• The Plant Pathology Journal
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• 제37권3호
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• pp.258-267
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• 2021

Asian pear (Pyrus pyrifolia) is a widely cultivated and commercially important fruit crop, which is occasionally subject to severe economic losses due to latent viral infections. Thus, the aim of the present study was to examine and provide a comprehensive overview of virus populations infecting a major pear cultivar ('Singo') in Korea. From June 2017 to October 2019, leaf samples (n = 110) of pear trees from 35 orchards in five major pear-producing regions were collected and subjected to RNA sequencing. Most virus-associated contigs matched the sequences of known viruses, including apple stem grooving virus (ASGV) and apple stem pitting virus (ASPV). However, some contigs matched the sequences of apple green crinkle-associated virus and cucumber mosaic virus. In addition, three complete or nearly complete genomes were constructed based on transcriptome data and subjected to phylogenetic analyses. Based on the number of virus-associated reads, ASGV and ASPV were identified as the dominant viruses of 'Singo.' The present study describes the virome of a major pear cultivar in Korea, and looks into the diversity of viral communities in this cultivar. This study can provide valuable information on the complexity of genetic variability of viruses infecting pear trees.

• 한국작물학회지
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• 제65권4호
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• pp.496-507
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• 2020

본 연구는 사과 바이러스 ASGV, ASPV 및 ApMV를 각각 정밀하게 진단하고자 SYBR Green I 및 TaqMan probe, 두 종류의 다른 chemical dye를 사용하여 quantitative real-time PCR 검정법을 개발하고자 하였다. 1. 사과 바이러스 ASGV, ASPV 및 ApMV의 국내분리주를 바탕으로 하여 cloning 및 in vitro transcription을 수행해 10배 희석단위 표준시료를 제작하였다. 각 바이러스에 대한 SYBR Green I용 프라이머와 TaqMan probe용 프라이머 및 프로브를 디자인하였다. 2. 상기 제작된 프라이머와 프로브를 이용해 표준시료를 대상으로 real-time PCR을 수행하여 각 바이러스의 증폭곡선과 검량선을 구할 수 있었다. Real-time PCR 결과, SYBR Green I기반의 검정법은 TaqMan probe기반의 검정법 못지 않은 결과를 보여주었으며, 적은 비용에 대량 검정이 요구되는 곳에 효과적으로 응용될 수 있을 것이다. 3. 현장평가를 본 실험에서 개발된 TaqMan probe기반의 real-time PCR검정법과 기존의 RT-PCR검정법과 비교분석하였다. 그 결과 real-time PCR 검정법은 singleplex 및 multiplex RT-PCR보다 더 민감하고 정확한 결과를 내어 RT-PCR로 검출할 수 없는 농도까지 검정할 수 있음을 입증하였다. 4. 본 실험에서 개발한 real-time PCR검정법은 검역현장과 같은 대량의 검사가 요구되는 곳에서는 SYBR Green I 기반의 검정법을, 바이러스 연구분야에서는 세밀한 정량이 가능한 TaqMan probe 방식의 검정법이 활용될 것으로 기대한다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 춘계학술대회
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• pp.54-54
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• 2019

Apple (Malus domestica) is one of the most economically important fruits in Korea. But virus infection has decreased sustainable production of apple and caused the serious problems such as yield loss and poor fruit quality. Virus or viroid infection including Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV), Apple mosaic virus (ApMV) and Apple scar skin viroid (ASSVd) has been also reported in Korea. In many cases, apple is infected with virus and viroid with no specific symptoms, the damage caused by the virus are unaware significantly. In our research, we tried to eliminate viruses in the rootstock for the disease-free seedlings of the apple dwarfing rootstock M.9 and M.26. The method of virus elimination was meristem culture, heat($37^{\circ}C$, 6weeks) treatment and chemistry($Ribavirin^{(R)}$) treatment. The analytical methods commonly used for the detection of virus is Enzyme-linked Immuno-Sorbent Assay(ELlSA) and Reverse Transcription-polymerase Chain Reaction(RT-PCR). RT-PCR method was more 30% sensitive than ELISA method. Efficiency of method eliminate virus appeared meristem method > heat treatment > chemistry treatment. The higher acquisition rate of disease-free seedlings is 30~40% on meristem treatment. In meristem treatment, the apple dwarfing rootstock M.9 gained infection ratio of ACLSV, ASPV and ASGV were 45%, 60% and 50% respectively. In the apple dwarfing rootstock M.26, infection ratio of ACLSV, ASPV and ASGV were 40%, 55%, 55%, respectively. Based on our results, it was found that most effective method of disease-free seedlings apple dwarfing rootstocks was by meristem treatment than heat method and chemistry treatment.

• Journal of Plant Biotechnology
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• 제49권3호
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• pp.222-229
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• 2022

사과는 국내 과수산업에서 가장 많이 재배되고 있는 과종이다. 하지만 apple mosaic virus (ApMV), apple stem grooving capillovirus (ASGV), apple stem pitting virus (ASPV), apple chlorotic leaf spot virus (ACLSV), apple scar skin viroid (ASSVd)와 같은 바이러스 및 바이로이드에 감염되면 과실의 수확량 감소 및 품질 저하를 야기시킨다. 본 연구에서는 국내 사과 농가에서 가장 많이 감염되어 있는 ASGV 바이러스를 제거하기 위한 효율적인 무병화 시스템을 확립하고자 하였다. ASGV에 감염된 폿트묘를 36℃, 38℃, 40℃가 유지되는 항온·항습장치에서 4주간 열처리를 수행하였으며, 신초 생장율과 바이러스 제거율을 조사하였다. 신초 생장률은 36℃ 처리구에서 가장 높았으며 신초의 중간부와 상단부는 바이러스가 제거되었으나 하단부는 바이러스가 제거되지 않았다. 38℃, 40℃ 처리구는 신초의 모든 구간에서 바이러스가 제거되지 않았으며, 40℃ 처리구는 신초의 생장 없이 열처리 3주 후 고사되었다. 36℃ 온도에서 열처리된 폿트묘의 경정을 절취하여 기외에서 접목하였으며 94%의 생존율과 20%의 바이러스 제거율을 보였다. 따라서 열처리 및 경정접목을 통해 무병묘 생산이 가능할 것으로 판단되었다.

• 농업과학연구
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• 제43권1호
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• pp.21-27
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• 2016

Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV), and Apple stem pitting virus (ASPV) have been known to induce top working disease causing economical damage in apple. Occurrences of these three viruses in pome fruit trees, including apple, have been reported around the world. The transmission of the three viruses was reported by grafting, and there was no report of transmission through mechanical contact, insect vector, or seed except some herbaceous hosts of ASGV. As RNA extraction methods for fruit trees, Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and multiplex RT-PCR techniques have been improved for reliability and stability, and low titer viruses that could not be detected in the past have become detectable. We studied the seed transmission ability of three apple viruses through apple seedling diagnosis using RT-PCR. Nineteen seeds obtained from commercially grown apple were germinated and two of the resulting plants were ASGV positive. Seven clones of the amplified ASGV coat protein (CP) genes of these isolates were sequenced. Overall sequence identities were 99.84% (nucleotide) and 99.76% (amino acid). Presence of a previously unreported single nucleotide and amino acid variation conserved in all of these clones suggests a possible association with seed transmission of these 'S' isolates. A phylogenetic tree constructed using ASGV CP nucleotide sequences showed that isolate S sequences were grouped with Korean, Chinese, Indian isolates from apple and Indian isolates from kiwi.