• 한국식물병리학회지
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• 제11권2호
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• pp.101-106
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• 1995

Deep blue fluorescence of resorcin blue-stained callose was observed only in the potato leafroll virus (PLRV)-infected potato plants, but not in other potato viruses investigated. The plant sections stained with aniline blue showed non-specific fluorescence regardless of PLRV infection, which means that aniline blue is not suitable for the staining of callose induces by PLRV infection. The fluorescence of resorcin blue-stained callose was more easily detectable than autofluorescence by a direct fluorescence detection method because of its deep blue color. The lateral branch of lower leaves was turned out to be the best material for fluorescence observation of all plant parts tested. In comparison of diagnostic efficacy of this technique to enzyme-linked immunosorbent assay (ELISA), PLRV infected potato plants showed corresponding increment of the fluorescence of resorcin blue stained callose as absorption values in ELISA increased. In the future, the criteria measuring the fluorescence objectively are thought to be determined for the practical application to the diagnosis of potato leafroll disease.

• 한국식물병리학회지
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• 제14권6호
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• pp.636-641
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• 1998

Digoxigenin (DIG) was used to prepare nucleic acid probe for the detection of RNA of potato leafroll virus (PLRV) in the potato leaf extracts. The 0.6 kb coat protein (CP) gene cDNA of PLRV in plasmid pSPT 18 vector was labeled with digoxigenin by in vitro run-off transcription and then used for cRNA probe. In the several buffers tested for increase the total RNA extraction efficiency AMES buffer was the most suitable for this detection method. The RNA extracts from potato leaves shown symptoms of PLRV were dot blotted onto nylon membrane and hybridized with labeled RNA probes. After hybridization, labeled RNA bound to PLRV RNA on membrane was detected with anti-digoxigenin alkaline phosphatase. 5-bromo-4-chloro-3-indolyl-phosphate/nitroblue tetrazolium (NBT) salt and CSPD were used as substrate for colorimetric and film exposure detection, respectively. These detection methods were very sensitive allowing for detection of 1/32 diluted total RNA extract from 100 mg leaf tissue.

• The Plant Pathology Journal
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• 제32권4호
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• pp.321-328
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• 2016

We examined the effects of temperature on acquisition of Potato virus Y-O (PVY-O), Potato virus A (PVA), and Potato leafroll virus (PLRV) by Myzus persicae by performing transmission tests with aphids that acquired each virus at different temperatures. Infection by PVY-O/PVA and PLRV increased with increasing plant temperature in Nicotiana benthamiana and Physalis floridana, respectively, after being transmitted by aphids that acquired them within a temperature range of $10-20^{\circ}C$. However, infection rates subsequently decreased. Direct qRT-PCR of RNA extracted from a single aphid showed that PLRV infection increased in the $10-20^{\circ}C$ range, but this trend also declined shortly thereafter. We examined the effect of temperature on establishment of virus infection. The greatest number of plants became infected when N. benthamiana was held at $20^{\circ}C$ after inoculation with PVY-O or PVA. The largest number of P. floridana plants became infected with PLRV when the plants were maintained at $25^{\circ}C$. PLRV levels were highest in P. floridana kept at $20-25^{\circ}C$. These results indicate that the optimum temperatures for proliferation of PVY-O/PVA and PLRV differed. Western blot analysis showed that accumulations of PVY-O and PVA coat proteins (CPs) were lower at $10^{\circ}C$ or $15^{\circ}C$ than at $20^{\circ}C$ during early infection. However, accumulation increased over time. At $25^{\circ}C$ or $30^{\circ}C$, the CPs of both viruses accumulated during early infection but disappeared as time passed. Our results suggest that symptom attenuation and reduction of PVY-O and PVA CP accumulation at higher temperatures appear to be attributable to increased RNA silencing.

• The Plant Pathology Journal
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• 제28권3호
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• pp.248-258
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• 2012

Potato (Solanum tuberosum) is one of the most important food crops worldwide and yields of potato can be affected by virus infection. While more than 40 viruses have been found in potato, only nine viruses (potato leafroll virus, potato viruses A, M, S, V, X and Y, potato moptop virus and tobacco rattle virus) and one viroid (potato spindle tuber viroid) have a significant economic impact on potato, worldwide. This review describes the geographical distribution of the most important viruses infecting potato and the genes for resistance or tolerance that have been identified against these various infectious agents. In some cases such resistance genes have been found only in other Solanum species. Few genes for resistance to the vectors of these viruses have been obtained and even fewer have been deployed successfully. However, transgenic resistance in potato has been achieved against seven of these disease agents.

• The Plant Pathology Journal
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• 제27권4호
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• pp.385-389
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• 2011

A new reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the Potato leafroll virus (PLRV) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to address its advantages over RTPCR. RT-LAMP primers were designed from the open reading frame 3 (ORF3) sequence of PLRV. The RT-LAMP reactions were conducted without or with a set of loop primers. By real-time monitoring using Turbimeter, the RT-LAMP (with loop primers) detects PLRV in less than 30 min, compared to 120 min of RT-PCR. By adding fluorescent reagent during the reaction, final products of the RT-LAMP were fluorescently visualized under UV light or could be differentiated by naked-eye inspection under normal light. The RT-LAMP was extremely sensitive, about 2000-fold more sensitive than RT-PCR. This study presents great potential of the RT-LAMP for diagnosis and PLRV epidemiology because RT-LAMP method is speedy, sensitive, inexpensive, and convenient.

• 식물병연구
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• 제9권1호
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• pp.1-9
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• 2003

The occurrence of potato(Sotanum tuberosum) viral diseases caused by Potato virus X(PVX), Potato virus Y (PVY), Potato leafroll virus(PLRV), Potato vims S(PVS), Potato virus M(PVM), Potato virus A(PVA), Potato virus T(PVT), Alfalfa mosic virus(AIMV), Tobacco mosic virus(TMV), Potato mop top virus(PMTV) Tobacco rattle virus(TRV) and Potato spindle tuber viroid(PSTVd), potato witches' broom phytoplasma, have been identified so far in Korea. Major viral diseases such as PVX, PVY and PLRV had been studied more deeply, however, the others are just identified and only partially characterized since the first study on the relation between PVX nucleic acid and virus protein by Kim in 1961. The most studies on potato viral diseases are mainly focused on the problems of seed potato production. The National Alpine Agricultural Experiment Station(NAAES), since it began its activities in 1961, has given special attention to this problem by doing studies to identify, characterize and control potato virus diseases. This effort resulted in the development of new potato virus detection methods as a basis for elaborating new method of control, such as the production of seed potato free of virus and the selection of new virus-resistant transgenic potatoes. The further studies of potato viral diseases required would be fallowings: the continuous monitoring for the occurrence of identified or not identified potato viruses in Korea, the isolation of resistant viral genes, the development of control method for the non-persistently transmitted viruses like PVY, special vectors such as nematode and fungus transmitted viruses, TRV and PMTV and the development of control methods against potato viral diseases by viral cross protection, therapy, transgenic plant, and the use of the agents or molecules, such as virus inhibitors and antiviral proteins, etc., blocking viral replication.

• 한국잠사학회:학술대회논문집
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• 한국잠사학회 2003년도 International Symposium of Silkworm/Insect Biotechnology and Annual Meeting of Korea Society of Sericultural Science
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• pp.116-117
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• 2003

Aphidicidal activity of Bacillus thurigiensis crystal proteins was recently reported. However, relatively higher dose of crystal protein was needed to kill aphids. In this study, we intended to improve the aphidicidal activity of crystal protein by fusion with coat protein (CP) of potato leafroll virus (PLRV) which is transmitted by aphids. (omitted)

• The Plant Pathology Journal
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• 제33권1호
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• pp.101-101
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• 2017

• 한국작물학회지
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• 제59권4호
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• pp.498-504
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• 2014

Potato virus Y (PVY) and potato leafroll virus (PLRV) are among the most damaging potato viruses and prevalent in most potato growing areas. In this study, cryopreservation was used to eradicate PVY and PLRV using two cryogenic methods. Potato shoot tips proliferated in vitro were cryopreserved through droplet-vitrification and encapsulation-vitrification using plant vitrification solution 2 (PVS2; 30% glycerol + 15% dimethyl sulfoxide + 15.0% ethylene glycol + 13.7% sucrose) and modified PVS2. Both cryogenic procedures produced similar rates of survival and regrowth, which were lower than those from shoot tip culture alone. The health status of plantlets regenerated from shoot tip culture alone and cryopreservation was checked by reverse transcription-polymerase chain reaction. The frequency of virus-free plants regenerated directly from highly proliferating shoot tips reached 42.3% and 48.6% for PVY and PLRV, respectively. In comparison, the frequency of PVY and PLRV eradication after cryopreservation was 91.3~99.7% following shoot-tip culture. The highest cryopreserved shoot tip regeneration rate was observed when shoot tips were 1.0~1.5 mm in length, but virus eradication rates were very similar (96.4~99.7%), regardless of shoot tip size. This efficient cryotherapy protocol developed to eliminate viruses can also be used to prepare potato material for safe long-term preservation and the production of virus-free plants.

• 한국식물병리학회지
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• 제12권3호
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• pp.358-362
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• 1996

감자 잎말림 바이러스를 검정하기 위하여 ELISA 및 전자현미경에 의해 바이러스 감염이 확인된 기내 배양중인 감장의 줄기로부터 RT-PCR 분석을 수행하였다. 분리된 총 RNA들로부터 바이러스 cDNA를 합성하고 감자 잎말림 바이러스 외피단백질의 일부인 465bp를 특이하게 증폭하도록 고안한 두 primer를 사용하여 PCR 반응을 하였다. 증폭된 465pb의 DNA 절편의 염기서열을 분석한 결과 역시 감자 잎말림 바이러스임을 확인하였다. 바이러스 검정에 있어서 EL-ISA 방법과 RT-PCR 방법간의 민감도를 조사한 결과 RT-PCR 방법간의 민감도를 조사한 결과 RT-PCR 방법이 ELISA 방법보다 감자 잎말림 바이러스검정에 있어서보다 정확한 방법인 것으로 사료된다.