• BMB Reports
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• 제41권10호
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• pp.739-744
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• 2008

Turnip yellow mosaic virus (TYMV) is a positive strand RNA virus that infects mainly Cruciferae plants. In this study, the TYMV genome was modified by inserting an extra subgenomic RNA promoter and a multiple cloning site. This modified TYMV was introduced into Nicotiana benthamiana using a Agrobacterium-mediated T-DNA transfer system (agroinfiltration). When a gene encoding $\beta$-glucuronidase or green fluorescent protein was expressed using this modified TYMV as a vector, replication of the recombinant viruses, especially the virus containing $\beta$-glucuronidase gene, was severely inhibited. The suppression of replication was reduced by co-expression of viral silencing suppressor genes, such as tombusviral p19, closteroviral p21 or potyviral HC-Pro. As expected, two subgenomic RNAs were produced from the recombinant TYMV, where the larger one contained the foreign gene. An RNase protection assay revealed that the recombinant subgenomic RNA was encapsidated as efficiently as the genuine subgenomic RNA.

• 식물병연구
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• 제23권1호
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• pp.60-64
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• 2017

무(Raphanus sativus L.) 육종 계통들에 대한 순무모자이크 바이러스(Turnip mosaic virus, TuMV) 저항성을 평가하기 위하여, 20개 무 육종계통들의 잎에 순무모자이크바이러스 BR 병원성을 가지는 국내 분리 계통을 즙액 접종하였다. 순무모자이크바이러스를 접종한 무 개체들은 $22^{\circ}C{\pm}3^{\circ}C$에서 재배하였으며 4주 동안 바이러스 병징 발현을 육안으로 조사하여 저항성을 평가하였다. 순무모자이크바이러스 감염에 의해서 무 육종계통들의 다른 병징 발현에 의해 분석한 결과, 16개 무 계통은 약한 모자이크, 모틀링에서 심한 모자이크 전신 병징을 보였으며 감수성으로 판별되었다. 이러한 결과는 순무모자이크바이러스 외피단백질 유전자에 대한 특이적 역전사중합효소연쇄반응에 의하여 확인되어, 순무모자이크바이러스가 병징을 발현시킨 원인이었다. 이와는 다르게 4개 무 육종계통들에서는 모자이크 등 전신 감염 병징이 발현되지 않았으며, 동일한 무 육종 계통들의 개체들에서 8주 동안 병징이 관찰되지 않았다. 역전사중합효소연쇄반응으로 조사한 결과 4개 무 육종 계통들의 상엽들에서 순무모자이크바이러스가 검출되지 않았다. 이런 결과는 4개 무 육종계통들이 순무모자이크바이러스에 대한 강한 저항성을 가지고 있음을 예시해준다.

• BMB Reports
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• 제47권6호
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• pp.330-335
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• 2014

Turnip yellow mosaic virus (TYMV) is a spherical plant virus that has a single 6.3 kb positive strand RNA as a genome. In this study, RNA1 sequence of Flock house virus (FHV) was inserted into the TYMV genome to test whether TYMV can accommodate and express another viral entity. In the resulting construct, designated TY-FHV, the FHV RNA1 sequence was expressed as a TYMV subgenomic RNA. Northern analysis of the Nicotiana benthamiana leaves agroinfiltrated with the TY-FHV showed that both genomic and subgenomic FHV RNAs were abundantly produced. This indicates that the FHV RNA1 sequence was correctly expressed and translated to produce a functional FHV replicase. Although these FHV RNAs were not encapsidated, the FHV RNA having a TYMV CP sequence at the 3'-end was efficiently encapsidated. When an eGFP gene was inserted into the B2 ORF of the FHV sequence, a fusion protein of B2-eGFP was produced as expected.

• Journal of Microbiology and Biotechnology
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• 제29권11호
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• pp.1790-1798
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• 2019

Flock House virus (FHV), an insect RNA virus, has a bipartite genome. FHV RNA1 can be packaged in turnip yellow mosaic virus (TYMV) as long as the FHV RNA has a TYMV sequence at the 3'-end. The encapsidated FHV RNA1 has four additional nucleotides at the 5'-end. We investigated whether the recombinant FHV RNA1 could replicate in mammalian cells. To address this issue, we prepared in vitro transcribed FHV RNAs that mimicked the recombinant FHV RNA1, and introduced them into baby hamster kidney (BHK) cells. The result showed that the recombinant FHV RNA1 was capable of replication. An eGFP gene inserted into the frame with B2 gene of the FHV RNA1 was also successfully expressed. We also observed that eGFP expression at the protein level was strong at 28℃ but weak at 30℃. Sequence analysis showed that the 3'-ends of the RNA1 and RNA3 replication products were identical to those of the authentic FHV RNAs. This indicates that FHV replicase correctly recognized an internally-located replication signal. In contrast, the 5'-ends of recombinant FHV RNA1 frequently had deletions, indicating random initiation of (+)-strand synthesis.

• 식물병과 농업
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• 제1권1호
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• pp.45-46
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• 1995

In recent years, there were considerably severe occurrences of TuMV(turnip mosaic virus) disease on radish and Chinese cabbage cultivated at alpine or sub-alpine regions, especially more severe on young Chinese cabbage sowed after late June. Started from 1991, those were very severe in 1992 and 1994, for the number of migrated aphids was increased enormously according to the weather condition of high temperature and low humidity then. This disease started at late June to early July, and continued to late August. It seemed that TuMV was transmitted easily and completely to the young chinese cabbages, but hardly and rarely the old. The regions over 1,000m of altitude had less possibility of disease-occurring, but there was severe occurrence on the second cropping of Chinese cabbage in a year. It is considered that more researches on control method of TuMV disease will be needed very urgently.

• BMB Reports
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• 제41권11호
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• pp.778-783
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• 2008

Turnip yellow mosaic virus (TYMV) RNA has two hairpins in its 5' untranslated region (5'-UTR). To investigate the role of the hairpins in replication of TYMV, mutants lacking one or both of the two hairpins were constructed. The TYMV constructs were introduced into Chinese cabbage by an Agrobacterium-mediated T-DNA transfer method, called agroinfiltration. Analysis of total RNA from agroinfiltrated leaves showed that replication of the mutant TYMV RNA lacking both hairpins was about 1/100 of wild type. This mutant was also impaired in systemic spread. Deletion analysis of each hairpin revealed that both hairpins were needed for maximal replication. The deletion analysis along with sequence modification of the hairpin structure indicates that the second hairpin plays a role in efficient long-distance systemic movement of TYMV.

• 한국식물병리학회지
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• 제14권6호
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• pp.625-629
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• 1998

Turnip mosaic potyviruses (TuMV) were isolated from Rorippa indica and Armoracia lapathifolia showing mosaic symptoms in field. Identification of the TuMVs were carried out by host reactions of indicator plants, electron micrograph, serological properties and reverse transcription-poly-merase chain reaction (RT-PCR). Both viruses systemically infected Chenopodium quinoa, Nicotiana clevelandii, Brassica rapa, B. campestris subsp. pekinensis, B. juncea and Raphanus sativus, and developed local infection on inoculated leaves of C. quinoa, C. amaranticola, C. album, N. tabacum cv. Xanthi nc and Gomphrena grobosa. However, the viruses did not infect on N. glutinosa, Cucumis sativus and Vigna unguiculata. The filamentous particles, about 720 nm in length, and inclusion bodies were observed from the infected leaf tissues by dipping on electron microscopy. Crude sap of leaf infected with the viruses was reacted positively with an antiserum of TuMV in agar gel double diffusion. For detection of the viruses, RT-PCR was carried out with TuMV--specfic oligonucleotide primer. The RT-PCR products, a 1,092 bp DNA fragment, were obtained from naturally infected leaves of R. indica and A. lapathifolia. In inoculation test to seven cruciferous weeds with TuMV, infection occurred in Arabis glabra, Barbarea orthoceras, Capsella bursa-pastoris, Draba nomorosa var. hebecarpa, Rorippa cantoniensis and Thlaspi arvense.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2002년도 총회 및 추계학술발표회
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• pp.119.1-119
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• 2002

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2002년도 총회 및 추계학술발표회
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• pp.119.2-120
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• 2002

• 한국식물병리학회지
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• 제13권4호
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• pp.248-254
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• 1997

순무우 모자이크 바이러스 Ca 계통(TuMV-Ca)의 외피 단백질을 대장균 NM522 strain에서 발현시켰다. 발현된 바이러스 단백질은 한천젤 이중확산법, ELISA와 Western blotting을 이용하여 확인하였다. 외피단백질 발현 벡터(pGEX-Tu)의 구축은 IPTG induction site를 지니는 pGEX-KG에 TuMV-Ca 외피단백질 유전자를 결합하였다. 최적 단백질 발현 조건은 pGEX-Tu를 지니는 대장균을 액체 배지 1 ml당 $A_{595}$=0.1/ml의 농도로 접종한 후 2시간 뒤에 IPTG를 최종 농도를 1 mM로 조절하여 induction 시키는 경우였다. 합성된 목적 단백질은 발현 벡터의 특성상 GST (Glutathion S-Transferase) 단백질과 결합된 형태로 약 59 kDa의 단백질이었다. (uMV CP 33 kDa + GST 26 kDa.)