• Horticultural Science & Technology
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• v.32 no.4
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• pp.544-549
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• 2014

Lily symptomless virus (LSV), Lily mottle virus (LMoV), and Cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf and bulb samples showing characteristic symptoms of virus infection were collected in 2012, and 80 field samples were analyzed by reverse transcription polymerase chain reaction (RT-PCR). The infection frequencies were 79% for LMoV, 5% for LSV, and 3% for CMV. The LMoV coat protein gene was amplified and cloned into the pET21d(+) expression vector to develop serological diagnostic tools to detect LMoV. The resulting carboxy-terminal His-tagged coat proteins were expressed in Escherichia coli strain BL21 (DE3) by induction with IPTG. The recombinant proteins were purified using Ni-NTA agarose beads and used as an antigen to produce polyclonal antibodies in laying hens. The resulting egg yolk immunoglobulin (IgY) specifically recognized LMoV from infected plant tissues in immunoblotting assays and had comparable sensitivity to that of a mammalian antibody. In addition, method of immunocapture RT-PCR using this IgY was developed for sensitive, efficient, and rapid detection of LMoV. Based on these results, large-scale bulb tests and detection of LMoV in epidemiological studies can be performed routinely using this IgY. This is the first report of production of a polyclonal IgY against a plant virus and its use for diagnosis.

• Research in Plant Disease
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• v.9 no.4
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• pp.237-241
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• 2003

Damages caused by infection with Cucumber mosaic virus (CMV), Lily mottle virus (LMoV) and Lily symptomless virus (LSV) were assessed by comparing growth of plants produced from seeds of Lilium x fomolongi cultivar 'Noesan' both infected and free from infection with those viruses. Symptoms and infection rate were investigated in field-grown lily.Dominant viral infection symptom in the field was mottle on leaves, caused by natural infection with LMoV. Incidence of viral disease caused by mixed infection with two of LMoV, CMV and LSV in field-grown Lilium x fomolongi cultivars, reused for more than 6 years consecutively, was 80 percent. In comparison with healthy Lilium x fomolongi cultivar 'Noesan', plants doubly infected with CMV-Li1 and LMoV-Li diminished their plant height by 14 percent, fresh weight by 38 percent, and flower length by 15 percent. Lily plants singly infected with CMV-Li1 or LMoV-Li significantly reduced their freish weight by 21.8% and 28.4% compared to healthy plants, respectively.

• Korean Journal of Microbiology
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• v.45 no.3
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• pp.251-256
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• 2009

Leaf samples and bulbs showing characteristic symptoms of virus infection were collected from Gang-won, Chung-nam, and Jeju Province of Korea in 2008-2009. Three viruses, Lily symptomless virus (LSV), Lily mottle virus (LMoV), and Cucumber mosaic virus (CMV) were detected by RT-PCR. Virus-infected plant samples were identified; 12 plants with LSV, 20 plants with LMoV, and 1 plant with CMV. Of the twelve LSV infected samples, seven samples were found to be mix-infected with LMoV and LSV. Symptoms of LMoV and LSV mixed infection were fairly severe, like as vein clearing, leaf curling, leaf mottling, leaf mosaic, and yellow streaking. Mixed infection with LMoV and LSV was also found in lily bulbs which have been stored under unfavorable environmental conditions. LMoV predominated in our tests, whereas spread of Lilyvirus X (LVX) was not found. The nucleotide sequences of coat protein (CP) region of seven isolates (4 LMoV, 2 LSV, and 1 CMV) were compared with the corresponding regions of LMoV (AJ564636), LSV (AJ516059) and CMV(AJ296154). The nucleotide sequence homologies between reference viruses and seven isolates were 95-99%. Complete sequencing of seven isolates is necessary to obtain more information on the molecular characteristics of these viruses as well as to increase sensitivity and rapidity of viral detection.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.107.2-108
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• 2003

Viral disease symptoms were investigated in the field grown Longiflorum hybrid cultivars, and the damages caused by infection with Lily mottle virus (LMoV) and Cucumber mosaic virus (CMV) were assessed by comparing growth of plants produced from seeds of Longiflorum hybrid cultivar both infected by artificial inoculation and free from infection with theses viruses. Dominant symptom caused by spotaneous infection with LMoV and CMV in the field was mottle combined with chlorotic stripe on leaves. LMoV developed brownish necrotic lesion on floral leaves. The incidence of viral disease by mixed infection with LMoV, CMV or Lily symptomless virus (LSV) in the filed grown Longiflorum hybrid cultivar, cultivated for more than 6 years, was 80 to 84 percent. In comparison with virus-free plants, plants doubly infected with CMV and LMoV by artificial inoculation decreased stem length by 14 percent and fresh weight by 38 percent. In conclusion, flower quality and the stem length of Longiflorum hybrid cultivar were affected by LMoV and CMV infection.

• The Plant Pathology Journal
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• v.36 no.2
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• pp.170-178
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• 2020

The Lily mottle virus (LMoV) impedes the growth and quality of lily crops in Lanzhou, China. Recently Arabis mosaic virus (ArMV) has been detected in LMoV-infected plants in this region, causing plant stunting as well as severe foliar symptoms, and likely posing a threat to lily production. Consequently, there is a need to develop simple, sensitive, and reliable detection methods for these two viruses to prevent them from spreading. Reverse transcription (RT) loop-mediated isothermal amplification (LAMP) assays have been developed to detect LMoV and ArMV using two primer pairs that match six conserved sequences of LMoV and ArMV coat proteins, respectively. RT-LAMP assay results were visually assessed in reaction tubes using green fluorescence and gel electrophoresis. Our assays successfully detected both LMoV and ArMV in lily plants without the occurrence of viral cross-reactivity from other lily viruses. Optimal conditions for LAMP reactions were 65℃ and 60℃ for 60 min for LMoV and ArMV, respectively. Detection sensitivity for both RT-LAMP assays was a hundredfold greater than that of our comparative RT-polymerase chain reaction assays. We have also found this relatively rapid, target specific and sensitive method can also be used for samples collected in the field and may be especially useful in regions with limited or no laboratory facilities.

• Research in Plant Disease
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• v.25 no.2
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• pp.79-83
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• 2019

To investigate the incidence status of lily viruses on Jeju island, lily samples were collected from 2015 to 2018 and examined for virus infection using RT-PCR. Of the viral infections, mixed and single infections were 70.0% and 17.9%, respectively. The incidence of mixed infections was highest for PlAMV and LSV as 43.4% in 2015; PlAMV, LSV 33.1% in 2016; LSV, LMoV 10.2% in 2017; and PlAMV, LSV, LMoV and CMV 15.8% in 2018. The incidence of PlAMV was observed to be 82.0% in 2015, 49.4% in 2016, 13.6% in 2017, and 39.5% in 2018 after the first occurrence of PlAMV in 2013. No symptoms were observed for single infection with LSV. However, in the case of mixed infection with LSV and LMoV, mosaic and leaf malformation symptoms appeared. With mixed infection with LSV and CMV, pale brown necrotic spots appeared, and mosaic and leaf curling were induced. PlAMV was more common in mixed infection than in single infection, and caused necrosis following the development of reddish-brown spots. PlAMV significantly decreased the marketability of lilies owing to the generation of leaf anomalies and curls, and its symptoms were more severe in mixed infections.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1776-1781
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• 2012

Multiple viruses such as Lily symptomless virus (LSV), Lily mottle virus (LMoV), and cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf samples and bulbs showing characteristic symptoms of virus infection were collected from Gangwon, Chungnam, and Jeju provinces of Korea in 2008-2011. Coat protein (CP) genes of LSV and LMoV were amplified from collected samples by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into a pET21d(+) expression vector to generate recombinant CPs. The resulting carboxy-terminal His-tagged CPs were expressed in Escherichia coli strain BL21(DE3) by isopropyl-1-thio-${\beta}$-D-galactoside induction. The recombinant proteins were purified using Ni-NTA agarose beads, and the purified proteins were used as an immunogen to produce polyclonal antibodies in rabbits. The resulting polyclonal antisera recognized specifically LSV and LMoV from infected plant tissues in Western blotting assays. Indirect enzymelinked immunosorbent assay and immunocapture RT-PCR using these polyclonal antisera were developed for the sensitive, efficient, economic, and rapid detection of Lily viruses. These results suggest that large-scale bulb tests and economic detection of Lily viruses in epidemiological studies can be performed routinely using these polyclonal antisera.

• The Plant Pathology Journal
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• v.25 no.3
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• pp.225-230
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• 2009

Lily mottle virus (LMoV) causes flower quality reduction in Lilium spp. The coat protein gene was RT-PCR-amplified from total RNA extracted from infected lily leaves and the amplified fragment was cloned into the pRSET expression vector tagged with a His-MBP. The plasmid of recombinant coat protein was used to transform an Escherichia coli strain pLysS and was expressed. The coat protein was purified by affinity chromatography using a Ni-NTA resin. The identity of the purified protein was confirmed by SDS-PAGE. The in vitro-expressed protein was used for immunization of mice. The polyclonal and monoclonal antibodies reacted specifically for the detection of LMoV in lily extracts in Western blot. Moreover the monoclonal antibodies reacted with lily extracts in DAS-ELISA with no unspecific or heterologous reactions against other non-serologically related viruses, but the polyclonal antibodies revealed a weak reaction against both infected lily and healthy control.

• The Plant Pathology Journal
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• v.29 no.3
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• pp.338-343
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• 2013

A detection system based on a multiplex reverse transcription (RT) polymerase chain reaction (PCR) was developed to simultaneously identify multiple viruses in the lily plant. The most common viruses infecting lily plants are the cucumber mosaic virus (CMV), lily mottle virus (LMoV), lily symptomless virus (LSV). Leaf samples were collected at lily-cultivation facilities located in the Kangwon province of Korea and used to evaluate the detection system. Simplex and multiplex RT-PCR were performed using virus-specific primers to detect single- or mixed viral infections in lily plants. Our results demonstrate the selective detection of 3 different viruses (CMV, LMoV and LSV) by using specific primers as well as the potential of simultaneously detecting 2 or 3 different viruses in lily plants with mixed infections. Three sets of primers for each target virus, and one set of internal control primers were used to evaluate the detection system for efficiency, reliability, and reproducibility.

• Research in Plant Disease
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• v.7 no.2
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• pp.80-85
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• 2001

This study was carried out to examine tne incidences of virus diseases in lily plants cultivated in highland areas, and to develop an effective detection method. Viral symptoms on lilies in the highland areas were differentiated into mosaic, crinkle, mottle, stripe and line pattern. The distribution of symptoms on infected plants was 43.8% of mosaic, 29.2% of crinkle, and 10.9% of mottle symptoms. Six viruses such as Lily symptomless vires(LSV), Cucumber mosaic virus (CMV), Lily mottle virus (LMoV), Lily virus X (LVX, Potexvirus), Tabacco mosaic virus (TMV,Tobamovirus), and Tabacco rattle virus (TRV,Tobravirus) were detected from the infected lilies. Infection rate of Lilium oriental (cvs. Casablanca and Marcopolo) was 2~4 times higher than that of L. asiatic (cvs. Solemio and Prato). Virus detection on lilies by one-step RT-PCR (by using reverse transcription and polymerase chain reaction simultaneously) was more rapid rapid and reliable than by the conventional RT-PCR method.