• Journal of Plant Biology
• /
• 제15권1호
• /
• pp.23-27
• /
• 1972

A total of 163 virus infected pepper plants(Capsicum annuum L.) collected from various pepper growing regions in Korea were investigated on the presence of tobacco mosaic virus (TMV), cucumber mosaic virus (CMV), potato virus X(PVX), potato virus Y(PVY) and alfalfa mosaic virus (AMV) by serological methods. Van Slogteren's microprecipitin test was applied for the testing of TMV, PVX and PVY from infected plants, and Ouchterlony agar double diffusion test was used for CMV and AMV. Results obtained are as follows: 1. TMV, CMV, PVX, PVY and AMV were found to occur on the pepper plants growing in Korea. 2. The prevalence of each of these viruses among the 163 pepper plants investigated was in the order of CMV: 93 plants(57.0%)>TMV: 91 plants (55.8%)>AMV: 58 plants (35.6%)>PVY: 40 plants (24.5%)> PVX:6 plants(3.7%). 3. Among the 163 plants investigated, 72 plants (44%) showed infection with one kind of virus and 91 plants (56%) showed mixed infection with more than two different viruses. In general, heavier damage of the plants was observed from mixed infection. 4. The results of serological identification of pepper viruses coincided with those results obtained by sap inoculation experiment conducted at the Horticultural Experiment Station along with present investigation. Thus the serological techniques applied in this experiment proved to be very reliable for the identification of TMV, CMV, PVX, PVY and AMV from pepper plants infected with these viruses.

• 미생물학회지
• /
• 제3권2호
• /
• pp.1-8
• /
• 1965

The potato viruses, as possible potato virus X(PVX), potato virus Y(PVY), potato virus S(PVS) are isolated from potato tuber, which collected from eleven areas (Table 1) in Korea. These viruses are isolated by single lesion isolation, Aphid transmission and inoculating methods through the many species of the different plants. The PVX is identified by host range, symptoms, physical properties, serological reaction and electron micrography. The other two viruses are identified by the first two methods mentioned above. The results of the above experiments are as follows. The total value of these viruses infection is 81%. The value of PVX infection is higher than the other two viruses. The properties of PVX are marked as local lesions on Comphrena globosa. The dilution end point is $10^{-6}$, the thermal inactivation point is $70^{\circ}C$ and the size of virus particles is around 13 x 60 $m\mu$.

• 한국응용곤충학회지
• /
• 제14권2호
• /
• pp.59-63
• /
• 1975

우리나라 잎담배에 발생하고 있는 바이러스의 종류와 감염상을 조사하기 위하여 전주지방에서 Burley종 (Burley-21) 20개체, 부천연초시험장 포장에시 황색종(Hicks) 20개체를 병징별로 채집하여 혈청학적인 방법에 의해 실시했다. 검정대상 바이러스는 Tobacco mosaic virus(TMV), Cucumber mosaic virus(CMV), Alfalfa mosaic virus (AMV), Potato virus X(PVX) 및 Potato virus Y(PVY)등 5종으로 TMV, AMV, CMV는 한천내확산법으로 PVX, PVY는 미량침강법으로 실시하여 다음과 같은 결과를 얻었다. 1. 공시이병개체중에서 TMV, CMV, AMV, P X 및 PVY 등 5종의 바이러스가 검출되었다. 2. 조사된 40개체중 각 바이러스의 감염율은 AMV $67.5\%$, CMV $60.0\%$, TMV$47.5\%$PVY$17.5\%$, PVX$10.0\%$의 순이었다. 3. Burley종에서는 TMV감염율이 $15.0\%$로 황색종의 $80.0\%$보다 현저히 낮았다. 4. 조사된 각 개체는 단독감염으로부터 2-4종의 바이러스로 다양하게 혼합감염을 나타냈으며 이 중 단독감염이 $37.5\%$, 혼합감염이 $62.5\%$이었다.

• 한국식물병리학회:학술대회논문집
• /
• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
• /
• pp.74.1-74
• /
• 2003

Differential display (DD) of mRNA is a technique in which mRNA species expressed by a cell population are reverse transcribed and then amplified by many separate polymerase chain reactions (PCR). Using DD-RT-PCR we obtained many genes that expressed differentially in healthy and PVX-infected Nicotiana benthamima, using total RNAs extracted from healthy and PVX-infected N. benthamiana plants. Three hundred and twenty-five DNA fragments isolated from DD-RT-PCR were cloned and sequenced for further characterization. Several host genes including SKPI-like protein, heat shock transcription factor and Avr9/Cf-9 rapidly elicited protein were selected to obtain full-length open reading frame and to characterize their potential involvement in virus disease development and/or host's defense against virus infection employing PVX-based expression vector. Transcrips from wild-type and clones containing each selected gene were inoculated onto N. benthamiana Levels of virus replication were confirmedby RT-PCR and RNA blot analysis, Expression profiles and potential role(s) of selected genes upon PVX infection will be discussed.

• The Plant Pathology Journal
• /
• 제22권2호
• /
• pp.139-146
• /
• 2006

The 3' region of Potato virus X (PVX) has the 74 nt 3'-nontranslated region (NTR) that is conserved among all potexviruses and contains several cis-acting elements for minus-strand and plus-strand RNA accumulation. Three stem-loop structures (SL1-SL3), especially formation of SL3 and U-rich sequence of SL2, and near upstream elements in the 3' NTR were previously demonstrated as important cis-acting elements. To Investigate the binding of these cis-acting elements within 3' end with host protein, we used the electrophoretic mobility shift assays (EMSA) and UV-cross linking analysis. The EMSA with cellular extracts from tobacco and RNA transcripts corresponding to the 150 nt of the 3' end of PVX RNA showed that the 3' end of PVX formed complexes with cellular proteins. The specificity of protein binding was confirmed through competition assay by using with 50-fold excess of specific and non-specific probes. We also conducted EMSA with RNAs containing various mutants on those cis-acting elements (${\Delta}10$10, SL3B, SL2A and ${\Delta}21$; J Mol Biol 326, 701-720) required for efficient PVX RNA accumulation. These analyses supported that these cis-acting elements are required for interaction with host protein(s). UV-cross linking analysis revealed that at least three major host proteins of about 28, 32, and 42 kDa in mass bound to these cis-elements. These results indicate that cis-acting elements from 3' end which are important for minus and plus-strand RNA accumulation are also required for host protein binding.

• The Plant Pathology Journal
• /
• 제24권3호
• /
• pp.289-295
• /
• 2008

RNA-RNA interactions and the dynamic RNA conformations are important regulators in virus replication in several RNA virus systems and may also involved in the regulation of many important virus life cycle phases, including translation, replication, assembly, and switches in these important stages. The 5' non-translated region of Potato virus X(PVX) contains multiple cis-acting elements that facilitate various viral processes. It has previously been proposed that RNA-RNA interactions between various RNA elements present in PVX RNA genome are required for PVX RNA accumulation(Hu et al., 2007; Kim and Hemenway, 1999). This model was based on the potential base-pairing between conserved sequence elements at the upstream of subgenomic RNAs(sgRNAs) and at the 5' and 3' end of RNA genome. We now provide more evidence that RNA-RNA base-pairing between elements present at the 5' end and upstream of each sgRNA is required for efficient replication of genomic and subgenomic plus-strand RNA accumulation. Site-directed mutations introduced at the 5' end of plus-strand RNA replication defective mutant(${\Delta}12$) increasing base-pairing possibility with conserved sequence elements located upstream of each sgRNAs restored genomic and subgenomic plus-strand RNA accumulation and caused symptom development in inoculated Nicotiana benthamiana plants. Serial passage of a deletion mutant(${\Delta}8$) caused more severe symptoms and restored wild type sequences and thus retained possible RNA-RNA base-pairing. Altogether, these results indicate that the RNA element located at the 5' end of PVX genome involved in RNA-RNA interactions and play a key role in high-level accumulation of plus-strand RNA in vivo.

• Journal of Plant Biology
• /
• 제5권3호
• /
• pp.30-36
• /
• 1962

The mechanism of synthesis of the toacco mosaic virus(TMV) and the potato virus X(PVX) was investigated using the methods of ultraviolet light irradiation and serological analysis. In vitro irradiation of UV on the infected tobacco juice for 10 minutes caused the infectivity of TMV and PVX to decrease markedly on their respective local lesion indicator hosts, Nicotiana glutinosa L. and Gomphrena globosa L., indicating that UV destroys directly the infectivity of the virus particles. Ten minutes after the UV was irradiated on the leaves of the two indicator hosts before inoculation, the infectivity of TMV decreased as it was irradiated in vitro, whereas that of PVX increased by 26% as compared with the unirradiated control. When the two viruses were mix-inoculated in the common host of tobacco and the synthetic products were analyzed by serological methods for a two week infection period, it was found that both viruses were multiplying more rapidly and abundantly than they were singly inoculated into the same host species. Titers from mixed series were often two times as high as those of singly inoculated series. A mechanism of competition in the synthesis between the mixed viruses in the common host is postulated.

• Journal of Plant Biotechnology
• /
• 제5권2호
• /
• pp.101-105
• /
• 2003

To eradicate several viruses such as PVX, PVY, and PLRV which often cause considerable damages to the growth and yields of potatoes, several stems including shoot tips were excised from the potato plants grown for 50 days and electric shock was treated. Shoot tips excised from electric-shocked stems were transferred into the medium supplemented with antiviral compound, ribavirin to examine the combinatorial effect. When treated only with 20 mg/L ribavirin, PVX concentration in the regenerated plant-lets was slowly decreased as repeating sub-culture and finally, it took 32 weeks to reach completely PVX-free stock. With an electric shock treatment (10 mA electric current), all the replicates became free from PVY. However, PLRV was not completely eradicated from 94P70-4 and 93P29-3 lines even by treating with 10 mA electric shock. In this case, both electric shock and antiviral compound treatments in axillary buds from the stem segment were successful in eradicating viral contamination.

• 미생물과산업
• /
• 제17권3호
• /
• pp.86-88
• /
• 1991

한국의 중부지역을 중심한 11개 지역으로부터 수집한 자료를 재료로 하여 감자바이러스를 분석한 결과 potato virus X, potato virus Y, potato virus S의 3종류의 감자바이러스 계통이 우리나라에 분포하고 있음을 알았다. 이들의 민합감심 비율은 81%를 나타냈으며 강원도 지방이 가장 적었다. 기중 pvx의 제성질을 조사한 결과 dilution end point는 $10^{-6}$ , thermal inactivation point는 7$0^{\circ}C$를 나타냈다. 입자의 크기는 550~650ml사이였다. 기중 600ml이 80%를 갖는 PVX계통이였다.

• The Plant Pathology Journal
• /
• 제31권3호
• /
• pp.219-225
• /
• 2015

The primary step for efficient control of viral diseases is the development of simple, rapid, and sensitive virus detection. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has been used to detect viral RNA molecules because of its simplicity and high sensitivity for a number of viruses. RT-LAMP for the detection of Potato virus X (PVX) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to demonstrate its advantages over RT-PCR. RT-LAMP reactions were conducted with or without a set of loop primers since one out of six primers showed PVX specificity. Based on real-time monitoring, RT-LAMP detected PVX around 30 min, compared to 120 min for RT-PCR. By adding a fluorescent reagent during the reaction, the extra step of visualization by gel electrophoresis was not necessary. RT-LAMP was conducted using simple inexpensive instruments and a regular incubator to evaluate whether RNA could be amplified at a constant temperature instead of using an expensive thermal cycler. This study shows the potential of RT-LAMP for the diagnosis of viral diseases and PVX epidemiology because of its simplicity and rapidness compared to RT-PCR.