Publications by authors named "Hong-Soo Choi"

62 Publications

Adaptation and Codon-Usage Preference of Apple and Pear-Infecting Apple Stem Grooving Viruses.

Microorganisms 2021 May 21;9(6). Epub 2021 May 21.

Department of Integrative Biotechnology, Sungkyunkwan University, Suwon 16419, Korea.

Apple stem grooving virus (ASGV; genus ) is an economically important virus. It has an approx. 6.5 kb, monopartite, linear, positive-sense, single-stranded RNA genome. The present study includes identification of 24 isolates-13 isolates from apple ( L.) and 11 isolates from pear ( L.)-from different agricultural fields in South Korea. The coat protein (CP) gene of the corresponding 23 isolates were amplified, sequenced, and analyzed. The CP sequences showed phylogenetic separation based on their host species, and not on the geography, indicating host adaptation. Further analysis showed that the ASGV isolated in this study followed host adaptation influenced and preferred by the host codon-usage.
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http://dx.doi.org/10.3390/microorganisms9061111DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8223792PMC
May 2021

First report of in in Korea.

Plant Dis 2021 Mar 9. Epub 2021 Mar 9.

Chungbuk National University, 34933, College of Agriculture, Life and Environment Sciences, 1 Chungdae-ro, Seowon-gu, Cheongju, S20, 412, Cheongju, Chungcheongbuk-do, Korea (the Republic of), 28644;

Brugmansia suaveolens, known as angel's trumpet, is a perennial ornamental shrub in the Solanaceae with large fragrant flowers. In June 2018, a leaf sample of B. suaveolens that showed virus-like symptoms including chlorotic spots, yellowing and mottle on leaves was collected from a greenhouse in Seongnam, South Korea for disease diagnosis (Supplementary Figure S1a, b). Disease incidence in the greenhouse was greater than 80% for about 2,000 B. suaveolens plants. To identify a causal virus, transmission electron microscopy (TEM) was used to analyze symptomatic leaf samples using leaf dips and thin section methods. Filamentous virus particles and pinwheel structures were observed, indicating the presence of a potyvirus (Supplementary Figure S1c, d). To confirm the TEM results, a symptomatic leaf sample was further analyzed by reverse-transcription polymerase chain reaction (RT-PCR) using species-specific detection primers for three potyviruses that infect Brugmansia spp.: Colombian datura virus (CDV), Brugmansia mosaic virus (BruMV), and Brugmansia suaveolens mottle virus (BsMoV) (Lucinda et al, 2008; Park et al., 2014; Verma et al., 2014). The sample was positive only for CDV. CDV is transmitted by aphids in a nonpersistent manner and mechanical inoculation and can infect plants in the Solanaceae family including tomato and tobacco (Kahn and Bartels 1968; Schubert et al. 2006; Verhoeven et al. 1996) and has been designated a quarantine virus in Korea. Additional analysis of 13 symptomatic B. suaveolens plants from the infected greenhouse found that all samples except one were infected with CDV. To isolate CDV from B. suaveolens, leaf extracts from symptomatic samples were mechanically inoculated on an assay host, Nicotiana tabacum cv. BY via three single-lesion passages followed by propagation in N. benthamiana. For the bioassay of the CDV isolate (CDV-AT-Kr), sap from infected N. benthamiana was mechanically inoculated on 31 indicator plants, including B. suaveolens (Supplementary Table S2). CDV-AT-Kr induced chlorotic local lesions, necrotic local lesions, mottle, and/or mosaic systemically in 10 Nicotiana spp., and mottle and yellowing in tomato. On inoculated B. suaveolens, te mild mottle symptom was reproduced. No symptoms were observed in pepper or Datura stramonium. These results were confirmed by RT-PCR. To characterize CDV-AT-Kr genetically, the complete genome sequence of CDV-AT-Kr was obtained by RT-PCR using specific primers (Supplementary Table S3) and deposited in GenBank (accession no. MW075268). The CDV-AT-Kr RNA consists of 9,620 nt, encoding a polyprotein of 3,076 aa. BLASTn analysis showed that CDV-AT had maximum nucleotide identities of 98.9% at the complete genome level with a CDV isolate (accession no. JQ801448) from N. tabacum in the UK. To our knowledge, this is the first report of CDV infection in B. suaveolens in Korea and the second report in the world of the complete genome sequence. As B. suaveolens is cultivated by vegetative propagation, production and maintenance of virus-free, healthy B. suaveolens is needed. In addition, as new CDV hosts have been repeatedly reported (Pacifico et al., 2016; Salamon et al., 2015; Tomitaka et al., 2014; Verma et al., 2014), we are monitoring nationwide occurrence to prevent the spread of the virus to other crops.
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http://dx.doi.org/10.1094/PDIS-12-20-2675-PDNDOI Listing
March 2021

First report of cucurbit chlorotic yellows virus infecting cucumber in South Korea.

Plant Dis 2021 Jan 6. Epub 2021 Jan 6.

Chungbuk National University, 34933, College of Agriculture, Life and Environment Sciences, Cheongju, Chungcheongbuk-do, Korea (the Republic of);

In October 2018, cucumber plants showing yellowing and chlorotic mottle symptoms were observed in a greenhouse in Chungbuk, South Korea. The observed symptoms were similar to those caused by cucurbit aphid-borne yellows virus (CABYV), which has been detected on cucumber plants in the region since it was reported on melon in Korea in 2015 (Lee et al 2015). To identify the potential agents causing these symptoms, 28 samples from symptomatic leaves and fruit of cucumber plants were subjected to total RNA extraction using the Plant RNA Prep Kit (Biocubesystem, Korea). Reverse transcription polymerase chain (RT-PCR) was performed on total RNA using CABYV specific primers and protocols (Kwak et al. 2018). CABYV was detected in 17 of the 28 samples, while 11 symptomatic samples tested negative. In order to identify the cause of the symptoms, RT-PCR was performed using cucurbit chlorotic yellows virus (CCYV) and cucurbit yellow stunting disorder virus (CYSDV) specific primers (Wintermantel et al. 2019). Eight of the 28 samples were positive using the CCYV specific primers while seven samples were infected with only CCYV and one contained a mixed infection of CABYV with CCYV. None of the samples tested positive for CYSDV. The expected 373 nt amplicons of CCYV were bi-directionally sequenced, and BLASTn analysis showed that the nucleotide sequences shared 98 to 100% identity with CCYV isolates from East Asia, including NC0180174 from Japan. Two pairs of primers for amplification of the complete coat protein and RNA-dependent RNA polymerase (RdRp) genes (Wintermantel et al., 2019) were used to amplify the 753bp coat protein and 1517bp RdRp genes, respectively. Amplicons of the expected sizes were obtained from a CCYV single infection and ligated into the pGEM T- Easy vector (Promega, WI, USA). Three clones from each amplicon were sequenced and aligned using Geneious Prime and found to have identical sequences (Genbank accession nos. MW033300, MW033301). The CP and RdRp sequences demonstrated 99% nucleotide and 100% amino acid identity with the respective genes and proteins of the CCYV isolates from Japan. This study documents the first report of CCYV in Korea. Since CCYV was first detected on melon in Japan, it has been reported in many other countries including those in East Asia, the Middle East, Southern Europe, North Africa, and recently in North America. CCYV has the potential to become a serious threat to production of cucurbit crops in Korea, particularly due to the increasing prevalence of the whitefly, , in greenhouse production systems. It will be important to continue monitoring for CCYV and determine potential alternate hosts in the region to manage and prevent further spread of CCYV in Korea.
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http://dx.doi.org/10.1094/PDIS-10-20-2254-PDNDOI Listing
January 2021

First report of Tomato spotted wilt virus in in Korea.

Plant Dis 2020 Dec 17. Epub 2020 Dec 17.

Rural Development Administration, 54670, National Institute of Agricultural Sciences, Crop Protection, Jeonju, Jeollabuk-do, Korea (the Republic of);

Butterbur ( [Siebold & Zucc.] Maxim.) is a perennial herb of the Asteraceae family that is cultivated for medicinal and nutritional purposes. Due to long-term vegetative propagation of virus-infected native species, the yield and quality of butterbur plants have deteriorated. Five viruses have been reported to infect this species: alfalfa mosaic virus (AMV), arabis mosaic virus (ArMV), butterbur mosaic virus (ButMV), broad bean wilt virus 2 (BBWV-2), and cucumber mosaic virus (CMV) (Ham et al. 2016; Tochihara and Tamura 1976). From 2018 to 2019, butterbur plants in four greenhouses in Nonsan, South Korea (Supplementary Figure S1a, b) were found to show virus-like symptoms such as chlorotic and necrotic ring spots, necrosis, and mild mosaic on the leaves. Disease incidence was greater than 80% in one greenhouse (~1,000 m2). To identify the causal virus, we collected 17 symptomatic butterbur leaf samples from these greenhouses and performed reverse-transcription polymerase chain reaction (RT-PCR) analysis using species-specific detection primers for the five reported viruses and tomato spotted wilt virus (TSWV) (Supplementary Table S2). RT-PCR results showed that 12 samples from three greenhouses showing necrotic ring spots and mosaic symptoms were infected with a mixture of TSWV and ButMV, whereas 5 samples from one greenhouse showing mild mosaic symptoms were infected only with ButMV. TSWV (genus , family ) is transmitted by thrips and causes serious damage to a wide range of economically important plants (Pappu et al. 2009). ButMV (genus , family ) is transmitted by aphids, as well as infected vegetative propagation material (Hashimoto et al. 2009) and is the most predominant virus in butterbur in Korea (Ham et al. 2016). To isolate TSWV from butterbur, leaf extracts from symptomatic samples were mechanically inoculated on an assay host, , via three single-lesion passages followed by propagation in cv. Samsun. Thirty different indicator plant species were used for the bioassay of the TSWV isolate (TSWV-NS-BB20) by mechanical inoculation method (Supplementary Table S3). RT-PCR analysis confirmed that TSWV-NS-BB20 induced necrotic local lesions and mosaic on species and ring spots and mosaic on tomatoes and peppers. Notably, TSWV-NS-BB20 reproduced necrotic local lesions and mild mosaic symptoms on butterbur plants which were infected with ButMV with no obvious symptoms. To characterize TSWV-NS-BB20 genetically, the complete genome sequences of L (8914 nt), M (4751 nt), and S (2917 nt) RNA segments were obtained by RT-PCR using specific primers for TSWV as described previously (Kwak et al., 2020). The obtained sequences were deposited in GenBank under accession nos. MT643236, MT842841, and MN854654, respectively. BLASTn analysis showed that sequences of each segment had maximum nucleotide identities of 99.0, 98.9, and 98.6% to TSWV-L, M, and S (KP008128, FM163373, and KP008129) of TSWV-LL-N.05 isolate from tomato in Spain. Since 2018, TSWV outbreaks on butterbur are observed every year and thus may act as a potential source of TSWV infection for other crops of importance to Korea, such as pepper. Owing to the butterbur vegetative propagation, the identification of TSWV infection in butterbur will be helpful for future virus management to generate virus-free materials. To our knowledge, this is the first report of TSWV infection of butterbur.
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http://dx.doi.org/10.1094/PDIS-09-20-2027-PDNDOI Listing
December 2020

Identification of Viruses and Viroids Infecting Tomato and Pepper Plants in Vietnam by Metatranscriptomics.

Int J Mol Sci 2020 Oct 13;21(20). Epub 2020 Oct 13.

Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Korea.

Tomato ( L.) and pepper ( L.) plants belonging to the family are cultivated worldwide. The rapid development of next-generation sequencing (NGS) technology facilitates the identification of viruses and viroids infecting plants. In this study, we carried out metatranscriptomics using RNA sequencing followed by bioinformatics analyses to identify viruses and viroids infecting tomato and pepper plants in Vietnam. We prepared a total of 16 libraries, including eight tomato and eight pepper libraries derived from different geographical regions in Vietnam. We identified a total of 602 virus-associated contigs, which were assigned to 18 different virus species belonging to nine different viral genera. We identified 13 different viruses and two viroids infecting tomato plants and 12 viruses and two viroids infecting pepper plants with viruses as dominantly observed pathogens. Our results showed that multiple infection of different viral pathogens was common in both plants. Moreover, geographical region and host plant were two major factors to determine viral populations. Taken together, our results provide the comprehensive overview of viral pathogens infecting two important plants in the family grown in Vietnam.
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http://dx.doi.org/10.3390/ijms21207565DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7593927PMC
October 2020

Development of novel detection system for sweet potato leaf curl virus using recombinant scFv.

Sci Rep 2020 05 15;10(1):8039. Epub 2020 May 15.

College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon, 16419, Republic of Korea.

Sweet potato leaf curl virus (SPLCV) causes yield losses in sweet potato cultivation. Diagnostic techniques such as serological detection have been developed because these plant viruses are difficult to treat. Serological assays have been used extensively with recombinant antibodies such as whole immunoglobulin or single-chain variable fragments (scFv). An scFv consists of variable heavy (V) and variable light (V) chains joined with a short, flexible peptide linker. An scFv can serve as a diagnostic application using various combinations of variable chains. Two SPLCV-specific scFv clones, F7 and G7, were screened by bio-panning process with a yeast cell which expressed coat protein (CP) of SPLCV. The scFv genes were subcloned and expressed in Escherichia coli. The binding affinity and characteristics of the expressed proteins were confirmed by enzyme-linked immunosorbent assay using SPLCV-infected plant leaves. Virus-specific scFv selection by a combination of yeast-surface display and scFv-phage display can be applied to detection of any virus.
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http://dx.doi.org/10.1038/s41598-020-64996-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7228925PMC
May 2020

Life Cycle-Based Host Range Analysis for Tomato Spotted Wilt Virus in Korea.

Plant Pathol J 2020 Feb 1;36(1):67-75. Epub 2020 Feb 1.

Highland Agriculture Research Institute, National Institute of Crop Science, Rural Development Administration, Pyeongchang 25342, Korea.

(TSWV) is one of the plant viruses transmitted by thrips and causes severe economic damage to various crops. From 2008 to 2011, to identify natural host species of TSWV in South Korea, weeds and crops were collected from 5 regions (Seosan, Yesan, Yeonggwang, Naju, and Suncheon) where TSWV occurred and were identified as 1,104 samples that belong to 144 species from 40 families. According to reverse transcription-polymerase chain reaction, TSWV was detected from 73 samples from 23 crop species, 5 of which belonged to family Solanaceae. Additionally, 42 weed species were confirmed as natural hosts of TSWV with three different life cycles, indicating that these weed species could play an important role as virus reservoirs during no cultivation periods of crops. This study provides up-to-date comprehensive information for TSWV natural hosts in South Korea.
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http://dx.doi.org/10.5423/PPJ.FT.12.2019.0290DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7012576PMC
February 2020

Complete Genome Sequences and Evolutionary Analysis of Isolates from Melon in Korea.

Plant Pathol J 2018 Dec 1;34(6):532-543. Epub 2018 Dec 1.

Crop Protection Division, National Institute of Agricultural Science, Wanju 55365, Korea.

Complete genome sequences of 22 isolates of (CABYV), collected from melon plants showing yellowing symptom in Korea during the years 2013-2014, were determined and compared with previously reported CABYV genome sequences. The complete genomes were found to be 5,680-5,684 nucleotides in length and to encode six open reading frames (ORFs) that are separated into two regions by a non-coding internal region (IR) of 199 nucleotides. Their genomic organization is typical of the genus . Based on phylogenetic analyses of complete nucleotide (nt) sequences, CABYV isolates were divided into four groups: Asian, Mediterranean, Taiwanese, and R groups. The Korean CABYV isolates clustered with the Asian group with > 94% nt sequence identity. In contrast, the Korean CABYV isolates shared 87-89% sequence identities with the Mediterranean group, 88% with the Taiwanese group, 81-84% with the CABYV-R group, and 72% with another , . Recombination analyses identified 24 recombination events (12 different recombination types) in the analyzed CABYV population. In the Korean CABYV isolates, four recombination types were detected from eight isolates. Two recombination types were detected in the IR and P3-P5 regions, respectively, which have been reported as hotspots for recombination of CABYV. This result suggests that recombination is an important evolutionary force in the genetic diversification of CABYV populations.
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http://dx.doi.org/10.5423/PPJ.OA.03.2018.0049DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6305177PMC
December 2018

Complete Genome Sequence Analysis of Two Divergent Groups of Isolates Collected from Korea.

Plant Pathol J 2018 Oct 1;34(5):451-457. Epub 2018 Oct 1.

Crop Protection Division, National Institute of Agricultural Science, Wanju 55365, Korea.

The (SPCFV), of the genus (family Betaflexiviridae), was first detected as one of several viruses infecting sweet potatoes ( L.) in Korea. Out of 154 sweet potato samples collected in 2012 that were showing virus-like symptoms, 47 (31%) were infected with SPCFV, along with other viruses. The complete genome sequences of four SPCFV isolates were determined and analyzed using previously reported genome sequences. The complete genomes were found to contain 9,104-9,108 nucleotides, excluding the poly-A tail, containing six putative open reading frames (ORFs). Further, the SPCFV Korean isolates were divided into two groups (Group I and Group II) by phylogenetic analysis based on the complete nucleotide sequences; Group I and Group II had low nucleotide sequence identities of about 73%. For the first time, we determined the complete genome sequence for the Group II SPCFV isolates. The amino acid sequence identity in coat proteins (CP) between the two groups was over 90%, whereas the amino acid sequence identity in other proteins was less than 80%. In addition, SPCFV Korean isolates had a low amino acid sequence identity (61% CPs and 47% in the nucleotide- binding protein [NaBp] region) to that of (MYaV), a typical .
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http://dx.doi.org/10.5423/PPJ.NT.03.2018.0042DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6200045PMC
October 2018

Phylogeographic analysis of the full genome of Sweepovirus to trace virus dispersal and introduction to Korea.

PLoS One 2018 13;13(8):e0202174. Epub 2018 Aug 13.

Department of Genetic Engineering, Sungkyunkwan University, Suwon, Korea.

Sweet potato is a vegetatively propagated crop that is produced for both growth in Korean fields and for export out of the country. The viruses that are present in introduced sweet potatoes can spread both domestically and to foreign countries. Determining the time and path of virus movement could help curtail its spread and prevent future dispersal of related viruses. Determining the consequences of past virus and sweet potato dispersal could provide insight into the ecological and economic risks associated with other sweet potato-infecting viral invasions. We therefore applied Bayesian phylogeographic inferences and recombination analyses of the available Sweepovirus sequences (including 25 Korean Sweepovirus genomes) and reconstructed a plausible history of Sweepovirus diversification and movement across the globe. The Mediterranean basin and Central America were found to be the launchpad of global Sweepovirus dispersal. Currently, China and Brazil are acting as convergence regions for Sweepoviruses. Recently reported Korean Sweepovirus isolates were introduced from China in a recent phase and the regions around China and Brazil continue to act as centers of Sweepovirus diversity and sites of ongoing Sweepovirus evolution. The evidence indicates that the region is an epidemiological hotspot, which suggests that novel Sweepovirus variants might be found.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0202174PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6089449PMC
February 2019

Phylogenetic Characterization of Population in Korea: Evidence of Reassortment between Isolates from Different Origins.

Plant Pathol J 2018 Jun 1;34(3):199-207. Epub 2018 Jun 1.

Crop Protection Division, National Academy of Agricultural Science, Rural Development Administration, Wanju 55365, Korea.

(ToCV) is a whitefly-transmitted and phloem-limited crinivirus. In 2013, severe interveinal chlorosis and bronzing on tomato leaves, known symptoms of ToCV infection, were observed in greenhouses in Korea. To identify ToCV infection in symptomatic tomato plants, RT-PCR with ToCV-specific primers was performed on leaf samples collected from 11 tomato cultivating areas where ToCV-like symptoms were observed in 2013 and 2014. About half of samples (45.18%) were confirmed as ToCV-infected, and the complete genome of 10 different isolates were characterized. This is the first report of ToCV occurring in Korea. The phylogenetic relationship and genetic variation among ToCV isolates from Korea and other countries were also analysed. When RNA1 and RNA2 are analysed separately, ToCV isolates were clustered into three groups in phylogenetic trees, and ToCV Korean isolates were confirmed to belong to two groups, which were geographically separated. These results suggested that Korean ToCV isolates originated from two independent origins. However, the RNA1 and RNA2 sequences of the Yeonggwang isolate were confirmed to belong to different groups, which indicated that ToCV RNA1 and RNA2 originated from two different origins and were reassorted in Yeonggwang, which is the intermediate point of two geographically separated groups.
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http://dx.doi.org/10.5423/PPJ.OA.10.2017.0220DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5985646PMC
June 2018

Molecular dissection of distinct symptoms induced by tomato chlorosis virus and tomato yellow leaf curl virus based on comparative transcriptome analysis.

Virology 2018 03 6;516:1-20. Epub 2018 Jan 6.

Department of International Agricultural Technology and Institutes of Green Bio Science and Technology, Seoul National University, Pyeongchang 25354, Republic of Korea. Electronic address:

The viral infection of plants may cause various physiological symptoms associated with the reprogramming of plant gene expression. However, the molecular mechanisms and associated genes underlying disease symptom development in plants infected with viruses are largely unknown. In this study, we employed RNA sequencing for in-depth molecular characterization of the transcriptional changes associated with the development of distinct symptoms induced by tomato chlorosis virus (ToCV) and tomato yellow leaf curl virus (TYLCV) in tomato. Comparative analysis of differentially expressed genes revealed that ToCV and TYLCV induced distinct transcriptional changes in tomato and resulted in the identification of important genes responsible for the development of symptoms of ToCV (i.e., chlorosis and anthocyanin accumulation) and TYLCV (i.e., yellowing, stunted growth, and leaf curl). Our comprehensive transcriptome analysis can provide molecular strategies to reduce the severity of disease symptoms as well as new insights for the development of virus-resistant crops.
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http://dx.doi.org/10.1016/j.virol.2018.01.001DOI Listing
March 2018

Virus Incidence of Sweet Potato in Korea from 2011 to 2014.

Plant Pathol J 2017 Oct 1;33(5):467-477. Epub 2017 Oct 1.

Crop Protection Division, National Academy of Agricultural Science, Wanju 55365, Korea.

A nationwide survey was performed to investigate the current incidence of viral diseases in Korean sweet potatoes for germplasm and growing fields from 2011 to 2014. A total of 83.8% of the germplasm in Korea was infected with viruses in 2011. Commercial cultivars that were used to supply growing fields were infected at a rate of 62.1% in 2012. Among surveyed viruses, the incidence of five species that infect sweet potato decreased between 2012 and 2013, and then increased again in 2014. Representatively, the incidence of Sweet potato feathery mottle virus (SPFMV) was 87.0% in 2012, 20.7% in 2013 and then increased to 35.3% in 2014. Unlike RNA viruses, DNA viruses were shown to decrease continuously. The incidence of (SPLCV) was 5.5% in 2003, 59.5% in 2011, and 47.4% in 2012. It then decreased continuously year by year to 33.2% in 2013, and then 25.6% in 2014. While the infection rate of each virus species showed a tendency to decline, the virus infection status was more variable in 2013 and 2014. Nevertheless, the high rate of single infections and mixed infection combinations were more variable than the survey results from 2012. As shown in the results from 2013, the most prevalent virus infection was a single infection at 27.6%, with the highest rate of infection belonging to sweet potato symptomless virus-1 (SPSMV-1) (12.9%). Compared to 2013, infection combinations were more varied in 2014, with a total of 122 kinds of mixed infection.
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http://dx.doi.org/10.5423/PPJ.OA.08.2016.0167DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5624489PMC
October 2017

Movement protein of broad bean wilt virus 2 serves as a determinant of symptom severity in pepper.

Virus Res 2017 10 29;242:141-145. Epub 2017 Sep 29.

Crop Protection Division, National Institute of Agricultural Sciences, Rural Development Administration, Wanju 55365, Republic of Korea.

Broad bean wilt virus 2 (BBWV2, genus Fabavirus, family Secoviridae) has a wide host range and infects many economically important crops. Various isolates of BBWV2 have been identified from diverse host plants, and their molecular and biological characteristics have been investigated. In our previous study, we demonstrated that BBWV2 RNA2 contains a symptom determinant(s) capable of enhancing symptom severity by utilizing infectious full-length cDNA clones of two distinct strains of BBWV2, pBBWV2-PAP1 (a severe strain) and pBBWV2-RP1 (a mild strain). In the present study, to identify the symptom determinant(s) of BBWV2, we exploited disease responses of pBBWV2-PAP1- and pBBWV2-RP1-derived chimeric viruses and amino acid substitution mutant viruses in Nicotiana benthamiana and pepper (Capsicum annuum Quarri) and demonstrated that the movement protein (MP) encoded in BBWV RNA2 is the determinant of disease symptom severity in both plants. A single amino acid substitution in the MP was sufficient for changing symptom severity of BBWV2. Our finding provides a role for the MP as a symptom determinant in BBWV2 and increases the understanding of the basis of molecular interactions between host plants and BBWV2.
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http://dx.doi.org/10.1016/j.virusres.2017.09.024DOI Listing
October 2017

Seed Transmission of in White Soybean ().

Plant Pathol J 2017 Aug 1;33(4):424-428. Epub 2017 Aug 1.

Department of Genetic Engineering, Sungkyunkwan University, Suwon 16419, Korea.

(TYLCV) infection of the common bean () has been reported, but soybean () has not previously been identified as a TYLCV host. Five cultivars of white soybean were agro-inoculated using an infectious TYLCV clone. At 30 days post-inoculation, they showed infection rates of 25% to 100%. Typical TYLCV symptoms were not observed in any inoculated plants. To examine whether TYLCV was transmitted in soybean seeds, DNA was isolated from bundles of five randomly selected seeds from TYLCV-inoculated soybean plants and amplified with a TYLCV-specific primer set. With the exception of one bundle, all bundles of seeds were verified to be TYLCV-infected. Virus dissemination was also confirmed in three of the 14 bunches. Viral replication was also identified in seeds and seedlings. This is the first report demonstrating that soybean is a TYLCV host, and that TYLCV is a seed-transmissible virus in white soybean.
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http://dx.doi.org/10.5423/PPJ.NT.02.2017.0043DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5538446PMC
August 2017

Erratum to: Complete genome sequence of longan witches' broom-associated virus, a novel member of the family Potyviridae.

Arch Virol 2017 09;162(9):2891

Crop Protection Division, National Institute of Agricultural Sciences, Rural Development Administration, Wanju, 55365, Republic of Korea.

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http://dx.doi.org/10.1007/s00705-017-3458-2DOI Listing
September 2017

Efficient Transmission and Propagation of Tomato Chlorosis Virus by Simple Single-Leaflet Grafting.

Plant Pathol J 2017 Jun 1;33(3):345-349. Epub 2017 Jun 1.

Department of International Agricultural Technology and Institutes of Green Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea.

Tomato chlorosis virus (ToCV), a member of the genus , has caused an epidemic disease in tomato worldwide. ToCV is phloem-limited and transmitted by whiteflies in a semi-persistent manner, but not by mechanical inoculation. Experimental propagation of ToCV has been performed primarily by using whitefly-mediated inoculation. To develop a simple and convenient method for transmission of ToCV, we investigated grafting single-leaflets from tomato plants infected with ToCV to recipient tomato seedlings. Forty-one of 46 tomato seedlings tested were grafted successfully with single-leaflets infected with ToCV. Among them, 36 seedlings (87.8%) were systemically infected with ToCV and developed typical symptoms. Our results demonstrated that single-leaflet grafting could provide a sufficient amount of inoculum for the transmission of ToCV to the grafted seedlings.
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http://dx.doi.org/10.5423/PPJ.NT.02.2017.0039DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5461053PMC
June 2017

Complete genome sequence of longan witches' broom-associated virus, a novel member of the family Potyviridae.

Arch Virol 2017 Sep 13;162(9):2885-2889. Epub 2017 May 13.

Crop Protection Division, National Institute of Agricultural Sciences, Rural Development Administration, Wanju, 55365, Republic of Korea.

The complete genome sequence of a new virus isolated from a longan (Dimocarpus longan Lour.) plant showing witches' broom syndrome was determined. The viral genome is composed of a monopartite single-stranded RNA of 9,428 nucleotides excluding the 3' poly(A) tail and contains one large single open reading frame encoding a polyprotein of 3086 amino acids. BLAST searches of protein databases showed that the encoded polyprotein has a maximum amino acid sequence identity of 35% (with 85% coverage) to that of the isolate Minnesota of rose yellow mosaic virus (RoYMV; family Potyviridae; genus not assigned). Molecular and phylogenetic analysis of the genome and encoded protein sequences showed that the identified virus has the general features that are characteristic of members of the family Potyviridae although it has extremely low sequence similarity to known members of the family Potyviridae. The name longan witches' broom-associated virus (LWBaV) is proposed for this new virus, which may be considered a member of a new genus in the family Potyviridae.
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http://dx.doi.org/10.1007/s00705-017-3405-2DOI Listing
September 2017

The complete genome sequence of a novel virus, bellflower veinal mottle virus, suggests the existence of a new genus within the family Potyviridae.

Arch Virol 2017 Aug 22;162(8):2457-2461. Epub 2017 Apr 22.

Crop Protection Division, National Institute of Agricultural Sciences, Rural Development Administration, Wanju, 55365, Republic of Korea.

A new virus was isolated from a bellflower (Campanula takesimana) plant showing veinal mottle symptoms, and its complete genome sequence was determined. The viral genome consists of a positive-sense single-stranded RNA of 8,259 ribonucleotides. Electron microscopic observation revealed that the viral genome is packaged as a filamentous particle with an average length of approximately 760 nm. BLAST searches of protein databases showed that the encoded polyprotein has a maximum amino acid sequence identity of 34.1% (with 95% coverage) to that of the isolate AD of Chinese yam necrotic mosaic virus (CYNMV; genus Macluravirus). Phylogenetic analysis and comparison of the encoded amino acid sequences with those of other viruses demonstrated that the identified virus shows minimal sequence similarity to known viruses and should therefore be considered a member of a new genus in the family Potyviridae. The name bellflower veinal mottle virus (BVMoV) is proposed for this new virus.
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http://dx.doi.org/10.1007/s00705-017-3374-5DOI Listing
August 2017

Engineering of soybean mosaic virus as a versatile tool for studying protein-protein interactions in soybean.

Sci Rep 2016 Feb 29;6:22436. Epub 2016 Feb 29.

Department of Agricultural Biotechnology and Plant Genomics and Breeding Institute, Seoul National University, Seoul 151-921, Republic of Korea.

Transient gene expression approaches are valuable tools for rapid introduction of genes of interest and characterization of their functions in plants. Although agroinfiltration is the most effectively and routinely used method for transient expression of multiple genes in various plant species, this approach has been largely unsuccessful in soybean. In this study, we engineered soybean mosaic virus (SMV) as a dual-gene delivery vector to simultaneously deliver and express two genes in soybean cells. We further show the application of the SMV-based dual vector for a bimolecular fluorescence complementation assay to visualize in vivo protein-protein interactions in soybean and for a co-immunoprecipitation assay to identify cellular proteins interacting with SMV helper component protease. This approach provides a rapid and cost-effective tool for transient introduction of multiple traits into soybean and for in vivo characterization of the soybean cellular protein interaction network.
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http://dx.doi.org/10.1038/srep22436DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4772626PMC
February 2016

Establishment of a Simple and Rapid Gene Delivery System for Cucurbits by Using Engineered of Zucchini yellow mosaic virus.

Plant Pathol J 2016 Feb 1;32(1):70-6. Epub 2016 Feb 1.

Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea; Plant Genomics and Breeding Institute, Seoul National University, Seoul 151-921, Korea.

The infectious full-length cDNA clone of zucchini yellow mosaic virus (ZYMV) isolate PA (pZYMV-PA), which was isolated from pumpkin, was constructed by utilizing viral transcription and processing signals to produce infectious in vivo transcripts. Simple rub-inoculation of plasmid DNAs of pZYMV-PA was successful to cause infection of zucchini plants (Cucurbita pepo L.). We further engineered this infectious cDNA clone of ZYMV as a viral vector for systemic expression of heterologous proteins in cucurbits. We successfully expressed two reporter genes including gfp and bar in zucchini plants by simple rub-inoculation of plasmid DNAs of the ZYMV-based expression constructs. Our method of the ZYMV-based viral vector in association with the simple rub-inoculation provides an easy and rapid approach for introduction and evaluation of heterologous genes in cucurbits.
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http://dx.doi.org/10.5423/PPJ.NT.08.2015.0173DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4755678PMC
February 2016

Identification of Leonurus sibiricus as a Weed Reservoir for Three Pepper-Infecting Viruses.

Plant Pathol J 2016 Feb 1;32(1):65-9. Epub 2016 Feb 1.

Crop Protection Division, National Academy of Agricultural Science, RDA, Wanju 565-851, Korea.

In plant virus ecology, weeds are regarded as wild reservoirs of viruses and as potential sources for insect-mediated transmission of viruses. During field surveys in 2013-2014, three Leonurus sibiricus plants showing virus-like symptoms were collected from pepper fields in Daegu, Seosan, and Danyang in Korea. Molecular diagnosis assays showed that the collected L. sibiricus samples were infected with either Tomato spotted wilt virus (TSWV), Pepper mild mottle virus (PMMoV), or Beet western yellow virus (BWYV), respectively. Since this is the first identification of TSWV, PMMoV, and BWYV from L. sibiricus, complete genome sequences of three virus isolates were determined to examine their phylogenetic relationships with the previously reported strains and isolates. Phylogenetic analyses performed using full genome sequences of the viruses showed the isolates of TSWV and PMMoV obtained from L. sibiricus are closely related to the pepper isolates of the corresponding viruses. Our results suggest that L. sibiricus could act an alternative host and reservoir of viruses that cause damages in pepper fields.
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http://dx.doi.org/10.5423/PPJ.NT.07.2015.0138DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4755677PMC
February 2016

Tomato yellow leaf curl virus (TYLCV-IL): a seed-transmissible geminivirus in tomatoes.

Sci Rep 2016 Jan 8;6:19013. Epub 2016 Jan 8.

Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Korea.

Tomato yellow leaf curl virus (TYLCV) is one of the most well-known tomato-infecting begomoviruses and transmitted by Bemisia tabaci. Seed transmission has previously been reported for some RNA viruses, but TYLCV has not previously been described as a seed-borne virus. In 2013 and 2014, without whitefly-mediated transmission, TYLCV was detected in young tomato plants germinated from fallen fruits produced from TYLCV-infected tomato plants in the previous cultivation season. In addition, TYLCV-Israel (TYLCV-IL) was also detected in seeds and their seedlings of TYLCV-infected tomato plants that were infected by both viruliferous whitefly-mediated transmission and agro-inoculation. The seed infectivity was 20-100%, respectively, and the average transmission rate to seedlings was also 84.62% and 80.77%, respectively. TYLCV-tolerant tomatoes also produced TYLCV-infected seeds, but the amount of viral genome was less than seen in TYLCV-susceptible tomato plants. When tomato plants germinated from TYLCV-infected seeds, non-viruliferous whiteflies and healthy tomato plants were placed in an insect cage together, TYLCV was detected from whiteflies as well as receiver tomato plants six weeks later. Taken together, TYLCV-IL can be transmitted via seeds, and tomato plants germinated from TYLCV-infected seeds can be an inoculum source of TYLCV. This is the first report about TYLCV seed transmission in tomato.
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http://dx.doi.org/10.1038/srep19013DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4705557PMC
January 2016

Establishment of an Agrobacterium-mediated Inoculation System for Cucumber Green Mottle Mosaic Virus.

Plant Pathol J 2015 Dec 30;31(4):433-7. Epub 2015 Dec 30.

Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea ; Plant Genomics and Breeding Institute, Seoul National University, Seoul 151-921, Korea.

The infectious full-length cDNA clones of Cucumber green mottle mosaic virus (CGMMV) isolates KW and KOM, which were isolated from watermelon and oriental melon, respectively, were constructed under the control of the cauliflower mosaic virus 35S promoter. We successfully inoculated Nicotiana benthamiana with the cloned CGMMV isolates KW and KOM by Agrobacterium-mediated infiltration. Virulence and symptomatic characteristics of the cloned CGMMV isolates KW and KOM were tested on several indicator plants. No obvious differences between two cloned isolates in disease development were observed on the tested indicator plants. We also determined full genome sequences of the cloned CGMMV isolates KW and KOM. Sequence comparison revealed that only four amino acids (at positions 228, 699, 1212, and 1238 of the replicase protein region) differ between the cloned isolates KW and KOM. A previous study reported that the isolate KOM could not infect Chenopodium amaranticolor, but the cloned KOM induced chlorotic spots on the inoculated leaves. When compared with the previously reported sequence of the original KOM isolate, the cloned KOM contained one amino acid mutation (Ala to Thr) at position 228 of the replicase protein, suggesting that this mutation might be responsible for induction of chlorotic spots on the inoculated leaves of C. amaranticolor.
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http://dx.doi.org/10.5423/PPJ.NT.06.2015.0123DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4677753PMC
December 2015

Molecular Characterization of Five Potyviruses Infecting Korean Sweet Potatoes Based on Analyses of Complete Genome Sequences.

Plant Pathol J 2015 Dec 30;31(4):388-401. Epub 2015 Dec 30.

Crop Protection Division, National Academy of Agricultural Science, Wanju 565-851, Korea.

Sweet potatoes (Ipomea batatas L.) are grown extensively, in tropical and temperate regions, and are important food crops worldwide. In Korea, potyviruses, including Sweet potato feathery mottle virus (SPFMV), Sweet potato virus C (SPVC), Sweet potato virus G (SPVG), Sweet potato virus 2 (SPV2), and Sweet potato latent virus (SPLV), have been detected in sweet potato fields at a high (~95%) incidence. In the present work, complete genome sequences of 18 isolates, representing the five potyviruses mentioned above, were compared with previously reported genome sequences. The complete genomes consisted of 10,081 to 10,830 nucleotides, excluding the poly-A tails. Their genomic organizations were typical of the Potyvirus genus, including one target open reading frame coding for a putative polyprotein. Based on phylogenetic analyses and sequence comparisons, the Korean SPFMV isolates belonged to the strains RC and O with >98% nucleotide sequence identity. Korean SPVC isolates had 99% identity to the Japanese isolate SPVC-Bungo and 70% identity to the SPFMV isolates. The Korean SPVG isolates showed 99% identity to the three previously reported SPVG isolates. Korean SPV2 isolates had 97% identity to the SPV2 GWB-2 isolate from the USA. Korean SPLV isolates had a relatively low (88%) nucleotide sequence identity with the Taiwanese SPLV-TW isolates, and they were phylogenetically distantly related to SPFMV isolates. Recombination analysis revealed that possible recombination events occurred in the P1, HC-Pro and NIa-NIb regions of SPFMV and SPLV isolates and these regions were identified as hotspots for recombination in the sweet potato potyviruses.
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http://dx.doi.org/10.5423/PPJ.OA.04.2015.0072DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4677748PMC
December 2015

Characterization of Melon necrotic spot virus Occurring on Watermelon in Korea.

Plant Pathol J 2015 Dec 30;31(4):379-87. Epub 2015 Dec 30.

Department of Crop Protection, National Academy of Agricultural Science, RDA, Wanju 565-851, Korea.

Melon necrotic spot virus (MNSV) was recently identified on watermelon (Citrullus vulgaris) in Korea, displaying as large necrotic spots and vein necrosis on the leaves and stems. The average occurrence of MNSV on watermelon was found to be 30-65% in Hapcheon and Andong City, respectively. Four isolates of the virus (MNSV-HW, MNSV-AW, MNSV-YW, and MNSV-SW) obtained from watermelon plants in different areas were non-pathogenic on ten general indicator plants, including Chenopodium quinoa, while they infected systemically six varieties of Cucurbitaceae. The virus particles purified by 10-40% sucrose density gradient centrifugation had a typical ultraviolet spectrum, with a minimum at 245 nm and a maximum at 260 nm. The morphology of the virus was spherical with a diameter of 28-30 nm. Virus particles were observed scattered throughout the cytoplasm of watermelon cells, but no crystals were detected. An ELISA was conducted using antiserum against MNSV-HW; the optimum concentrations of IgG and conjugated IgG for the assay were 1 μl/ml and a 1:8,000-1:10,000 dilutions, respectively. Antiserum against MNSV-HW could capture specifically both MNSV-MN from melon and MNSV-HW from watermelon by IC/RT-PCR, and they were effectively detected with the same specific primer to produce product of 1,172 bp. The dsRNA of MNSV-HW had the same profile (4.5, 1.8, and 1.6 kb) as that of MNSV-MN from melon. The nucleotide sequence of the coat protein of MNSV-HW gave a different phylogenetic tree, having 17.2% difference in nucleotide sequence compared with MNSV isolates from melon.
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http://dx.doi.org/10.5423/PPJ.OA.11.2014.0124DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4677747PMC
December 2015

A determinant of disease symptom severity is located in RNA2 of broad bean wilt virus 2.

Virus Res 2016 Jan 30;211:25-8. Epub 2015 Sep 30.

Crop Protection Division, National Academy of Agricultural Science, Rural Development Administration, Suwon 441-707, Republic of Korea. Electronic address:

Broad bean wilt virus 2 (BBWV2), which belongs to the genus Fabavirus, is a destructive pathogen of many economically important horticultural and ornamental crops. In this study, we constructed infectious full-length cDNA clones of two distinct isolates of BBWV2 under control of the cauliflower mosaic virus 35S promoter. BBWV2-PAP1 isolated from paprika (Capsicum annuum var. gulosum) induces severe disease symptoms in various pepper varieties, whereas BBWV2-RP1 isolated from red pepper (Capsicum annuum L.) causes mild symptoms. Agrobacterium-mediated inoculation of the infectious cDNA clones of BBWV2-PAP1 and RP1 resulted in the same symptoms as the original virus isolates. The infectious cDNA clones of BBWV2-PAP1 and RP1 were used to examine the symptoms induced by pseudorecombinants between the two isolates to localize in which of the two genomic RNAs are the symptom severity determinants in BBWV2. The pseudorecombinant of RP1-RNA1 and PAP1-RNA2 induced severe symptoms, similar to those caused by the parental isolate PAP1, whereas the pseudorecombinant of PAP1-RNA1 and RP1-RNA2 induced mild symptoms, similar to those caused by the parental isolate RP1. Our results suggest that BBWV2 RNA2 contains a symptom determinant(s) capable of enhancing symptom severity.
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http://dx.doi.org/10.1016/j.virusres.2015.09.018DOI Listing
January 2016

Complete genome sequence of bellflower vein chlorosis virus, a novel putative member of the genus Waikavirus.

Arch Virol 2015 Dec 14;160(12):3139-42. Epub 2015 Sep 14.

Crop Protection Division, National Academy of Agricultural Science, Rural Development Administration, Wanju, 565-851, Republic of Korea.

The complete genome sequence of a new virus isolated from a bellflower (Campanula takesimana) plant was determined. The genome of this virus is composed of monopartite single-stranded RNA of 11,649 nucleotides in length. BLAST searches of protein databases showed that the encoded polyprotein has a maximum amino acid sequence identity of 42% (with 99% coverage) to the polyprotein of the isolate Orissa of rice tungro spherical virus (RTSV; genus Waikavirus). Phylogenetic analysis strongly supports that the identified virus is a member of a new species of the genus Waikavirus. The name bellflower vein chlorosis virus (BVCV) is proposed for this new virus.
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http://dx.doi.org/10.1007/s00705-015-2606-9DOI Listing
December 2015

Tomato yellow leaf curl virus Can Overwinter in Stellaria aquatica, a Winter-Hardy TYLCV-Reservoir Weed.

Plant Dis 2015 May 30;99(5):588-592. Epub 2015 Apr 30.

Department of Genetic Engineering, Sungkyunkwan University.

Tomato yellow leaf curl virus (TYLCV), one of the most serious plant viruses in tropical and subtropical regions, is transmitted to host plants by the vector insect Bemisia tabaci. In order to control TYLCV, it is important to identify weed hosts for overwintering TYLCV. Stellaria aquatica, a winter-hardy weed, was found growing with TYLCV-infected tomato plants in greenhouse production. TYLCV was detected in S. aquatica plants by polymerase chain reaction and Southern blot hybridization analysis. The intergenic region nucleotide sequences amplified from TYLCV-infected tomato plants, TYLCV-viruliferous whiteflies, and S. aquatica were identical. During winter (December to February), TYLCV-viruliferous whiteflies and TYLCV-infected tomato plants were removed or absent from greenhouses. However, S. aquatica plants were observed over a period of 10 months from August to May in such greenhouses, and TYLCV was consistently detected in some of these plants. To investigate the transmission of TYLCV from TYLCV-infected S. aquatica plants to healthy tomato plants by whiteflies, TYLCV-infected S. aquatica plants were transplanted to pots in cages with nonviruliferous whiteflies and healthy tomato plants. After 4 weeks, tomato plants developed typical TYLCV disease symptoms, and TYLCV was detected in both whiteflies and tomato plants. These results show that S. aquatica can act as a winter-hardy reservoir for TYLCV, and suggest that this weed could play an important role in overwintering of TYLCV in tomato greenhouses.
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http://dx.doi.org/10.1094/PDIS-04-14-0352-REDOI Listing
May 2015

Complete genome sequence of yacon necrotic mottle virus, a novel putative member of the genus Badnavirus.

Arch Virol 2015 Apr 4;160(4):1139-42. Epub 2015 Feb 4.

Crop Protection Division, National Academy of Agricultural Science, Rural Development Administration, Suwon, 441-707, Republic of Korea.

The complete genome sequence of a previously undescribed virus isolated from a yacon plant exhibiting necrotic mottle, chlorosis, stunting, and leaf malformation symptoms in Gyeongju, Korea, was determined. The genome of this virus consists of one circular double-stranded DNA of 7661 bp in size. The genome contained four open reading frames (ORFs 1 to 4) on the plus strand that potentially encode proteins of 26, 32, 234, and 25 kDa. Protein BLAST analysis showed that ORF3, which is the largest ORF, has 45 % amino acid sequence identity (with 89 % coverage) to the ORF3 of fig badnavirus 1 (FBV-1), a recently identified badnavirus. Phylogenetic analysis provided further evidence that the virus identified in this study is probably a member of a new species in the genus Badnavirus. The name yacon necrotic mottle virus (YNMoV) is proposed for this new virus.
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http://dx.doi.org/10.1007/s00705-015-2341-2DOI Listing
April 2015
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