• The Plant Pathology Journal
• /
• 제18권5호
• /
• pp.259-265
• /
• 2002

Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was detected and isolated from diseased 'Fuji' apple (Malus domestica) in Korea. The coat protein (CP) genes of two ApMV strains, denoted as ApMV-Kl and ApMV-K2, were amplified by using the reverse transcription and polymerase chain reaction (RT-PCR) and were analyzed thereafter. The objectives were to define the molecular variability of genomic information of ApMV found in Korea and to develop virus-derived resistant gene source for making virus-resistant trans-genic apple. RT-PCR amplicons for the APMVS were cloned and their nucleotide sequences were determined. The CPs of ApMV-Kl and ApMV-K2 consisted of 222 and 232 amino acid residues, respectively. The identities of the CPs of the two Korean APMVS were 93.1% and 85.6% at the nucleotide and amino acid sequences, respectively. The CP of ApMV-Kl showed 46.1-100% and 43.2-100% identities to eight different ApMV strains at the nucleotide and amino acid levels, respectively. When ApMV-PV32 strain was not included in the analysis, ApMV strains shared over 83.0% and 78.6% homologies at the nucleotide and amino acid levels, respectively. ApMV strains showed heterogeneity in CP size and sequence variability. Most of the amino acid residue differences were located at the N-termini of the strains of ApMV, whereas, the middle regions and C-termini were remarkably conserved. The APMVS were 17.(1-54.5% identical with three other species of the genus Ilarviyus. ApMV strains can be classified into three subgroups (subgroups I, II, and III) based on the phylogenetic analysis of CP gene in both nucleotide and amino acid levels. Interestingly, all the strains of subgroup I were isolated from apple plants, while the strains of subgroups II and III were originated from peach, hop, or pear, The results suggest that ApMV strains co-evolved with their host plants, which may have resulted in the CP heterogeneity.

• Journal of Plant Biotechnology
• /
• 제49권1호
• /
• pp.46-60
• /
• 2022

The genetic variability and population structure of apple mosaic virus (ApMV) have been studied; however, synonymous codon usage patterns influencing the survival rates and fitness of ApMV have not been reported. Based on phylogenetic analyses of 52 ApMV coat protein (CP) sequences obtained from apple, pear, and hazelnut, ApMV isolates were clustered into two groups. High molecular diversity in GII may indicate their recent expansion. A constant and conserved genomic composition of the CP sequences was inferred from the low codon usage bias. Nucleotide composition and relative synonymous codon usage (RSCU) analysis indicated that the ApMV CP gene is AU-rich, but G- and U-ending codons are favored while coding amino acids. This unequal use of nucleotides together with parity rule 2 and the effective number of codon (ENC) plots indicate that mutation pressure together with natural selection drives codon usage patterns in the CP gene. However, in this combination, selection pressure plays a more crucial role. Based on principal component analysis plots, ApMV seems to have originated from apple trees in Europe. However, according to the relative codon deoptimization index and codon adaptation index (CAI) analyses, ApMV exhibited the greatest fitness to hazelnut. As inferred from the results of the similarity index analysis, hazelnut has a major role in shaping ApMV RSCU patterns, which is consistent with the CAI analysis results. This study contributes to the understanding of plant virus evolution, reveals novel information about ApMV evolutionary fitness, and helps find better ApMV management strategies.

• 식물병연구
• /
• 제23권4호
• /
• pp.355-362
• /
• 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.

• 식물병연구
• /
• 제26권2호
• /
• pp.95-102
• /
• 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.

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

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

• The Plant Pathology Journal
• /
• 제19권3호
• /
• pp.159-161
• /
• 2003

The coat protein (CP) gene of Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was selected for the design of virus-specific primers for amplification and molecular detection of the virus in cultivated apple. A combined assay of reverse transcription and polymerase chain reaction (RT-PCR) was performed with a single pair of ApMV-specific primers and crude nucleic acid extracts from virus-infected apple for rapid detection of the virus. The PCR product was verified by restriction mapping analysis and by sequence determination. The lowest concentration of template viral RNA required for detection was 100 fg. This indicates that the RT-PCR for detection of the virus is a 10$^3$times more sensitive, reproducible and time-saving method than the enzyme-linked immunosorbent assay. The specificity of the primers was verified using other unrelated viral RNAs. No PCR product was observed when Cucumber mosaic virus (Cucumovirus) or a crude extract of healthy apple was used as a template in RT-PCR with the same primers. The PCR product (669 bp) of the CP gene of the virus was cloned into the plasmid vector and result-ant recombinant (pAPCP1) was selected for molecule of apple transformation to breed virus-resistant transgenic apple plants as the next step. This method can be useful for early stage screening of in vitro plantlet and genetic resources of resistant cultivar of apple plants.

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

• 한국식물병리학회:학술대회논문집
• /
• 한국식물병리학회 1999년도 임시총회 및 추계 학술발표회
• /
• pp.69.1-69
• /
• 1999

• 식물병연구
• /
• 제18권4호
• /
• pp.391-395
• /
• 2012

국내에서 재배되는 복숭아에 발생하는 바이러스병을 조사하기 위하여 경북 영천 등 복숭아 주산단지 6개 지역에서 전체 시료 515점을 채집하여 5종(ACLSV, ApMV, PDV, PNRSV, PPV) 바이러스의 감염여부를 RT-PCR 방법을 이용하여 바이러스 검정을 실시하였다. RT-PCR 검정 결과 전체 시료 515점 중 335점의 시료에서 ACLSV와 PNRSV가 검출되어 검정한 시료의 65.0%가 바이러스에 감염된 것을 확인하였다. ACLSV가 검출된 복숭아나무는 잎에 모자이크 증상이 관찰되었으며 PNRSV가 검출된 복숭아나무는 별다른 이상증상이 관찰되지 않았다. 검출된 바이러스의 유전자 염기서열 분석으로 ACLSV 4개와 PNRSV 3개의 분리주들을 확인하였으며 ACLSV 분리주들 간에는 95% 이상, PNRSV는 88% 이상의 아미노산 서열 상동성을 나타냈다. 기존에 보고된 ACLSV 및 PNRSV 분리주들과의 아미노산 서열 비교에서는 ACLSV 분리주들의 경우 70-99%, PNRSV 분리주들의 경우 88-99%의 상동성을 보였다. 이들 바이러스 분리주들의 계통학적 분석에서 ACLSV 분리주들은 A, B 그룹 중 A그룹에, PNRSV 분리주들은 I (PV32), II (PV96), III (PE5) 그룹에 각각 하나씩 속하는 것으로 나타났다.