• Title/Summary/Keyword: viroid

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Detection of Viroid-like RNA Molecules in Korean Peonies (Paeonia lactiflora) (한국산 작약(Paeonia lactiflora)으로부터 바이로이드 유사 RNA 분자의 검출)

  • ;H. L. S nger
    • Korean Journal Plant Pathology
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    • v.13 no.1
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    • pp.1-4
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    • 1997
  • Viroid-like RNA molecules were detected from the low molecular weight RNAs isolated from the Korean peonies which showed typical viroid symptoms of epinasty and dwarfing. Low molecular weight RNAs including viroid RNA molecules were purified by the Qiagen anion exchange minicolumns. Viroid-like RNA molecules showed a single viroid specific band in the native polyacrylamide gel. They were separated into two bands in the denaturing gel conditions. The band of circular form of viroid-like RNAs was crossed over the horizontal band of the linear form of viroid-like RNA molecules in 0~8 M urea gradient gel under the denaturing conditions of 37$^{\circ}C$. The two circular forms of viroid-like RNA molecules were detected in the reverse polyacrylamide gel electrophoresis. The viroid-like RNA molecules purified from the peonies were supposed to be unidentified viroid RNA molecules.

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Viroid-the Smallest Plant Pathogen (바이로이드-가장 작은 식물병원체)

  • Lee Jai Youl
    • Korean Journal Plant Pathology
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    • v.1 no.3
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    • pp.199-206
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    • 1985
  • Viroids are the smallest. well-characterized infectious agents presently known. and so far viroids have been found only in higher plants. The structures of viroid-molecules are single-stranded, covalently closed circular RNA molecules with a range of 240 to 380 nucleotides according to the various viroids. Viroids are remarkable not only as a new category of pathogen, which cause economically important diseases, but also as an excellent model system for biochemical and biophysical investigations because of their small size, relative stability and their self-replication. Four different patato spindle tuber viroid isolates, which express the different symptoms on the same host plant exchange only 2 to 6 nucleotides in the total number of 359 nucleotides, but now the mechanism of viroid pathogenicity is not explained fully. Viroid-melecules are replicated without any special viroid-associated proteins, and during the process of viroid replication oligomeric viroid-associated RNAs are detected at nuclei of viroid infected leaf tissue. The mechanism of viroid replication can now be illustrated according to a possible explanation of rolling-circle system. Although the rapid progress have been made in elucidation of the biochemical and biophysical properties of PSTV and other viroids, the mechanism of viroid replication and pathogenicity is less known and is still a matter of speculation. When these problems can be sufficiently explained, the viroid molecule could play an important role as an available vector in plant genetic engineering.

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Detection and Distribution of Apple scar skin viroid-Korean Strain (ASSVd-K) from Apples Cultivated in Korea

  • Lee, Jai-Youl;Kwon, Mi-Jo;Hwang, Seung-Lark;Lee, Sung-Joon;Lee, Dong-Hyuk
    • The Plant Pathology Journal
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    • v.18 no.6
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    • pp.342-344
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    • 2002
  • Apple scar skin viroid (ASSVd) has been one of the most destructive diseases in Korean apple orchards. Symptoms of the scar skin viroid disease were detected in various apple cultivars, namely, Sansa, Fuji, Chukwang, Miki-Life, Hongro, and Songbongeum cultivated in the southern part of Korea. The RNA molecules were extracted from the apples bearing dapple apple symptoms with the application of CF-11 RNA extraction method. The purified RNAs were used for the synthesis of cDNA with RT-PCR. The PCR products were cloned and sequenced. The viroid RNA molecules from the six different cultivars bearing the dapple symptos showed the same nucleotide sequences as that of the Korean strain of ASSVd(ASSVd-K). ASSVd-K was detected from apple orchards in Kunwi, Sangju, Uiseong, Yeong-yang, Andong, and Youngduk in Gyeongbuk Province in 2001, and in Muju in Jeonbuk Province in 2002. As the viroid disease could be propagated vegetatively, it can be widely transmitted gradually in Korea.

Development of a Quantitative Real-time Nucleic Acid Sequence based Amplification (NASBA) Assay for Early Detection of Apple scar skin viroid

  • Heo, Seong;Kim, Hyun Ran;Lee, Hee Jae
    • The Plant Pathology Journal
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    • v.35 no.2
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    • pp.164-171
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    • 2019
  • An assay for detecting Apple scar skin viroid (ASSVd) was developed based on nucleic acid sequence based amplification (NASBA) in combination with realtime detection during the amplification process using molecular beacon. The ASSVd specific primers for amplification of the viroid RNA and molecular beacon for detecting the viroid were designed based on highly conserved regions of several ASSVd sequences including Korean isolate. The assay had a detection range of $1{\times}10^4$ to $1{\times}10^{12}$ ASSVd RNA $copies/{\mu}l$ with reproducibility and precision. Following the construction of standard curves based on time to positive (TTP) value for the serial dilutions ranging from $1{\times}10^7$ to $1{\times}10^{12}$ copies of the recombinant plasmid, a standard regression line was constructed by plotting the TTP values versus the logarithm of the starting ASSVd RNA copy number of 10-fold dilutions each. Compared to the established RT-PCR methods, our method was more sensitive for detecting ASSVd. The real-time quantitative NASBA method will be fast, sensitive, and reliable for routine diagnosis and selection of viroid-free stock materials. Furthermore, real-time quantitative NASBA may be especially useful for detecting low levels in apple trees with early viroid-infection stage and for monitoring the influence on tree growth.

Chrysanthemum stunt viroid Induces the Accumulation of Small RNAs Associated with RNA Silencing in Infected Chrysanthemum

  • Chung Bong-Nam;Choi Gug-Seoun;Kim Ki-Taek
    • The Plant Pathology Journal
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    • v.22 no.3
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    • pp.235-238
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    • 2006
  • Chrysanthemum stunt viroid (CSVd) induced systemic symptoms on chrysanthemum. We detected small RNAs of approximately 22 nucleotides with sequence specificity to CSVd in chrysanthemum infected with CSVd: an indication of the presence of RNA silencing. Regardless of symptom differences associated with CSVd, the small RNAs distributed similarly in amount. Small RNAs were detected with partial-length or full-length probes, indicating that they are not restricted to specific viroid regions but likely representing most of the viroid molecule.

Incidence of Coleus blumei viroid 1 in Seeds of Commercial Coleus in Korea

  • Chung, Bong-Nam;Choi, Gug-Sun
    • The Plant Pathology Journal
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    • v.24 no.3
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    • pp.305-308
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    • 2008
  • A viroid was detected from symptomless Coleus blumei cultivar 'Kong Scarlet'. It consisted of 249 nucleotides(GenBank accession no. EU410620), which was 100% identical to a Coleus blumei viroid 1(CbVd 1) reported from China(GenBank accession no. DQI78399), indicating that the viroid was an isolate of CbVd1. Attempts were made to determine if commercial Coleus seeds were infected with CbVd. Infection rates in seedlings of the 14 commercial cultivars of Coleus ranged from 0 to 100%. CbVd1 caused discoloration and growth retardation in some cultivars, but is symptomless in others. These results indicated that Coleus in commercial markets in Korea is highly infected with CbVd.

Detection and Genomic Analysis of Viroid-like RNA Molecules Isolated from Korean Peonies (한국산 작약에서 분리한 바이로이드 유사 RNA 분자의 확인과 유전자 분석)

  • 정동수;김무인;이재열
    • Korean Journal Plant Pathology
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    • v.13 no.2
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    • pp.113-117
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    • 1997
  • Low moleuclar weight (LMW) RNAs were isolated form Korean peonies which expressed symptoms of stunt and epinasty. The LMW plant RNAs were purified by Qiagen column chromatography which could separate viroid specific nucleic acid at differential salt concentration. After the inoculation of the purified RNAs from the peonies, the inoculated tomatoes (cv. Rutgers) expressed the symptoms of stunt and epinasty. Also the same molecular weight RNAs with viroid-like RNAs were isolated from the inoculated tomatoes. Double-stranded cDNA were synthesized by the methods of reverse transcription (RT) and polymerase chain reaction (PCR) with the purified RNA and primers. The same cDNAs associated with viroid-like RNAs wre cloned from the inoculated tomatoes. The cDNA has been sequenced and its 375-nucleotides were arranged into secondary structure. The cloned cDNA showed 47~54% homology compared with other viroids. The sequence homology of the cloned cDNA were partially high with plant genomic RNAs.

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Current occurrence of persimmon viroid and citrus viroid in persimmon in JellaNam-do and testing for viroid inactivation methods (전라남도 지역 감 바이로이드의 감염상황 및 무병화 효율 연구)

  • Kim, Dae Hyun;Kim, In-Soo;Lee, Gunsup;Cho, In-Sook;Cho, Kang Hee;Shin, Il Sheob;Kim, Se Hee;Chun, Jae An;Choi, In-Myung
    • Journal of Plant Biotechnology
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    • v.42 no.1
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    • pp.43-48
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    • 2015
  • It is a serious situation that the farmers' income has gradually decreased due to the decline of productivity of fruit trees infected with viroids. It has been known that Persimmon viroid (PVd) and Citrus viroid (CVd) are economically important viroids that can infected persimmon. In this study, the incidence of CVd and PVd in 'Fuyu' persimmon were identified as 41% and 34% in JeollaNam-do, respectively. The collected persimmon samples infected by both PVd and CVd were used for testing efficiency of the viroid inactivation methods. The samples were subjected to single treatment of the heat treatment ($37^{\circ}C$), cold treatment ($4^{\circ}C$), or antiviral agent treatment (Ribavirin), and double treatment of combinations of the three methods. Viroid inactivation efficiency was confirmed through RT-PCR. In the case of the samples subjected to cold treatment for 4 weeks, the viroid inactivation efficiency was most significantly high as 67% against the survival rate of 100%. In addition, in the case of the samples treated for 2 weeks with the antiviral agents and cold treatment, the viroid inactivation rate was similar to that of the cold treatment. In conclusion, the cold treatment showed the highest viroid inactivation efficiency, and this result will provide valuable information for production of viroid-free persimmon.

The Effects of Virus and Viroid Infections on the Growth and Fruit Quality of 'Hongro' Apples (사과 바이러스와 바이로이드 감염이 '홍로' 사과의 수체 및 과실 특성에 미치는 영향)

  • Sang-Yun Cho;Hyun Ran Kim;Kang Hee Cho;Se Hee Kim;Byeonghyeon Yun;Sewon Oh;Ji Hae Jun
    • Korean Journal of Plant Resources
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    • v.37 no.4
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    • pp.401-410
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    • 2024
  • This study was undertaken to elucidate the effects of virus and viroid infections on the growth of trees and the attributes of fruit quality in 'Hongro' apples. Trials were initiated using virus-infected, viroid-infected, combined virus/viroid-infected, and uninfected apple trees in an experimental apple orchard at the National Institute of Horticultural and Herbal Science in 2019. The growth of each tree was measured annually and compared between virus-free and virus/viroid-infected trees. Fruits were harvested from all apple trees, and selected attributes of fruit quality, including yield, weight, firmness, titratable acidity, and anthocyanin content, were determined in September 2021-2022. The results revealed significant differences among virus-free trees and those infected with either virus, viroid, or a combination of virus and viroid. Infection with viral and viroid diseases led to reductions in tree height (14.0%), trunk area (23.1%), fruit yield (65.0%), fruit weight (34.4%), and anthocyanin content (39.8%), while increasing fruit firmness (33.2%) and titratable acidity (39.8%), respectively. We anticipate that our research findings will also be beneficial for apple virus and viroid disease control, as well as apple cultivation management.

Occurrence of Apple scar viroid-Korean strain (ASSVd-K) in Apples Cultivated in Korea

  • Lee, Ju-Hee;Park, Jean-Kyung;Lee, Dong-Hyuk;Uhm, Jae-Youl;Ghim, Sa-Youl;Lee, Jai-Youl
    • The Plant Pathology Journal
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    • v.17 no.5
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    • pp.300-304
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    • 2001
  • Apple is the most economically important fruit in Korea. The suspected viroid disease of dapple apple was found in apple fruits cultivated in Kyungpook province. Symptoms begin in mid-July as small circular spots, which stand out against the background color on the young fruit. Dappling of the fruit becomes more intense and easier to detect as the fruit approaches maturity; the affected spots remain yellowish as the fruit matures. no leaf or bark syndromes have been associated with this disease. The infected fruits are downgraded considerably during quality grading. The low molecular weight RNA containing viroid RNA molecules were extracted from the peels of the apples with dapple symptoms. The RNA molecules were extracted from the apples using Qiagen column chromatography. The purified RNAs were used for the synthesis of cDNA with RT-PCR. The PCR products were then ligated into a pGEM-T Easy vector, cloned and sequenced. The sequence of the viroid RNA molecule shows 331 nucleotides with one base difference ("G" insertion between the position of 133 and 134) compared with that of the Apple scar skin viroid (ASSVd) reported by Hashimoto and Koganezawa in Japan. This is the first report on the occurrence of the ASSVd in apple trees cultivated in Korea, as well as the identification of a new Korean strain of the ASSVd.the ASSVd.

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