• The Plant Pathology Journal
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• 제20권2호
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• pp.127-130
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• 2004

Grapevine leaf roll-associated virus 3 (GLRaV-3) is one of the most important viral diseases of grapevine in the world. In this study, GLRaV-3 Ampelovirus was identi-fied from grapevines in Korea by analyzing viral coat protein size, nucleotide, and amino acid sequences. The molecular weight of viral coat protein from virus-infected in vitro plantlets was determined by western blot using a commercial GLRaV-3 polyclonal antibody. Western blot analysis showed a coat protein of about 43 kDa. RT-PCR product of about 942 bp which encoded the coat protein (CP) gene was amplified with specific primers. When the viruses existed at low titers in the host plant, the dsRNA had very specific template in RT- PCR amplification of fruit tree viruses. Especially, small-scale dsRNA extraction method was very reliable and rapid. Sequence analysis revealed that the CP of the GLRaV-3 Ko consisted of 942 bp nucleotide, which encoded 314 amino acid residues. The CP gene of GLRaV-3 Ko had 98.9% nucleotide sequence and 98.7% amino acid sequence identities with earlier reported GLRaV-3. This is the first report on molecular assay of GLRaV-3 Ampelovirus identified from Korea. The GLRaV-3 Ko CP clone would be very useful for breeding of virus resistant grapevines.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.138.2-139
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• 2003

Grapevine leafroll-associated 1 virus (GLRaV-1) and Grapevine leafroll-associated 3 virus (GLRaV-3), member of the genus Ampelovirus, are important viral disease of grapevine in the world. these viruses transmitted only dicotyledonous host by vectors such as mealybugs and there is no suitable herbaceous host for virus. The diseased leaves turn yellowish or reddish depending on cultivars and viruses. Viruses are existed at low concentration and ununiformly distribution in grapevine. Using small-scale double-stranded RNA (dsRNA) extraction method, reverse transcription and polymerase chain reaction (RT-PCR) product of 1Kb long which encoded of coat protein (CP) gene for both viruses was successfully amplified with a specific primers. The RT-PCR product was cloned into the plasmid vector and its nucleotide sequences were determined from selected recombinant cDNA clones. Sequence analysis revealed that the CP of GLRaV-1 consisted of 969 nucleotide, which encoded 323 amino acid residues and CP of GLRaV-3 consisted of 942 nucleotide, which encoded 314 amino acid residues. The CP of GLRaV-1 and GLRaV-3 has 93.8％ and 98.7％ amino acid sequence identities, respectively.

• Journal of Plant Biotechnology
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• 제30권2호
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• pp.155-160
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• 2003

Grapevine leafroll-associated virus 3(GLRaV-3) is one of the most severe pathogens for viral diseases found in Korea. This study was conducted to establish the virus-free stock production system for the virus disease control. The effects of thermotherapy, merestem culture and chemotheratpy to eliminate the GLRaV-3 in gratevines were tested. Thermotherapy at 37$\pm$2$^{\circ}C$ for 6∼8 weeks combined with 0.5∼1.0mm size of meristem culture method was the most effective for virus elimination. Thermotherapy alone was not effective. In chemotheratpy, DHT and Amantadine (20, 40mg/L) treatment in medium was more effective than Ribavirin to eliminate the GLRaV-3 in grapevine. However, Ribavirin spraying to potted was not available for virus elimination. Therefore, virus-free stock production system using the thermotherapy combined with shoot apical meristem culture was the most effective in grapevine.

• 식물조직배양학회지
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• 제28권6호
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• pp.335-339
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• 2001

우리나라 포도원에서 가장 많이 발생하여 피해를 주고 있는 grapevine leafroll-associated 3 Closterovirus (GLRaV-3)를 대상으로 기내 배양묘를 이용한 바이러스 순화 및 진단효율성을 검토하였다. 바이러스 감염주의 절간을 기내배양하여 증식시킨 기내 배양묘를 1개월 간격으로 계대배양하면서 배양묘를 ELISA 검정한 결과 고농도의 바이러스가 검출되었으며 배양묘의 부위별로 잎, 줄기 및 줄기 유래의 callus 조직에서 모두 고농도로 검출되었다. 또한 포장에서 재배되고 있는 포도나무의 잎이나 엽병조직에 비해 기내 배양묘의 조직을 사용하였을 때 높은 농도의 정제 바이러스를 얻을 수 있었으며, 전자현미경에 의한 dip 검경에서도 사상형 바이러스 입자가 관찰되었다. RT-PCR 진단에서는 기내 배양묘 조직을 사용하였을 때 포도 유엽조직과 엽병+중륵조직에 비해 검출효율이 높았다. 기내 배양묘의 잎조직을 이용하여 ELISA와 RT-PCR 검정감도를 비교한 결과 ELISA에 비해 RT-PCR 검정이 약 1,000배 감도가 높았다.

• 식물병연구
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• 제30권2호
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• pp.176-180
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• 2024

본 연구에서는 2021년 충청북도 및 경상북도에 위치한 7개 지역의 총 29개 포도 과원에서 재배되고 있는 포도 5개 품종에서 총 217의 시료를 채집하였다. 바이러스의 진단을 위해서 5종 바이러스에 대한 종 특이프라이머를 이용하여 중합효소연쇄반응하였다. 진단 결과 grapevine fleck virus (GFkV)는 126개 시료(58.06%)에서, grapevine leafroll-associated virus (GLRaV)-3는 112개 시료(51.61%)에서, GLRaV-1는 4개 시료(1.84%)에서 확인되었다. 특히 샤인머스캣과 알렉산드리아 품종에서의 감염률이 높았으며, 지역별로는 경북 지역이 높은 감염률을 보였다. 그러나 grapevine red blotch virus (GRBV)와 grapevine berry inner necrosis virus (GINV)는 확인되지 않았다.

• The Plant Pathology Journal
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• 제18권5호
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• pp.244-250
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• 2002

Grapevine leafroll-associated 3 virus (GLRaV-3) and Grapevine fanleaf virus (GFLV) are important viral diseases of grapevine in the world. In this study, the most reliable woody indicator plants were selected for virus indexing. Two grapevines, LN33 (Couderc 1613x vitis berlandieri) and Vitis riparia Gloire, were selected for CLRaV-3 and CFLV graft indexing, respectively. The specific characteristics of Closterovirus isolated from grapevines cultivated in Korea were identified. filamentous virus-like particles only existed in the phloem parenchyma cell. In particular, the vesiculation of mitochondria was observed. This mitochondrial vesicu-lation was considered to be one of the most reliable cytopathic features of Closterovirus. During observation of GFLV-infected Chenopodium quinoa sections, virus-like particles arranged consistently were found forming several layers in cytoplasm. Moreover, virus-like particles in tubules were observed and were associated with plasmodesmata in cytoplasm. This is the first report on cytopathological characteristics of Closterovirus and Nepovirus identified from grapevines in Korea.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 1999년도 임시총회 및 추계 학술발표회
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• pp.83.1-83
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• 1999

• The Plant Pathology Journal
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• 제33권6호
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• pp.561-571
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• 2017

Prospecting of local grapevine (Vitis vinifera L.) germplasm revealed that Tunisia possesses a rich patrimony which presents diversified organoleptic characteristics. However, viral diseases seriously affect all local grapevine cultivars which risk a complete extinction. Sanitation programs need to be established to preserve and exploit, as a gene pool, the Tunisian vineyards areas. The presence of the Grapevine leafroll associated virus-3 (GLRaV-3), Grapevine stem pitting associated virus (GRSPaV) and Grapevine virus A (GVA), were confirmed in a Tunisian grapevine cultivar using serological and molecular analyses. The association between GRSPaV and GVA viruses induces more rugose wood symptoms and damages. For this reason the cleansing of the infected cultivar is highly advisable. Direct and recurrent somatic embryos of cv. 'Hencha' were successfully induced from filament, when cultured on $Ch{\acute{e}}e$and Pool (1987). based-medium, enriched with $2mg1^{-1}$ of 2,4-dichlorophenoxyacetic acid and $2.5mg1^{-1}$ of Thidiazuron, after 36 weeks of culture. After six months of acclimatization, RT-PCR carried on 50 somaplants confirmed the absence of GVA, GRSPaV as well as GLRaV-3 viruses in all somaplants. Ampelographic analysis, based on eight OIV descriptors, was carried out on two years acclimated somaplants, compared to the mother plant. Results demonstrated that the shape and contours of 46 somaclones leaves are identical to mother plant leaves and four phenotypically off-type plants were observed. The healthy state of 100% 'Hencha' somaclones and the high percentage of phenotypically true-to-type plants demonstrate that somatic embryogenesis is a promising technique to adopt for grapevine viruses elimination.