• 한국응용곤충학회지
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• 제13권1호
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• pp.41-45
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• 1974

우리나라 씨감자에 발생하고 있는 잠복성바이러스의 종류와 이들의 감염상태를 조사하기 위하여 감자 원원종포, 원종포 그리고 원예시험장 등에서 수집한 외관상 무병 건전하게 보이는 감자와 외관상 바이러스병징이 뚜렷한 감자를 대상으로 혈청학적방법에 의해 바이러스 검정을 실시했다. 검정대상 바이러스는 Potato Virus X(PVX), Potato Virus S(PVS), Potato Virus M(PVM), 그리고 Potato Virus Y (PVY)의 4종이며, 바이러스의 검정은 van Slogteren의 미량침강법에 따라 실시했다. 결과는 다음과 같다. 1. 씨감자 생산포장에서 수집한 감자잎에서 PVX, PVS, PVM, 그리고 PVY가 검출되었는데 우리나라 감자에서 PVM이 검출되었다는 실험적 보고는 이것이 처음이다. 2. 외관상 건전하게 보이는 씨감자의 거의 $100\%$ PVX와 PVS에 약 $10\%$가 PVX+PVS+PVM에, 약 $4.5\%$가 PVX+PVS+PVY에, 그리고 $1\%$ 정도가 PVX+PVS+PVM+PVY에 각각 혼합감염되어 있었다. 3. PVX와 PVS만이 검출된 식물은 대부분 병징이 음폐되어 있었으나 PVX, PVS와 함께 PVY가 검출된 식물은 대부분 뚜렷한 병징을 나타내고 있었다. 4. 혈청학적 미량탐강법은 육안적인 관찰만으로는 검정이 어려운 PVX, PVS, PVM 및 mild strain의 PVY를 씨감자 생산지에서 대규모의 식물은 대상으로 신속정확히 검정하는데 애우 편리한 방법으로 생각된다.

• The Plant Pathology Journal
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• 제22권4호
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• pp.386-390
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• 2006

Extensive research of the Potato virus X(PVX) has been performed in in vitro transcription system using the bacteriophage T7 promoter. We constructed an efficient T-DNA based binary vector, pSNU1, and modified vectors carrying PVX replicons. The suitability of the construct to transiently express PVX RNA using Agrobacterium tumefaciens was tested by analysis of infectivity in plants. The expressed PVX RNA was infectous and systemically spread in three plant species including Nicotiana benthamiana, N. tabacum cv. Xanthi-nc, and Capsicum annuum cv. Chilsungcho. The PVX full length construct, pSPVXp31, was caused severe mosaic symptoms on N. benthamiana, severe necrotic lesions on C. annuum while milder symptoms and delayed mosaic symptoms were appeared on the systemic leaves on N. tabaccum. RT-PCR analysis confirmed the presence of PVX RNAs on both inoculated and systemic leaves in all three plant species tested. Our results indicated that PVX replicons were efficiently expressed PVX RNA in at least three tested species. Further investigation win be needed to elucidate the mechanism of PVX replication, translation, movement and assembly/disassembly processes.

• The Plant Pathology Journal
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• 제28권1호
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• pp.75-80
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• 2012

$Potato$ $virus$ $X$ (PVX) contains $cis$-acting elements including stem-loop 1 (SL1) RNA at the 5' region; SL1 is conserved among all potexviruses. The SL1 at the positive-sense RNA, SL1(+), is required for PVX RNA replication, cell-to-cell movement, and translation. Previous research demonstrated that SL1(+) RNA also serves as the origin of assembly for encapsidation of PVX RNA. To identify the essential sequences and/or regions for capsid protein (CP) subunit recognition within SL1(+) RNA, we used electrophoretic mobility shift assays (EMSA), UV cross-linking, and yeast three-hybrid analyses. The EMSA and UV cross-linking analyses with PVX CP subunits and RNA transcripts corresponding to the SL1(+) RNA showed that the SL1(+) RNA formed complexes with CP subunits. We also conducted EMSA and yeast three-hybrid analyses with RNAs containing various mutations of SL1(+) RNA elements. These analyses indicated that SL1(+) RNA is required for the interaction with PVX CP and that the RNA sequences located at the loop C and tetra loop of the SL1(+) are crucial for CP binding. These results indicate that, in addition to being important for RNA accumulation, the SL1(+) RNA from the 5' region of the PVX genome is also required for specific binding of PVX CP.

• The Plant Pathology Journal
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• 제16권2호
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• pp.110-117
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• 2000

Potato virus X (PVX-KO) showing mild mosaic and stunting symptoms on potato (Solanum tuberosum) in Kangwon area has been isolated and characterized. EM observation of the purified virus particles showed flexuous rod shape of about 520 nm in length. The coat protein (CP) of the virus had a molecular weight of 31 kDa in SDS-PAGE analysis, and the viral RNA was approximately 6.4 kb in size in denatured agarose gel electro-phoresis. In gel-immunodiffusion tests, it reacted strongly with an antiserum to common PVX from BIOREABAAG (USA). A rabbit antiserum was produced using purified virus and used for routine PVX detection by ELISA. Cultivated potatoes in Kangwon and other areas were frequently infected with PVX-KO. Both Datura stramonium and Nicotiana tabaccum cultivars developed necrotic local lesions 5 days after inoculation, and systemic mosaic symptoms with vein clearing 2 weeks after inoculation. All the features agree with the description of other PVX strains. To confirm and determine PVX strains, reverse transcription-polymerase chain reaction experiment was conducted using specific primers for viral CP. Amplified DNA fragments were cloned and sequenced. Results showed nucleotide sequence homologies of about 88 to 99% to other PVX strains. Based on CP amino acid sequence deduced from nucleotide sequences and host range studies PVX-KO is considered a member of the type X subgroup of PVX.

• 한국응용곤충학회지
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• 제16권2호
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• pp.101-104
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• 1977

감자바이러스X(PVX)의 진단과 동정을 위하여 PVX를 순수분리 순화하여 항혈청을 제조하였다. PVX는 지표식물과 전자현미경으로 순수분리하여 도마도애서 증식한 개량된 방법으로 순화하었다. 순화된 PVX의 순화도는 0.59mg/ml이었으며, 이를 1-2ml씩 7일 간격으로 5회 토끼에 주사하였다. 마지막 주사후 10일에 채혈하여 항혈청을 분리하였다. 제조된 항혈청의 력가는 미량침강법에서 1/1024이었고. 한천내확산법에서도 판명한 반응을 관찰할 수 있었다.

• The Plant Pathology Journal
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• 제24권3호
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• pp.361-364
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• 2008

The complete nucleotide sequences of a Korean isolate of Potato virus X(PVX-Kr) has been determined. Full-length cDNA of PVX-Kr has been directly amplified by long template reverse transcription and polymerase chain reaction(RT-PCR) using virus specific 5'-end primer and 3'-end primer, and then constructed in a plasmid vector. Consecutive subclones of a full-length cDNA clone were constructed to identify whole genome sequence of the virus. Total nucleotide sequences of genome of PVX-Kr were 6,435 excluding one adenine at poly A tail, and genome organization was identical with that of typical PVX species. Comparison of whole genome sequence of PVX-Kr with those of European and South American isolates showed 95.4-96.8% and 77.4-77.9%, in nucleotide similarity, respectively. Sequenced PVX-Kr in this study and twelve isolates already reported could be divided into two subgroups in phylogeny based on their complete nucleotide sequences. Phylogenetic tree analysis demonstrated that PVX-Kr was clustered with European and Asian isolates(Taiwan, os, bs, Kr, S, X3, UK3, ROTH1, Tula) in the same subgroup and South American isolates(CP, CP2, CP4, HB) were clustered in the other subgroup.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.14-14
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• 2003

Potato virus X (PVX), the type member of Potexvirus genus, is a flexuous rod-shaped virus containing a single-stranded (＋) RNA. Infection by PVX produces genomic plus- and minus-strand RNAs and two major subgenomic RNAs (sgRNAs). To understand the mechanism for PVX replication, we are studying the cis- and/or trans-acting elements required for RNA replication. Previous studies have shown that the conserved sequences located upstream of two major sgRNAs, as well as elements in the 5' non-translated region (NTR) affect accumulation of genomic and sg RNAs. Complementarity between sequences at the 5' NTR and those located upstream of two major sgRNAs and the binding of host protein(s) to the 5' NTR have shown to be important for PVX RNA replication. The 5 NTR of PVX contains single-stranded AC-rich sequence and stem-loop structure. The potential role(s) of these cis-elements on virus replication, assembly, and their interaction with viral and host protein(s) during virus infection will be discussed based on the data obtained by in vitro binding, in vitro assembly, gel shift mobility assay, host gene expression profiling using various mutants at these regions.

• The Plant Pathology Journal
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• 제24권4호
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• pp.433-438
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• 2008

Potato virus X (PVX) resistance in potato is one of the best-characterized resistance models, however little is known in pepper. To evaluate the resistance to PVX in Capsicum annuum, a total of eleven pepper accessions were used for resistance screening against two PVX strains, USA and UK3. None of them were resistant against strain UK3, whereas four resistant genotypes were found against strain USA, three of which were further characterized. Two unlinked dominant genes were identified for both genotypes Bukang and Perennial; resistance in the genotype CV3 seemed to be conferred by two complementary dominant genes. These results demonstrated that the resistance to PVX in C. annuum is different from that in potato. This is the first report on genetic analysis of PVX resistance in C. annuum.

• The Plant Pathology Journal
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• 제17권3호
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• pp.115-122
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• 2001

Potato virus X (PVX) is a flexuous rod-shaped virus containing a single plus-strand RNA. Viral RNA synthesis is precisely regulated by regulatory viral sequences and by viral and/or host proteins. RNA sequence element as well as stable RNA stem-loop structure in the 5' end of the genome affect accumulation of genomic RNA and subgenomic RNA (sgRNA). The putative sgRNA promoter regions upstream of the PVX triple gene block (TB) and coat protein (CP) gene were critical for both TB and CP sgRNA accumulation. Mutations that disrupted complementarity between a region at the 5' end of the genomic RNA and the sequences located upstream of each sgRNA initiation site is important for PVX RNA accumulation. Compensatory mutations that restore complementarity restored sgRNA accumulation levels. However, the extent of reductions in RNA levels did not directly correlate with the degree of complementarity, suggesting that the sequences of these elements are also important. Gel-retardation assays showed that the 5' end of the positive-strand RNA formed an RNA-protein complex with cellular proteins, suggesting possible involvement of cellular proteins for PVX replication. Future studies on cellular protein binding to the PVX RNA and their role in virus replication will bring a fresh understanding of PVX RNA replication.

• The Plant Pathology Journal
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• 제28권1호
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• pp.40-48
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• 2012

$Potato$ $virus$ $X$ (PVX) replication is precisely regulated by regulatory viral sequences and by viral and/or host proteins. In a previous study, we identified a 54-kDa cellular tobacco protein that bound to a region within the first 46 nucleotides (nt) of the 5' non-translated region (NTR) of the viral genome. Optimal binding was dependent upon the presence of an ACCA sequence at nt 10-13. To identify host factors that bind to 5' NTR elements including AC-rich sequences as well as stemloop 1 (SL1), we used northwestern blotting and matrixassisted laser desorption/ionization time-of-flight mass spectrometry for peptide mass fingerprinting. We screened several host factors that might affect PVX replication and selected a candidate protein, $Nicotiana$ $tabacum$ WRKY transcription factor 1 (NtWRKY1). We used a $Tobacco$ $rattle$ $virus$ (TRV)-based virus-induced gene silencing (VIGS) system to investigate the role of NtWRKY1 in PVX replication. Silencing of $WRKY1$ in $Nicotiana$ $benthamiana$ caused lethal apical necrosis and allowed an increase in PVX RNA accumulation. This result could reflect the balancing of PVX accumulation in a systemic $N.$ $benthamiana$ host to maintain PVX survival and still produce a suitable appearance of mosaic and mottle symptoms. Our results suggest that PVX may recruit the WRKY transcription factor, which binds to the 5' NTR of viral genomic RNA and acts as a key regulator of viral infection.