• 식물병연구
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• 제12권1호
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• pp.62-64
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• 2006

2005년 7월 전북고창 지역에서 옥수수에 줄무늬 위축증상을 나타내는 이병주를 채집하여 게놈 dsRNA 분리 및 RT-PCR 반응을 실시하였다. 게놈 dsRNA 분리는 이병엽 100mg에서 직접 dsRNA를 추출하여 agarose gel에서 전기영동 하였으며, 10개의 분절을 확인하였다. 추출한 dsRNA를 주형으로 하여 S7, S8, S10 full-length 특이적인 primer를 제작하여 RT-PCR을 실시하였다. 그 결과 각각의 분절에서 예상되는 크기의 밴드를 확인하였다. 고창지역의 사료용 옥수수 재배지에서 RBSDV의 발생면적은 약 22ha에 달하였으며 발생이 심한 곳은 약 80%의 이병율을 나타내기도 하였다.

• The Plant Pathology Journal
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• 제21권2호
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• pp.172-173
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• 2005

Until now, occurrence of Rice black-streaked dwarf virus (RBSDV) is observed in Gyeongsang provinces, southeastern part of Korea. However, recently, the occurrence of RBSDV is increasing and spreading in Jeonra provinces including Gochang-gun, southwestern part of Korea. RBSDV infected plants showed typical symptoms including stunted, deformed leaves with white waxy or black-streaked swelling along the veins. We extracted viral genomic dsRNA from infected leaves and analyzed dsRNA pattern by polyacrylamide gel electrophoresis. Ten genomic segments with similar sized dsRNAs were observed. We also detected RBSDV by reverse transcription (RT)-PCR using specific primers for S10 from genomic dsRNA and observed amplified DNA fragment specific for RBSDV S10.

• 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

• 미생물학회지
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• 제29권4호
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• pp.221-225
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• 1991

The RNA transcripts produced from in vitro transcription reaction of BTV core were analyzed on agarose-urea gel. Fast migrating abortive RNAs, in addition to full length species of RNA, were observed. Fast migrating RNAs extracted from agarose-urea gel were hybridized to all 10 segments of genomic ds RNA, while solw migrating RNAs extracted from agarose-urea gel were hybridized only to the large and medium size genomic ds RNA. These results indicate that fast migrating RNA transcripts are most likely the products of abortive transcription.

• The Plant Pathology Journal
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• 제21권4호
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• pp.343-348
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• 2005

The partial nucleotide sequences of the genomic dsRNA mycoviruses infecting Pleurotus ostreatus (isolates ASI2596, ASI2597, and Bupyungbokhoe) and Agaricus blazei Murrill were determined and compared with those of the other dsRNA mycoviruses. Partial nucleotide sequences of the purified dsRNA from ASI2596 and ASI2597 revealed RNA-dependent RNA polymerase sequences that are closely related to Oyster mushroom isometric virus 2, while nucleotide sequences and the deduced amino acid sequence from dsRNA mycovirus infecting Agaricus blazei did not show any significant homology to the other dsRNA mycoviruses. Specific primers were designed for RT-PCR detection of these dsRNA viruses and were found to specifically detect each dsRNA virus. Northern blot analysis confirmed the homogeneity of RT-PCR products to each purified dsRNA. Altogether, our results suggest that these virus-specific primer sets can be employed for the specific detection of each dsRNA mycovirus in infected mushrooms.

• 미생물학회지
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• 제31권3호
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• pp.184-188
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• 1993

Novel Ustilagomaydis strains, designated as SH1 to 14 containing new types of ds RNA segments, are identified from corn smut in Korea. Among 14 isolates, 7 isolates appear to posses virus particles and the other isolates may contain dsRNA as a plasmid form. The pattern of dsRNA is highly diverse form a typical P-type containing one or more of H, M, and L dsRNAs to the one containing one or move M dsRNAs. It is likely that the strains containing H dsRNA posses virus particles which were confirmed by sucrose density gradient followed with different range of specificity and the activity of the strain (SH14) is stronger than A4 toxin. The sensitivity of 14 isolates is also very diverse and two strains (SH10, SH11) appear tobe universal sensitve strains against 5 tested toxin samples.

• 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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• 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).

• 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.

• 미생물학회지
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• 제33권2호
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• pp.105-110
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• 1997

한국에서 분리한 U. maydis에서 바이러스 혹은 dsRNA를 가지고 있는 A series와 SH series를 조합하여 옥수수에서 인위적으로 mating시켜 A45, A23, A211, A310, SH614, SH24의 6개 교배형 균주를 분리하였다. 새로운 교배형 균주에서 바이러스 dsRNA를 비교 분석하여 본 결과, mating전 양친형 균주에서 보이고 있는 H, M, L strand의 dsRNA 양상을 모두 가지고 있고 균주간 특별한 molecular exclusion 현상은 보이지 않았다. 교배형 6개으 균주에서 바이러스를 분리하였다. 분리된 바이러스로부터 dsRNA를 분석한 결과 균주간 mating시 dsRNA 재조합을 통한 새로운 encapsidation은 발생하지 않는다는 것을 확인하였다. Toxin test 결과, SH614는 SH9, SH10, SH11 균주에 대해서, A310과 SH24의 2종은 SH11 균주에 대해 특이적 성장억제 현상을 보였다. 이는 mating에 따른 dsRNA의 재조합이 toxin gene의 발현에 영향을 미치지 않는다는 것을 시사한다.