• 식물병연구
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• 제23권3호
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• pp.288-293
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• 2017

Chrysanthemum chlorotic mottle viroid (CChMVd) fused to the leader sequence of a reporter gene (mRFP) expressed transiently in agroinfiltrated Nicotiana benthamiana, was used to show that CChMVd can traffic into chloroplasts, thought to be the site of its replication. Fluorescence from mRFP was detected in chloroplasts, but only if the viroid transcription fusions were present, either from the full-length 400-nt CChMVd, or each of two partial fragments (nucleotides 125 to 2 and 231 to 372). The mRFP and its mRNA were detected by western blotting and RT-PCR, respectively, in tissue extracts of plants infiltrated by each fusion construct. Isolated chloroplasts were shown by RT-PCR to contain the RNA sequences of both CChMVd and mRFP, if both were present, but not the mRFP sequence in the absence of the viroid sequences. The results suggest that RNA trafficking was probably due to an RNA structure, and not a particular sequence, as discussed.

• 한국식물병리학회지
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• 제14권6호
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• pp.612-613
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• 1998

The nucleotide sequence of hop stunt viroid HHSVd) Kh strain was sequenced by the reverse transcription and polymerase chain reaction. It consists of 296 nucleotides, and differs by one nucleotide deletion of cytosine at the position of 295 from the HSVd-K strain which consists of 297 nucletoides.

• The Plant Pathology Journal
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• 제22권4호
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• pp.334-338
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• 2006

Chrysanthemum chlorotic mottle viroid(CChMVd) isolates have been identified from chrysanthemum showing yellow spots or infected without symptom. They were 399-400 nucleotides length of RNA. CChMVd-SSHA6(GenBank accession no. DQ450682) revealed a GAAA to DUUC substitution in positions 82-85 of CChMVd-MSIN34(GenBank accession no. DQ402041). In vitro RNA transcripts with the complete CChMVd sequence were infectious and induced the typical CChMVd infection symptom of yellow spots in chrysanthemum cv. Sharotte. CChMVd caused reduction in growth in some cultivars, whereas some cultivars were not affected. This is the first report on the occurrence of CChMVd in chrysanthemum 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.

• The Plant Pathology Journal
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• 제30권1호
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• pp.68-74
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• 2014

Chrysanthemum stunt viroid (CSVd), a noncoding infectious RNA molecule, causes seriously economic losses of chrysanthemum for 3 or 4 years after its first infection. Monomeric cDNA clones of CSVd isolate SK1 (CSVd-SK1) were constructed in the plasmids pGEM-T easy vector and pUC19 vector. Linear positive-sense transcripts synthesized in vitro from the full-length monomeric cDNA clones of CSVd-SK1 could infect systemically tomato seedlings and chrysanthemum plants, suggesting that the linear CSVd RNA transcribed from the cDNA clones could be replicated as efficiently as circular CSVd in host species. However, direct inoculation of plasmid cDNA clones containing full-length monomeric cDNA of CSVd-SK1 failed to infect tomato and chrysanthemum and linear negative-sense transcripts from the plasmid DNAs were not infectious in the two plant species. The cDNA sequences of progeny viroid in systemically infected tomato and chrysanthemum showed a few substitutions at a specific nucleotide position, but there were no deletions and insertions in the sequences of the CSVd progeny from tomato and chrysanthemum plants.

• 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.

• The Plant Pathology Journal
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• 제22권1호
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• pp.63-67
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• 2006

Apple scar skin, one of the most destructive diseases affecting apple, is caused by Apple scar skin viroid (ASSV d). Fruit dappling appeared on several cultivars in Korea and has been distributed to major cultivated areas since 2001. ASSVd was identified from infected fruits by using nucleic acid sequence-based amplification with electrochemiluminescence (NASBA-ECL). NASBA-ECL method was faster and hundredfold more sensitive than reverse transcription-polymerase chain reaction (RT-PCR) for ASSVd detection in apple leaves/ stems. ASSVd was rapidly transmitted to the entire tree in the second year after artificial inoculation. The ASSVd could be transmitted efficiently by using contaminated pruning scissors to both lignified stems (60 to $70\%$) and green shoots (20 to $40\%$) of apple tree and young plants. Dipping of contaminated scissors in $2\%$ sodium hypochlorite solution effectively prevented viroid transmission. In the ASSV d-infected fruits, the viroid was easily detected from fruit skin, seed coat, and embryo. Moreover, embryo and endosperm separately excised from the ASSVd-infected seeds were ASSVd positive in NASBA-ECL assay. Seedlings germinated from ASSVd-positive seeds showed $7.7\%$ infection rate., which indicated that ASSVd is seed-borne.

• 식물병연구
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• 제21권4호
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• pp.346-350
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• 2015

국내에서 재배되는 사과 '홍로'에 발생하는 Apple scar skin viroid병의 유전자 계통분석을 하기 위하여 바이로이드 증상이 나타나는 8개 지역(봉화, 청송, 당진, 김천, 무주, 문경, 수원, 영월)의 농가에서 수집한 시료를 RT-PCR 방법을 이용하여 바이로이드 검정을 실시하였다. 검출된 바이로이드의 클로닝과 유전자 염기서열 분석을 통해 8종의 분리주들을 확인하였고 한국, 인도, 중국, 일본 및 그리스에서 보고된 21종의 분리주와 염기서열을 비교하였다. 8개의 분리주들의 핵산 서열은 기존에 보고된 분리주들과 92.2-99.7%의 상동성을 나타냈다. 계통분석을 통해 본 연구에서 보고한 7종의 분리주들은 기존의 것들과 독립된 그룹에 속하는 것으로 나타났다.

• The Plant Pathology Journal
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• 제17권4호
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• pp.194-200
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• 2001

Chrysanthemum stunt viroid (CSVd) ws identified in chrysanthemum cv. Chunkwang showing symptoms of stunt with leaf distortion (K1) and stunt with chlorosis of leaves (K2) collected from the main cultivation area of Masan, Kyongnam province in Korea. The specific RNAs related with the diseased chrysanthemums were detected. Full-length 354 bp CSVd cDNAs were amplified from infected tissue by reverse transcription and polymerase chain reaction using a pair of primers specific for CSVd sequence. The amplified cDNA products were analyzed by agarose gel electrophoresis and the specific cDNAs were cloned. Nucleotide sequences of the two CSVd isolates K1 and K2 varied. Phylogenetic analysis of the nucleotide sequences of CSVd isolates indicated that K1 was closely related with J2 and Am 2 isolates. K1 and K2 were transmitted by grafting to Dendranthema grandiflorum cv. Mistletoe, Gynura aurantiaca, and Lycopersicon esculentum cv. Rutgers. This is the first report of CSVd in D. grandiflorum in Korea.

• The Plant Pathology Journal
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• 제24권1호
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• pp.31-35
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• 2008

The presence of Chrysanthemum stunt viroid (CSVd) in seed and pollen of diseased chrysanthemum was demonstrated. In seeds infected male parent from crosses in May, CSVd was transmitted to 6.7% of the progeny seedlings, whereas if the female parent was infected, CSVd transmission rate was between 46.9 and 75.7%. A relatively high incidence of 94.4 to 96.0% seed transmission occurred when both parents were infected. In seeds infected male parent from crosses in December, no progeny seedlings were infected with CSVd, whereas if the female parent did, CSVd transmission rate was 1.5%. When both parents were infected, 6.9% seed transmission was occurred. The seed transmission rate depended on the temperature when the crosses were made. CSVd was not detected in the non-infected female parent pollinated with infected pollen but was transmitted to the progenies. This is the first report of seed-borne transmission of CSVd in chrysanthemum.