• The Plant Pathology Journal
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• 제32권5호
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• pp.441-451
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• 2016

Deep sequencing has generated 52 contigs derived from five viruses; Apple chlorotic leaf spot virus (ACLSV), Apple stem grooving virus (ASGV), Apple stem pitting virus (ASPV), Apple green crinkle associated virus (AGCaV), and Apricot latent virus (ApLV) were identified from eight apple samples showing small leaves and/or growth retardation. Nucleotide (nt) sequence identity of the assembled contigs was from 68% to 99% compared to the reference sequences of the five respective viral genomes. Sequences of ASPV and ASGV were the most abundantly represented by the 52 contigs assembled. The presence of the five viruses in the samples was confirmed by RT-PCR using specific primers based on the sequences of each assembled contig. All five viruses were detected in three of the samples, whereas all samples had mixed infections with at least two viruses. The most frequently detected virus was ASPV, followed by ASGV, ApLV, ACLSV, and AGCaV which were withal found in mixed infections in the tested samples. AGCaV was identified in assembled contigs ID 1012480 and 93549, which showed 82% and 78% nt sequence identity with ORF1 of AGCaV isolate Aurora-1. ApLV was identified in three assembled contigs, ID 65587, 1802365, and 116777, which showed 77%, 78%, and 76% nt sequence identity respectively with ORF1 of ApLV isolate LA2. Deep sequencing assay was shown to be a valuable and powerful tool for detection and identification of known and unknown virome in infected apple trees, here identifying ApLV and AGCaV in commercial orchards in Korea for the first time.

• 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.

• The Plant Pathology Journal
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• 제38권3호
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• pp.220-228
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• 2022

Pollen is a vector for viral transmission. Pollenmediated viruses cause serious economic losses in the fruit industry. Despite the commercial importance of pollen-associated viruses, the diversity of such viruses is yet to be fully explored. In this study, we performed metatranscriptomic analyses using RNA sequencing to investigate the viral diversity in imported apple and kiwifruit pollen. We identified 665 virus-associated contigs, which corresponded to four different virus species. We identified one virus, the apple stem grooving virus, from pear pollen and three viruses, including citrus leaf blotch virus, cucumber mosaic virus, and lychnis mottle virus in kiwifruit pollen. The assembled viral genome sequences were analyzed to determine phylogenetic relationships. These findings will expand our knowledge of the virosphere in fruit pollen and lead to appropriate management of international pollen trade. However, the pathogenic mechanisms of pollen-associated viruses in fruit trees should be further investigated.

• 식물병연구
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• 제26권2호
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• pp.95-102
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• 2020

사과 바이러스 4종(Apple chlorotic leaf spot virus [ACLSV], Apple stem pitting virus [ASPV], Apple stem grooving virus [ASGV], Apple mosaic virus [ApMV]), 바이로이드 1종(Apple scar skin viroid [ASSVd])을 대상으로 국내 감염률을 조사한 결과, 감염률은 97.3%로 대부분의 사과나무가 바이러스 및 바이로이드에 감염되어 있는 것으로 조사되었다. 지역별로는 정선 98.8%, 단양 100%, 예산 100%, 장수 89.1%, 무주 98.1%였으며, 바이러스 및 바이로이드 각각의 감염률은 ASGV 93.4%, ASPV 85.7%, ACLSV 59.0%, ASSVd 6.7%, ApMV 3.6% 순으로 ASGV의 감염률이 가장 높았고 ApMV의 감염률이 가장 낮은 것으로 조사되었다. 또한, 바이러스 및 바이로이드 2종 이상 복합 감염비율은 84.8%로 단 1종만 감염된 비율인 12.4%와 비교하여 약 7배 가까이 되는 것으로 조사되었다.

• 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.

• 식물병연구
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• 제25권1호
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• pp.43-47
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• 2019

Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV), and Apple scar skin viroid (ASSVd) are economically important viruses that infect pear tree species worldwide. To evaluate the prevalence of these viruses in Korea, we investigated infection degree of three viruses and one viroid for the commercial nursery trees of the pear cultivars, Niitaka, Chuwhang, Wonwhang, and Whasan in 2017 and 2018. The results showed that the infection ratio of ACLSV, ASPV, ASGV, and ASSVd for the scion of pear cultivar Niitaka was 10%, 45%, 77%, and 50%, respectively. From the scion of pear cultivar Chuwhang, infection ratios of ASPV, ASGV, and ASSVd were found to be 70%, 50%, and 60%, respectively. From the scion of pear cultivar Whasan, infection ratios of ACLSV, ASPV, ASGV and ASSVd were found to be 40%, 60%, 93%, and 20%, respectively. From the root stock of pear cultivar Wonwhang, infection ratios of ACLSV, ASPV, ASGV, and ASSVd showed 28%, 57%, 100%, and 14%, respectively. ASGV had the highest recorded infection rate, and ACLSV was characterized by the lowest infection rate. The mixed infection ratio of Niitaka, Chuwhang, Whasan, and Wonwhang was 45%, 60%, 70%, and 85%, respectively.

• The Plant Pathology Journal
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• 제27권1호
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• pp.44-52
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• 2011

Field surveys were conducted in 45 stone fruit orchards in seven districts of Isparta Province located in western Mediterranean region of Turkey important for stone fruit production. Leaf samples were collected from 175 trees showing virus-like symptoms. These samples were first tested by ELISA for five different RNA viruses including Apple mosaic ilarvirus (ApMV), Prunus necrotic ringspot ilarvirus (PNRSV), Prune dwarf ilarvirus (PDV), Plum pox potyvirus (PPV), Apple chlorotic leafspot trichovirus (ACLSV). While no ApMV and PPV infection was found, 46, 24 and 16 samples were tested positive for PDV, ACLSV and PNRSV, respectively, in ELISA showing about 45% of symptomatic trees in the region were infected with at least one of these viruses. In addition, it was found that nine sweet cherry trees were mixed infected with two or three of these viruses and PDV with an infection rate of 26.3% was the most widespread virus in symptomatic trees in western Mediterranean region. Thirty samples were selected and tested by a multiplex RT-PCR (mRT-PCR) for simultaneous detection of these viruses. While PPV was not detected, more than half of the tested 20 samples were individually or mixed infected with ApMV, ACLSV, PNRSV and PDV. The mRT-PCR results were confirmed by detection of these viruses individually in some of the field samples using RT-PCR with primes specific to each virus. Comparison of ELSA and mRT-PCR results of 30 samples showed that numbers of infected and mixed infected samples as well as infection and mixed infection rates were significantly higher in RT-PCR (20 and 66.7%) than in ELISA (14 and 46.7%). The results confirm that mRT-PCR is more sensitive than ELISA.

• 식물병연구
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• 제19권4호
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• pp.288-293
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• 2013

국내외적으로 ACLSV, ASGV, ApMV, ASSVd와 같은 바이러스 및 바이로이드 병의 발생으로 사과 과실의 생산량 감소와 기형적인 외형 등 많은 문제점들이 보고되었다. 하지만 사과 바이러스의 감염에 대한 방제 대책은 거의 알려진 바가 없는 실정이다. 따라서 본 논문에서는 사과 신품종인 '단홍', '홍안', '새나라', '썸머드림'을 분양하기에 앞서 바이러스 무병묘를 생산하는 시스템을 확립하고자 하였다. $37^{\circ}C$가 유지되는 항온 항습장치에서 4주간 열처리를 하였으며 기내에서 경정 배양을 하였다. 열처리된 각각의 사과 신품종들은 바이러스 진단 프라이머를 통해 RT-PCR을 수행하여 바이러스 진단을 수행하였다. 결과적으로 '단홍'은 28%의 바이러스 무병묘를 확보할 수 있었으며 '홍안'은 16%, '새나라'와 '썸머드림'은 12%의 확률로 바이러스 무병 사과를 확보할 수 있었다. 본 연구결과는 열처리 및 경정배양을 통해 사과 신품종에서 바이러스 무병묘 생산 시스템 구축이 가능함을 보여주었다.

• 식물병연구
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• 제23권4호
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• pp.355-362
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• 2017

국내에서 유통되고 있는 사과나무 묘목의 바이러스 감염 실태를 파악하기 위하여 사과 '홍로'와 '후지' 및 '후지' 아조변이 품종을 구입하여 5종류의 바이러스에 대한 감염 여부를 검정하였다. 그 결과, '홍로' 품종의 접수 부위는 ACLSV, ASPV 및 ASGV에 각각 100%, 81.3% 및 100%의 감염률을 보였고 ApMV와 ASSVd에는 전혀 감염되지 않았다. '홍로' 품종의 대목 부위에 대하여, ACLSV, ASPV 및 ASGV의 감염률은 각각 87.5%, 81.3% 및 100%이었고 ApMV와 ASSVd의 감염률은 각각 12.5% 및 6.3%이었다. '후지' 품종 및 '후지' 아조변이 품종의 접수 부위는 ACLSV, ASPV 및 ASGV에 각각 86.7%, 86.7% 및 100%의 감염률을 보였고 ApMV와 ASSVd에는 전혀 감염되지 않았다. 대목 부위는 ACLSV, ASPV 및 ASGV에 각각 86.7%, 93.3% 및 93.3%의 감염률을 보였고 ApMV와 ASSVd에 대한 감염률은 각각 12.5%와 6.3%이었다.

• Journal of Plant Biotechnology
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• 제46권3호
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• pp.228-235
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• 2019

사과(Malus pumila)는 국내에서 가장 경제적으로 중요한 과수 중의 하나이다. 하지만 사과 바이러스 감염은 생산량을 감소시키고 수확량 손실과 과일 품질 저하와 같은 심각한 문제를 야기한다. 국내에 감염된 사과 바이러스 및 비로이드 종류는 Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV), Apple mosaic virus (ApMV)와 Apple scar skin viroid (ASSVd) 등이 알려져 있다. 사과는 바이러스나 비로이드에 감염되어 있어도 대체로 이상한 징후가 발견되지 않아 바이러스로 인해 피해가 많았다. 본 연구는 사과 왜성대목 M.9 및 M.26의 무독묘 생산을 위하여 고온처리($37^{\circ}C$, 6주), 화학처리(Ribavirin) 및 생장점 배양하여 바이러스 제거 처리를 하였다. 바이러스 검출에 일반적으로 사용되는 방법은 효소면역 측정법(ELlSA)과 중합효소연쇄반응(RT-PCR)을 이용하였는데, RT-PCR은 ELlSA방법보다 10 ~ 30% 더 민감하였다. 사과 왜성대목 바이러스 검정 결과, 바이러스 제거 효율은 생장점 배양이 가장 높았다. 생장점 배양 후 바이러스 무병묘의 획득율은 30 ~ 40%로 높게 나타났다. 생장점 배양에서 사과 왜성대목 M.9은 ACLSV, ASPV 및 ASGV의 비율이 각각 45%, 60%, 50%로 높았고, 사과 왜성대목 M.26에서는 ACLSV, ASPV 및 ASGV의 감염율은 각각 40%, 55%, 55%였다. 이상의 결과, 사과 왜성대목에서 무독묘를 생산할 수 있는 가장 효과적인 방법은 생장점 배양에 의한 것으로 판단되었다.