• 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.

• The Plant Pathology Journal
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• 제30권2호
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• pp.195-199
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• 2014

Apple stem pitting virus (ASPV), of the Foveavirus genus in the family Betaflexiviridae, is one of the most common viruses of apple and pear trees. To examine variability of the coat protein (CP) gene from ASPV, eight isolates originating from 251 apple trees, which were collected from 22 apple orchards located in intensive apple growing areas of the North Gyeongsang and North Jeolla Provinces in Korea, were sequenced and compared. The nucleotide sequence identity of the CP gene of eight ASPV isolates ranged from 77.0 to 97.0%, while the amino acid sequence identity ranged from 87.7 to 98.5%. The N-terminal region of the viral CP gene was highly variable, whereas the C-terminal region was conserved. Genetic algorithm recombination detection (GARD) and single breakpoint recombination (SBP) analyses identified base substitutions between eight ASPV isolates at positions 54 and 57 and position 771, respectively. GABranch analysis was used to determine whether the eight isolates evolved due to positive selection. All values in the GABranch analysis showed a ratio of substitution rates at non-synonymous and synonymous sites (dNS/dS) below 1, suggestive of strong negative selection forces during ASPV CP history. Although negative selection dominated CP evolution in the eight ASPV isolates, SLAC and FEL tests identified four possible positive selection sites at codons 10, 22, 102, and 158. This is the first study of the ASPV genome in Korea.

• 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.

• Journal of Plant Biotechnology
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• 제44권4호
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• pp.379-387
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• 2017

Apple scar skin viroid (ASSVd), Apple chlorotic leaf spot virus(ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV)는 사과를 감염시키는 대표적인 바이러스로, 이러한 4종 바이러스에 복합 감염된 사과 '홍로'의 동절기 가지의 휴면아(측아)로부터 다양한 크기 (0.4 mm ~ 1.2 mm)와 발달단계 (이하 Stage 1, Stage 2, Stage 3라 칭함)의 경정 및 측부 분열조직을 절취하여 BA 3.0 mg/L 와 IBA 0.1 mg/L가 혼합된 배지에 배양하여 캘러스를 형성시킨 후, 감염 바이러스바이러스의 제거유무를 조사하였다. 직경 10 mm이상 증식된 캘러스 31 계통을 RT-PCR 분석으로 제거효율을 알아 본 결과, ACLSV는 100%, ASSVd와 ASPV는 93.5%의 높은 제거 효율을 보인 반면, ASGV는 25.8%로 상대적으로 제거효율이 낮았다. 4종 바이러스가 동시에 모두 제거되는 경우는 Stage 1의 휴면아의 분열조직을 배양하였을 때만 가능하였는데, 그 이유는 ASGV가 이 시기의 배양에서 주로 제거되었기 때문으로, ASGV는 다른 바이러스와는 달리 분열조직의 발달단계가 큰 영향을 미치는 것으로 나타났다. 휴면아의 분열조직을 배양하여 바이러스를 제거한 예는 세계적으로 보고된 바가 없는 것으로서, 본 연구결과는 향 후 사과의 무병묘 생산에 매우 유용한 자료로 활용될 것이다.

• The Plant Pathology Journal
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• 제20권2호
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• pp.147-154
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• 2004

A simple and reliable procedure for RT-PCR detection of Apple stem pitting virus (ASPV), Cherry rasp leaf virus (CRLV), and Cherry necrotic rusty mottle virus (CNRMV) was developed. Two virus specific primer sets for each virus were found to specifically detect each virus among fourteen sets of designed oligonucleotide primers. Total RNAs extracted from healthy and from ASPV-,CRLV- and CNRMV-infected plant tissues were used to synthesize cDNA using oligo dT primer and then amplified by virus-specific primers for each virus. Each primer specifically amplified DNA fragments of 578 bp and 306 bp products for ASPV (prAS CP-C and prAS CP-N primers, respectively); 697 bp and 429 bp products for CRLV (prCR4 and prCR5-JQ3D3 primers, respectively); and 370 bp and 257 bp products for CNRMV (prCN4 and prCN6-NEG 1 primers, respec-tively) by RT-PCR. DNA sequencing of amplified DNA fragments confirmed the nature of each amplified DNA. Altogether, these results suggest that these virus specific primer sets can specifically amplify viral sequences in infected tissues and thus indicate that they can be used for specific detection of each virus.

• 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.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.52-52
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• 2018

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, furthermore, its damages and economic losses have increased constantly. In our research, we tried to survey virus infection for commercial nursery trees of major apple cultivars, especially dwarfing rootstocks 'M.9' and 'M.26' as well as scions. Trees were collected from 11 locations which have produced a great amount of apple nursery stocks in Korea. Infection degree was investigated in apple cultivars, 'Hongro' and 'Fuji' using RT-PCR method. In the scion of cultivar 'Hongro', infection ratio of ACLSV, ASPV and ASGV were 100%, 81.8% and 100% respectively. In the rootstock of cultivar 'Hongro', infection ratio of ACLSV, ASPV, ASGV and ApMV were 90.9%, 81.8%, 100% and 9.1% respectively. In the scion of cultivar 'Fuji', infection ratio of ACLSV, ASPV and ASGV were 81.8%, 90.9% and 100% respectively. In the rootstock of cultivar 'Fuji', infection ratio of ACLSV, ASPV, ASGV and ApMV were 81.8%, 90.9%, 100% and 9.1% respectively. Infection of ASSVd was not detected in both cultivars. From our results, it was found that most of apple rootstocks and scions had multiple infections by apple viruses which have caused economic damage in fruit production.

• 식물병연구
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• 제27권2호
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• pp.79-83
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• 2021

The aim of the present study was to develop a sensitive and specific detection method for the rapid detection of apple scar skin viroid (ASSVd) in apple leaves. The resulting reverse transcription recombinase polymerase amplification (RT-RPA) assay can be completed in 10 min at 42℃, is 10 times more sensitive than conventional reverse transcription polymerase chain reaction, and can specifically amplify ASSVd without any cross-reactivity with other common apple viruses, including apple stem grooving virus, apple stem pitting virus, and apple chlorotic leaf spot virus. The reliability of the RT-RPA assay was assessed, and the findings suggested that it can be successfully utilized to detect ASSVd in field-collected samples. The RT-RPA assay developed in the present study provides a potentially valuable means for improving the detection of ASSVd in viroid-free certification programs, especially in resource-limited conditions.

• 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.

• 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.