-T vector에 크로닝하여 염기서열 분석 결과 1,801nt, 559aa로 구성되어 있었다. 이는 벼에 발생하는 RBSDV S10의 크기와 동일하였으며 상동성 분석결과 18개 염기에서 변이가 확인되어 99%의 상동성을 나타내었다.

• 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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• 제20권2호
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• pp.155-157
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• 2004

We isolated Rice dwarf virus (RDV) from infected plants in rice fields (Korea, Japan, China, the Philippines and Nepal) and analyzed their genomic dsRNAs by polyacrylamide gel eletrophoresis. The genomic dsRNAs of the isolates showed distinct electrophoretic mobility profiles. The S12 coding to nonstructural protein of Korean isolate (RDV-Kr) was further analyzed by sequencing. The S12 of RDV-Kr was 1,066bp long and coded for a protein composed of 312 amino acids including three open reading frames of P12, P120Pa and P120Pb. The sequence identities were 96% and 98.6% with Japanese isolates (H, AN), 94.7% with Nepalese isolate (NEL), 94% with Chinese isolate (CK) and the Philippines isolate (P).

• Applied Biological Chemistry
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• 제34권2호
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• pp.86-93
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• 1991

CDDP는 $100\;{\mu}M$ 이하의 농도에서 BTV 캡시드 단백질에 crosslink를 형성하지 않는다. 밀도구배초원심분리 결과에 의하면 캡시드 단백질과 RNA사이의 crosslink도 일어나지 않는다. CDDP를 처리한 BTV RNA를 폴리아크릴아마이드 겔에 전기연동하면 RNA이동 패턴이 변한다. 이 결과는 BTV core에 없는 RNA중합효소의 불활성화가 CDDP에 의한 BNA RNA의 구조변화에 의함을 가리킨다.

• The Plant Pathology Journal
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• 제29권4호
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• pp.460-464
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• 2013

dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed identical sequences sequence to known RNA-dependent RNA polymerase genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny.

• Journal of Microbiology and Biotechnology
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• 제4권4호
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• pp.264-269
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• 1994

To determine the nucleotide sequence of the ds RNA segment B containing the RNA dependent RNA polymerase (RdRp) gene of the DRT strain of infectious pancreatic necrosis virus (lPNV), the cDNA of the ds RNA segment B of the DRT strain of IPNV was synthesized using the reverse transcriptase (RT)-polymerase chain reaction (PCR) and its cDNA nucleotide sequence was determined. The DRT segment B was 2, 783 bp long and contained only a single long open reading frame (ORF) of 2, 535 bp in length. This ORF nucleotides encoded the VPl protein, the putative RdRp of IPNV. The VPl protein comsisted of 845 amino acids. The molecular weight of the RdRp, as deduced from the nucleotide sequence, is 94, 426. The nucleotide sequence of the ORF of the DRT showed 89.7% homology to the Jasper strain, but 80.8% to the Sp strain. The amino acid sequence of the ORF of the DRT sho.wed 97.6% homology to the Jasper strain, but 88.7% to the Sp strain. The conserved GTP-binding motif was detected in VPl protein.

• 식물병연구
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• 제14권3호
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• pp.229-232
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• 2008

모자이크 증상의 과꽃(Callistephus chinensis L.)으로부터 Cucumber mosaic virus (CMV)의 한 계통(Cas-CMV)를 분리하고, Fny-CMV와 As-CMV를 대조로 기주반응, dsRNA, RT-PCR 및 RFLP분석을 통하여 바이러스를 동정하였다. Cas-CMV의 특징적인 기주반응의 차이는 박과 식물에서 발현되는 강한 병정이었으며, 특히 쥬키니호박에 접종하였을 때에는 접종 15-20일 후에 심한 모자이크 증상과 함께 어린 식물이 고사되는 괴저현상을 나타냈다. DsRNA분석과 RT-PCR실험의 결과는 Cas-CMV가 서브그룹 I의 CMV에 속하는 것으로 나타났으며, 더욱이 HindIII를 이용한 RFLP 분석은 Cas-CMV가 서브그룹 IA 구분되었다.