• Proceedings of the Korean Society of Plant Pathology Conference
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• 1999.10a
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• pp.18.1-18
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• 1999

• Journal of Life Science
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• v.26 no.5
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• pp.620-631
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• 2016

Viroids are about 250-400 base pair of short single strand RNA fragments have been associated with economically important plant diseases. Due to the lack of protein expression capacity associated with replication, it is very difficult to diagnosis viroid diseases in serological methods. For detecting viroid at plants, molecular-based techniques such as agarose gel electrophoresis, polyacrylamide gel electrophoresis (PAGE), DNA-hybridization, blotting analysis and conventional RT-PCR are reliable. Real-time RT-PCR methods that grafted on RT-PCR methods with improved confirmation methods have been also utilized. However, they are still labor-intensive, time-consuming, and require personnel with expertise. Loop-mediated Isothermal Amplification (LAMP) method is a nucleic acid amplification method under the isothermal condition. The LAMP methodology has been reported to be simple, rapid, sensitive and field applicable in detecting a variety of pathogens. The results of LAMP method can be colorized by adding a visible material such as SYBR green I, Evagreen, Calcein, Berberine and Hydroxy naphthol blue (HNB) with simple equipment or naked eyes. The combination of LAMP method and nucleic pathogens, viroids, can be used to realize simple diagnosis platform for the genetic point-of care testing system. The aim at this review is to summary viroid-caused diseases and the simple visible approach for diagnosing viroids using Loop-mediated Isothermal Amplification (LAMP) method.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2002.11a
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• pp.125.1-125
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• 2002

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1999.10a
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• pp.71.1-71
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• 1999

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.143.1-143
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• 2003

Hop stunt viroid(HSVd) is a plant pathogen which infect a number of hosts such as grapevine, Citrus and Prunus plants. Sequence variants of HSVd have been divided into three types(i. grapevine and hop, ii. citrus, iii. plum, peach, apricot and almond). Purified RNAs from plum trees were used for the synthesis of cDNA with reverse transcription and amplified by polymerase chain reaction. Cloned cDNAs were sequenced and two different consensus sequence variants were detected. A neighbor-joining analysis was carried out on the sequence variants together with 62 previously described variants of HSVd from hop, plum and other species. Sequence variants from plum trees cultivated in Korea were clustered in HSVd-plum subtype and not in HSVd-hop subtype which were two Korean isolates belongs. These relationship between sequence variants from plum and two Korean isolates in HSVd-hop type supports the other origin for hop stunt disease.

• The Plant Pathology Journal
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• v.22 no.4
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• pp.375-380
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• 2006

In the survey from 1992 to 2000, twenty-eight parasitic diseases were observed in major apple producing areas in Korea. The predominant apple diseases were white rot(Botryosphaeria dothidea), Marssonina blotch(Marssonina mali), Valsa canker(Valsa ceratosperma), Alternaria leaf spot(Alternaria mali), and bitter rot(Collectotrichum gloeosporioides and C. acutatum). Apple scab that reappeared in 1990 after disappearance for 15 years was disappeared again since 1997. A viroid disease(caused by apple scar skin viroid) was newly found in this survey. The five diseases, fire blight(Erwinia amylovora), black rot(Botryosphaeria obtusa), scab(Cladosporium carpophilum), Monochaetia twig blight(Monochaetia sp.), and brown leaf spot(Hendersonia mali), which had once described in 1928 but no further reports on their occurrence, were not found in this survey. However, blossom blight(Monilinia mali), brown rot(Monilinia fructigena), and pink rot(Trichothecium roseum), which did not occur on apple after mid 1970s, were found in this survey.

• Korean Journal of Plant Tissue Culture
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• v.27 no.5
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• pp.379-385
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• 2000

tRNA and 7SL RNA based antisense vehicles were prepared by inserting conserved anti-viral and anti-viroid domains. Anti-PVS coat protein leader sequence (ACPL) and antistructural antihairpin domain of PSTVd (AHII) were inserted in tRNA cassette; anti- zing finger domain of PVS, AHII and anti hop latent viroid ribozyme were inserted in 7SL RNA gene isolated from A. thaliana. These constructs were shown to be transcribed both, in in vitro and in in vivo conditions. However, it followed from our work that closely linked position of PoIII reference genes and PoIIII antisense genes within T-DNA lead to the impairment of RNA expression in transgenic plants. To assay in vivo transcription of antisense genes, hairy root potato cultures were established using h. tumefaciens A4-24 bearing both, Ri plasmid and PoIII-promoterless plant expression vectors with antisense RNA genes. Expression of antisense RNA in transgenic potato tissues was proven by specific RT-PCR reactions.

• The Plant Pathology Journal
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• v.22 no.2
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• pp.152-154
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• 2006

CSVd-infected chrysanthemum plants grown under $10^{\circ}C\;or\;15^{\circ}C$ growth chamber for 2 months resulted a higher dilution endpoint of template RNA for Reverse transcription and polymerase chain reaction (RT-PCR) than those grown for 1 month: $10^{-4}(1.35{\times}10^{-2}ug/ml)$ for 1 month, and $10^{-3}(1.35{\times}10^{-1}ug/ml)$ for 2 months. Independent experiment, shoots cut from CSVd (Chrysanthemum stunt viroid)-in footed chrysanthemum plants grown under $10^{\circ}C\;or\;20^{\circ}C$ growth chamber for 2 months showed the same CSVd concentration as control($30^{\circ}C$) at 8 weeks after moving them to normal green-house condition($30^{\circ}C$). From those results, it was concluded that even though the concentration of CSVd was reduced in plants grown at low temperatures, when they were moved to normal glass-house temperature CSVd concentration increased to that of untreated plants in 8 weeks. This conclusion was supported by the rapid replication of CSVd in chrysanthemum after infection.

• Research in Plant Disease
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• v.19 no.4
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• pp.288-293
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• 2013

In worldwide, viral diseases of apple plants has caused the serious problems like reduced production and malformation of fruits. Also, the damages of apple plants by virus and/or viroid infection (Apple chlorotic leaf spot virus, Apple stem grooving virus, Apple mosaic virus, and Apple scar skin viroid) were reported in Korea. However there is few report about the protection approach against the infection by apple viruses. Therefore, this paper introduced the experimental protocol for the development of virus-free apple cultivars (Danhong, Hongan, Saenara, Summerdream). Apple plants were treated at $37^{\circ}C$ for 4 weeks and shoot tips were cultured in vitro. After heat treatment, the detection of apple viruses was performed by RT-PCR using virusspecific detection primers in new apple cultivars. With the heat treatments followed by in vitro shoot tip culture, the proportion of virus-free stocks of 'Danhong', 'Hongan', 'Saenara', and 'Summerdream' was 28%, 16%, 12%, and 12%, respectively. Taken together, this approach can be a good tool for production of virus-free apple stocks.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.54-54
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• 2019

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. In many cases, apple is infected with virus and viroid with no specific symptoms, the damage caused by the virus are unaware significantly. In our research, we tried to eliminate viruses in the rootstock for the disease-free seedlings of the apple dwarfing rootstock M.9 and M.26. The method of virus elimination was meristem culture, heat($37^{\circ}C$, 6weeks) treatment and chemistry($Ribavirin^{(R)}$) treatment. The analytical methods commonly used for the detection of virus is Enzyme-linked Immuno-Sorbent Assay(ELlSA) and Reverse Transcription-polymerase Chain Reaction(RT-PCR). RT-PCR method was more 30% sensitive than ELISA method. Efficiency of method eliminate virus appeared meristem method > heat treatment > chemistry treatment. The higher acquisition rate of disease-free seedlings is 30~40% on meristem treatment. In meristem treatment, the apple dwarfing rootstock M.9 gained infection ratio of ACLSV, ASPV and ASGV were 45%, 60% and 50% respectively. In the apple dwarfing rootstock M.26, infection ratio of ACLSV, ASPV and ASGV were 40%, 55%, 55%, respectively. Based on our results, it was found that most effective method of disease-free seedlings apple dwarfing rootstocks was by meristem treatment than heat method and chemistry treatment.