• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.23-23
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• 2003

Chestnut blight, caused by Cryphonectria parasitica, is the most destructive disease of American and European chestnut trees. A total of 672 C prasitica was isolated from blight lesion on chestnut twigs, which were collected from major chestnut plantations all over Korea in 1999. Isolation rates were over 30% in Kyunggj-, Kyongnam-, and Chonnam-do. The highest isolation rate was 37.4% and recorded in Kyongnam-do. On the other hand, Chonbuk-do had the lowest isolation rate as 13.5%. In grouping of C parasitica by colony shape and color, yellow colony with irregular margin were the most dominant colony type with a frequency of 65.2%. When the 672 isolates were inoculated on the chestnut twigs, 380 isolates (56.5%) caused lesions larger than the standard virulent isolate EP155-2, while 158 isolates (23.4%) caused smaller lesions than the standard hypovirulent isolate UEP-1. In Bavendamm test that determines phenol oxidase activity, 97.1% of all the isolates resulted the same or darker discoloration than EP155-2, and only 12.2% resulted the same or lighter discoloration than UEP-1. In the vegetative compatibility (VC) tests, total 670 isolates were divided into 121 VC groups (VCGs). Kyongnam-, Chonnam-, and Chungnam-do, the three principal chestnut plantation area, had 49, 33, and 27 VCGs, respectively. Among the VCGs, the biggest VCG, KR-VC104, was composed of 164 isolates and the second biggest VCG had 62 isolates. But, 64 of 121 VCGs consisted of sole member. More than 65.8% of KR-VC104, was isolated from the three provinces, Kyongnam-, Kangwon-, and Chungbuk-do. In KR-VC104, 62.8%, 59.1%, and 85.9% of the isolates looked like virulent in colony type, pathogenicity test, and Bavendamm test. In ds-RNA detection tests using cellulose chromatography, 77 of total 650 isolates were ds-RNA positive and detected ds-RNA segments were approximately 12kb, 3kb, 2.7kb, 2kb, and 1.8kb in size. Among the 77 isolates, 46 isolates had 12kb and 25 isolates had 12kb and 2.7kb. Other 6 Isolates had small ds-RNA segments. Kyongnam-, Chonnam-, and Chungnam-do had 43, 16, and 5 ds-RNA positive isolates, respectively. Among the 121 VCGs, only 29 VCGs had ds-RNA positive isolates.(중략)

• 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.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.351-355
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• 2005

In the storage roots of sweetpotato (Ipomoea batatas (L.) Lam. cv. Kokei 14), 10 to 20% of starch is essentially unbranched linear amylose and the other major component is branched amylopectin. Amylose is produced by the enzyme GBSSI (granule bound starch synthase I), whereas amylopectin is produced by a concerted action of soluble starch synthase and starch branching enzymes (SBEI and SBEII). We constructed double-stranded RNA (dsRNA) interference vectors of GBSSI and IbSBEII and introduced them into sweetpotato genome via Agrobacterium-mediated gene transformation. The endogenous GBSSI expression was inhibited by dsRNA of GBSSI in 73 % of transgenic plants giving rise to the storage tubers containing amylopectin but not amylose. On the other hand, all sweetpotato plants transformed with dsRNA of IbSBEII contained a larger amount of amylose than the non-transgenic control (up to 25% compared to 10% in the controls). The RNA interference (RNAi) is effectively inhibited the gene expression in thestarch metabolic pathway and modified the characteristics of starch in sweetpotato.

• 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).

• Journal of Microbiology and Biotechnology
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• 제31권4호
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• pp.529-539
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• 2021

The beet armyworm, Spodoptera exigua, is a serious insect pest infesting various vegetable crops. Two infectious insect viruses, baculovirus and iflavirus, are known to induce epizootics in S. exigua populations. Indeed, some laboratory colonies have appeared to be covertly infected by these viruses. Diagnostic PCR tests detected two different viruses: Spodoptera exigua multiple nucleopolyhedrosis virus (SeMNPV) and iflaviruses (SeIfV1 and SeIfV2). Viral extract from dead larvae of S. exigua could infect Sf9 cells and produce occlusion bodies (OBs). Feeding OBs to asymptomatic larvae of S. exigua caused significant viral disease. Interestingly, both SeIfV1 and SeIfV2 increased their titers at late larval stages. Sterilization of laid eggs with 1% sodium hypochloride significantly reduced SeMNPV titers and increased larval survival rate. Double-stranded RNA (dsRNA) specific to SeIfV1 or SeIfV2 significantly reduced viral titers and increased larval survival rate. To continuously feed dsRNA, a recombinant Escherichia coli HT115 expressing SeIfV1-dsRNA was constructed with an L4440 expression vector. Adding this recombinant E. coli to the artificial diet significantly reduced the SeIfV1 titer and increased larval survival. These results indicate that laboratory colony collapse of S. exigua is induced by multiple viral infections. In addition, either suppression of SeMNPV or SeIfV infection significantly increased larval survival, suggesting a cooperative pathogenicity between baculovirus and iflavirus against S. exigua.

• 한국버섯학회지
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• 제8권2호
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• pp.41-50
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• 2010

Mycoviruses have been found in many fungal species including mushrooms. Double-stranded (ds) RNA genomes were common type in mycoviruses, but single-stranded (ss) RNA mycoviruses were also reported in some fungal species. Sequencing analysis using cDNA cloning experiments revealed that mycoviruses can be classified into several different virus families such as Totiviridae, Hypoviridae, Partitiviridae and Barnaviridae etc. Because the nucleotide sequence data that are available in these days are very limited in a number of mycoviruses, the existence of more diverse viral groups in fungi are currently expected. In this review, we selected four different fungal groups, which were considered as the model systems for mycovirus related studies in both plant pathogenic fungi and edible mushroom species, and discussed about their molecular characteristics of diverse mycoviruses. The plant pathogenic fungi introduced here were Cryphonectria parasitica and Helminthosporium victoriae and the edible mushroom species were Agaricus bisporus and Pleurotus ostreatus.

• KSBB Journal
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• 제26권2호
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• pp.131-138
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• 2011

Some computational approaches are needed for clarifying RNAi sequences, because it takes much time and endeavor that almost of RNAi sequences are verified by experimental data. Incorrectness of RNAi mechanism and other unaware factors in organism system are frequently faced with questions regarding potential use of RNAi as therapeutic applications. Our massive parallelized pair alignment scoring between dsRNA in Genebank and expressed sequence tags (ESTs) in Caenorhabditis elegans Genome Sequencing Projects revealed that this provides a useful tool for the prediction of RNAi induced cosuppression details for practical use. This pair alignment scoring method using high performance computing exhibited some possibility that numerous unwanted gene silencing and cosuppression exist even at high matching scores each other. The classifying the relative higher matching score of them based on GO (Gene Ontology) system could present mapping dsRNA of C. elegans and functional roles in an applied system. Our prediction also exhibited that more than 78% of the predicted co-suppressible genes are located in the ribosomal spot of C. elegans.

• 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.

• 생명과학회지
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• 제19권3호
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• pp.317-321
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• 2009

다운증후군은 추가적으로 존재하는 인간염색체 21번에 위치한 유전자의 과발현으로 발병한다. 다운증후군 환자에서 보이는 여러 증상들 중 학습과 기억능력 저하와 같은 뇌기능 저하는 다운증후군 환자가 독립적인 생활을 영위하는데 가장 큰 걸림돌이 된다. 인간염색체 21번에 위치하는 Dyrk1A는 신경발달에 중요한 역할을 하는 단백질로 Dyrk1A를 과발현 하는 형질전환 생쥐에서 심각한 해마 의존적 학습과 기억 장애가 보고되었다. 본 연구에서는 인간 Dyrk1A를 과발현 하는 형질전환 생쥐와 RNA interference (RNAi) 방법을 이용하여 endogenous mouse Dyrk1A의 발현은 정상적으로 유지하면서 exogenous human Dyrk1A 발현은 특이적으로 저해함으로써 인간 Dyrk1A 과발현에 의한 학습과 기억 능력저하를 회복시킬 수 있는지 동물모델에서 검증하기 위한 첫 단계로 인간 Dyrk1A 특이적 lentiviral short hairpin RNA (shRNA)를 발굴하였다. 발굴된 shRNA를 이용한 형질전환 모델생쥐에서의 증상의 회복 가능성 검증은 다운증후군의 뇌기능저하 치료제 개발에 중요한 정보를 제공할 것이다.

• Bulletin of the Korean Chemical Society
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• 제32권11호
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• pp.3893-3898
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• 2011

Cellular uptake of double-stranded DNA (dsDNA) triggers strong innate immune responses via activation of NF-${\kappa}B$ transcription factor. However, the detailed mechanism of dsDNA-mediated innate immune response remains yet to be elucidated. Here, we show that the expression of tazarotene-induced gene 3 (TIG3) is dramatically induced by dsDNA stimulation, and the siRNA-mediated down-regulation of TIG3 mRNA results in significant suppression of dsDNA-triggered cytokine expression. Because TIG3 has been previously shown to physically interact with transglutaminase (TG) 1 to activate TG activity, and TG2 has been shown to induce NF-${\kappa}B$ activity by inducing $I{\kappa}B{\alpha}$ polymerization, we tested whether TG also plays a role in dsDNA-mediated innate immune response. Pre-treatment of TG inhibitors dramatically reduces dsDNA-triggered cytokine induction. We also show that, in HeLa cells, TG2 is the major TG, and TIG3 physically interacts with TG2. Combined together, our results suggest a novel mechanism of dsDNA-triggered innate immune response which is critically dependent on TIG3 and TG2.