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We determined the complete genome sequence of the citrus vein enation virus (CVEV) collected from a Korean yuja tree ( Citrus junos ). The CVEV genome has 5,983 nucleotides, showing 97.3 to 98.6% identity with complete genomic sequences of other CVEV isolates, with the highest similarity being to the isolate PCJ.

Mume virus A (MuVA) of the genus

A new member of the genus Closterovirus was detected in Platycodon grandiflorus using high-throughput RNA sequencing analysis. The complete genome sequence of this new virus isolate, tentatively named “platycodon closterovirus 1” (PlaCV1), comprises 16,771 nucleotides with nine predicted open reading frames (ORFs) having the typical closterovirus genome organization. PlaCV1 shares 37%–50% nucleotide sequence identity with other known closterovirus genome sequences. The putative RNA-dependent RNA polymerase (RdRp), heat shock protein 70-like protein (HSP70h), viral heat shock protein 90-like protein (HSP90h), minor coat protein (CPm), and coat protein (CP) show 47–68%, 39–66%, 24–52%, 21–57%, and 16–35% amino acid sequence identity, respectively, to homologous proteins in previously identified closteroviruses, suggesting that it represents a distinct, new species in the genus. Phylogenetic analysis of HSP70h sequences places PlaCV1 alongside other members of the genus Closterovirus in the family Closteroviridae. To our knowledge, this study is the first report of the complete genome sequence of PlaCV1 infecting P. grandiflorus in the Republic of Korea.

The complete genome sequence of a new polerovirus found naturally infecting Artemisia princeps, artemisia virus B (ArtVB), was determined using high-throughput sequencing. The ArtVB genome comprises 6,141 nucleotides and contains six putative open reading frames (ORF0 to ORF5) with a genome structure typical of poleroviruses. A multiple sequence alignment showed that the complete ArtVB genome shares 50.98% nucleotide sequence identity with ixeridium yellow mottle virus 1 (IxYMaV-1, GenBank accession no. KT868949). ArtVB shares the highest amino acid sequence identity in P0 and P3–P5 (21.54%–51.69%) with other known poleroviruses. Phylogenetic analysis indicated that ArtVB should be considered a member of a new species within the genus Polerovirus, family Luteoviridae.

The complete genomic nucleotide sequence of hemisteptia virus A (HemVA) from a Hemisteptia lyrata Bunge plant in South Korea was identified by high-throughput sequencing. The HemVA genome consists of 6,122 nucleotides and contains seven putative open reading frames, ORF0-5 and ORF3a, encoding the putative proteins P0-P5 and P3a, respectively. Pairwise amino acid sequence analysis shows that the HemVA P1-P5 proteins have the highest sequence identity (23.68%-54.15%) to the corresponding proteins of members of the families Solemoviridae and Tombusviridae. Phylogenetic analysis of the P1-P2 and P3 amino acid sequences indicated that HemVA should be classified as a member of a distinct species in the genus Polerovirus.

We identified a tentative novel positive-strand RNA virus from Rudbeckia sp., namely, Rudbeckia citrivirus A (RuCVA). The complete genome sequence of the novel virus was 8821 nucleotides in length, excluding the poly(A) tail. It has three open reading frames (ORFs): a putative polyprotein, a movement protein, and a coat protein. Phylogenetic analysis showed that the virus was more closely related to Citrus leaf blotch virus isolates and unassigned citriviruses. The sequence identity of the virus with other citriviruses was lower than 56.9% at the complete nucleotide sequence level. For each ORF, the sequence identity was lower than 64.2% at the nucleotide level and 67.8% at the amino acid level. These results satisfied the species demarcation criteria for Betaflexiviridae. Therefore, we suggest that RuCVA is a novel member of the genus Citrivirus.

Raphanus sativus cryptic virus 2 (RsCV-2), an unclassified species belonging to the family Partitiviridae, was identified in cabbage seeds (Brassica rapa subsp. pekinensis) imported from Italy using high-throughput paired-end RNA sequencing. Three contigs derived from RsCV-2 isolate PQ shared nucleotide sequence homologies (approximately 94, 87, and 86%) with dsRNA1, 2, and 3 of RsCV-2, respectively. Phylogenetic analysis and pairwise comparison showed that RsCV-2 isolates were more closely related to deltapartitiviruses. To the best of our knowledge, this is the first report of RsCV-2 on B. rapa subsp. pekinensis in the world. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13337-021-00744-w.

The complete genomic sequence of achyranthes virus A (AcVA), from an Achyranthes bidentata Blume plant in South Korea, was determined. The genomic RNA has 9491 nucleotides (nt), excluding the 3'-terminal poly(A) tail and contains an open reading frame typical of members of the genus Potyvirus, family Potyviridae, encoding a large putative polyprotein of 3103 amino acids (aa). Pairwise comparisons showed that the AcVA sequence shares 47.81-57.78% nt sequence identity at the complete genome level, 41.89-56.41% aa sequence identity at the polyprotein level, and 50-63.8% aa sequence identity at the coat protein level with other members of genus Potyvirus. These pairwise comparison values are below the species demarcation cutoff for the family Potyviridae. Our results therefore suggest that this virus should be regarded as a novel member of the genus Potyvirus, tentatively named "achyranthes virus A".

Oral healthcare is a critical condition for high quality of life. According to statistics, gingivitis is one of major diseases in the world (The Global Burden of Disease Study 2019, 3.5 billion population worldwide). Oral diseases, that initiated by a group of multispecies bacteria with diverse extracellular slime that called biofilms, are modifiable risk factors with the cancer, cardiovascular disease, chronic respiratory disease. Hence, effective management of biofilms is important. Since biofilms are comprised of extracellular matrix that inhibit drug diffusion, the traditional antibiotic treatment requires 500 -5000 times more concentration than planktonic bacterial infection. The biofilm cleaning in dentistry includes time-consuming and painful physical brushing. In this work, we have developed a new electric toothbrush in emphasis on demonstration of convenient and painless gingivitis improvement device. For the first time, the toothbrush has been integrated with an electromagnetic technology for effective biofilm removal, known for bioelectric effect (BE). Biofilms consists of electrically charged molecules that are affected by external electric fields. Thus, when an electric field is applied, it creates interruption of metabolism, reduction of enzyme activity, weakening surface attachment, and nonuniform electrolyte distribution, resulting in separation from the surface. Thus, integration of the BE on a toothbrush enables enhanced biofilm reduction. The toothbrush tested with mild gingival patient (N=40) at the department of dentistry, Ulsan University Hospital, South Korea. The patient used the BE toothbrush and non-BE toothbrush consecutively for 2 weeks in each, following their gingival index (GI) and plaque index (PI) measurement for the quantitative analysis. The results demonstrate 1.75 times more decrease of GI in BE toothbrush than the data of the non-BE device (p0.05). In conclusion, a new electromagnetic wave toothbrush has been successfully demonstrated especially in reduction of inflammation. Based on this work, we are going to further develop a medical device that dedicated on the treatment of inflammation.

We report the complete genome sequence of a novel nucleorhabdovirus, apple rootstock virus A (ApRVA), isolated from Malus spp. in South Korea. ApRVA has a 14,043-nt single-stranded negative-sense RNA genome. In the antigenome sense, it contains seven open reading frames, encoding the putative nucleocapsid protein, phosphoprotein, cell-to-cell movement protein, matrix protein, glycoprotein, RNA-dependent RNA polymerase, and an additional hypothetical protein, the gene for which is located between the genes for the matrix protein and glycoprotein. The complete genome sequence of ApRVA showed 47.45% nucleotide sequence identity to that of black currant-associated rhabdovirus 1. The genome organization, phylogenetic relationships, and sequence similarities to other nucleorhabdoviruses suggest that ApRVA is a new member of the genus Nucleorhabdovirus.

Complete genome sequences of two cycas necrotic stunt virus (CNSV) isolates from Paeonia suffruticosa and Daphne odora were determined. Phylogenetic trees and pairwise comparisons using complete RNA1- and RNA2-encoded polyproteins showed that the two CNSV isolates are divergent (83.19%-89.42% in polyprotein 1 and 73.61%-85.78% in polyprotein 2). A comparative analysis based on taxonomic criteria for the species demarcation of nepoviruses confirmed that they are not new species but distinct variants. This is the first report of the complete genome sequences of CNSV detected in P. suffruticosa and D. odora, and the first report of CNSV infecting P. suffruticosa.

A new member of the genus Closterovirus was detected in Platycodon grandiflorus using high-throughput RNA sequencing analysis. The complete genome sequence of this new virus isolate, tentatively named "platycodon closterovirus 1" (PlaCV1), comprises 16,771 nucleotides with nine predicted open reading frames (ORFs) having the typical closterovirus genome organization. PlaCV1 shares 37%-50% nucleotide sequence identity with other known closterovirus genome sequences. The putative RNA-dependent RNA polymerase (RdRp), heat shock protein 70-like protein (HSP70h), viral heat shock protein 90-like protein (HSP90h), minor coat protein (CPm), and coat protein (CP) show 47-68%, 39-66%, 24-52%, 21-57%, and 16-35% amino acid sequence identity, respectively, to homologous proteins in previously identified closteroviruses, suggesting that it represents a distinct, new species in the genus. Phylogenetic analysis of HSP70h sequences places PlaCV1 alongside other members of the genus Closterovirus in the family Closteroviridae. To our knowledge, this study is the first report of the complete genome sequence of PlaCV1 infecting P. grandiflorus in the Republic of Korea.

Wheat streak mosaic virus (WSMV), a member of the genus Tritimovirus in the family Potyviridae, has been designated as a plant quarantine pathogen in South Korea. Several diagnostic methods can be applied to diagnose viral infections in plants, but polymerase chain reaction and enzyme-linked immunosorbent assay, which can identify viral species with speed and accuracy, are mainly used in Korean plant quarantine. Many variants of different viral species with highly divergent genomic sequences are constantly being discovered by high-throughput sequencing technology. This means that previously established primers may no longer be suitable for diagnostic use. In this study, we developed a reverse transcription polymerase chain reaction assay for detecting WSMV isolates/strains using all of the WSMV sequences available in NCBI GenBank. All 13 primer sets were able to produce amplicons of the expected sizes from WSMV-infected samples. To check whether nonspecific reactions occur, some closely related viruses (one tritimovirus and five potyviruses) and target imported plants (wheat, maize, oat, and proso millet) were tested. Consequently, four primer sets, which did not produce nonspecific bands, were finally selected among the 13 primer sets. Concentration-dependent amplification tests showed that the four primer sets are adequate for use in the diagnosis of WSMV in Korean plant quarantine. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13337-020-00646-3) contains supplementary material, which is available to authorized users.

The complete genome sequence of a new polerovirus found naturally infecting Artemisia princeps, artemisia virus B (ArtVB), was determined using high-throughput sequencing. The ArtVB genome comprises 6,141 nucleotides and contains six putative open reading frames (ORF0 to ORF5) with a genome structure typical of poleroviruses. A multiple sequence alignment showed that the complete ArtVB genome shares 50.98% nucleotide sequence identity with ixeridium yellow mottle virus 1 (IxYMaV-1, GenBank accession no. KT868949). ArtVB shares the highest amino acid sequence identity in P0 and P3-P5 (21.54%-51.69%) with other known poleroviruses. Phylogenetic analysis indicated that ArtVB should be considered a member of a new species within the genus Polerovirus, family Luteoviridae.

The complete genomic sequence of achyranthes virus A (AcVA), from an Achyranthes bidentata Blume plant in South Korea, was determined. The genomic RNA has 9491 nucleotides (nt), excluding the 3'-terminal poly(A) tail and contains an open reading frame typical of members of the genus Potyvirus, family Potyviridae, encoding a large putative polyprotein of 3103 amino acids (aa). Pairwise comparisons showed that the AcVA sequence shares 47.81-57.78% nt sequence identity at the complete genome level, 41.89-56.41% aa sequence identity at the polyprotein level, and 50-63.8% aa sequence identity at the coat protein level with other members of genus Potyvirus. These pairwise comparison values are below the species demarcation cutoff for the family Potyviridae. Our results therefore suggest that this virus should be regarded as a novel member of the genus Potyvirus, tentatively named "achyranthes virus A".

In 2010, foliar symptoms were observed in the paprika leaves in Jinju city, Korea. Beet western yellows virus (BWYV) was identified in paprika by using the large-scale oligonucleotide chip assay. To investigate the occurrence of BWYV, a survey was performed on various crops, including paprika, from 2011 to 2014. Further, the presence of BWYV was consistently verified through literature survey from 2015 to 2017. BWYV infection has been identified in Solanaceae crops (bell pepper, hot pepper, and paprika), various weeds, and green peach aphids and it occurs on a nationwide scale. Cultivation using organic methods involved natural enemies and showed a high BWYV infection rate, which was more than that for conventional cultivation methods in greenhouse. The complete genome sequence of BWYV isolated from paprika was determined for the first time. The genome of the BWYV-Korea isolate consists of 5750 nucleotides and has six open reading frames. Sequence identity results showed maximum similarity between the BWYV-Korea isolate and the BWYV LS isolate (identity ＞ 90%). This study is the first report of BWYV infecting paprika in Korea. The survey revealed that BWYV is naturalized in the domestic ecology of Korea.

The complete nucleotide sequence of peach virus D (PeVD) from Prunus persica was determined. The PeVD genome consists of 6,612 nucleotides excluding the 3' poly(A) tail and contains a single open reading frame coding for a polyprotein of 227 kDa. Sequence comparisons and phylogenetic analysis revealed that PeVD is most closely related to viruses in the genus Marafivirus, family Tymoviridae. The complete nucleotide and CP amino acid sequences of PeVD were most similar (51.1-57.8% and 32.2-48.0%, respectively) to members of the genus Marafivirus, suggesting that PeVD is a new member of this genus.

In July 2013, soybean (Glycine max) plants at the research field in Daegu, South Korea, showed virus-like symptoms, such as mosaic, mottle, yellowing, and stunting. Overall, there were approximately 1% of soybean plants that showed these symptoms. Sixteen soybean samples were collected based on visual symptoms and subjected to laboratory characterization. Total RNA was extracted from each sample with the Tri Reagent (Molecular Research Center, Cincinnati, OH) and cDNA was synthesized using random N25 primer with RevertAid Reverse Transcriptase (Thermo Scientific, Waltham, MA), according to the manufacturers' instructions. All samples were tested by PCR with Prime Taq Premix (2X) (GeNet Bio, Daejeon, Korea) and primer sets specific to Soybean mosaic virus (SMV; 5′-CATATCAGTTTGTTGGGCA-3′ and 5′-TGCCTATACCCTCAACAT-3′), Peanut stunt virus (PSV; 5′-TGACCGCGTGCCAGTAGGAT-3′ and 5′-AGGTDGCTTTCTWTTGRATTTA-3′), Soybean yellow mottle mosaic virus (SYMMV; 5′-CAACCCTCAGCCACATTCAACTAT-3′ and 5′-TCTAACCACCCCACCCGAAGGATT-3′), and Soybean yellow common mosaic virus (SYCMV; 5′-TTGGCTGAGAGGAGTGGCTT-3′ and 5′-TGCGGTCGTGTAGTCAGTG-3′). Among 16 samples tested, five were positive for SMV and two for SYMMV. Three samples were found infected by both SMV and SYMMV and four by both SMV and PSV. Since two of the symptomatic samples were not infected by viruses described above, a pair of primers specific to Peanut mottle virus (PeMoV; 5′-GCTGTGAATTGTTGTTGAGAA-3′ and 5′-ACAATGATGAAGTTCGTTAC-3′) was tested (1). All 16 samples were subjected to RT-PCR with primers specific to PeMoV. Four were found positive for PeMoV. Two of them were already found infected with SYMMV. In order to identify the complete nucleotide sequences of PeMoV coat protein (CP), another primer set (5′-TGAGCAGGAAAGAATTGTTTC-3′ and 5′-GGAAGCGATATACACACCAAC-3′) was used. RT-PCR product was cloned into RBC TA Cloning Vector (RBC Bioscience, Taipei, Taiwan) and the nucleotide sequence of the insert was determined by Macrogen (Seoul, Korea). CP gene of the PeMoV (GenBank Accession No. KJ664838) showed the highest nucleotide sequence identity with PeMoV isolate Habin (KF977830; 99% identity), and the highest amino acid identity with GenBank Accession No. ABI97347 (100% identity). In order to fulfill Koch's postulates, several G. max cv. Williams 82 were inoculated with the extracts of PeMoV-infected leaf tissue. At 14 days post-inoculation, plants showed systemic mottle symptoms. These symptomatic plants were subjected to RT-PCR, and the nucleotide sequences of the PCR product were found identical to that of the virus in the inoculum. To our knowledge, this is the first report of soybean-infecting PeMoV, a member of the genus Potyvirus in the family Potyviridae, in South Korea. Reference: (1) R. G. Dietzgen et al. Plant Dis. 85:989, 2001.

Deep sequencing has generated 52 contigs derived from five viruses; Apple chlorotic leaf spot virus (ACLSV), Apple stem grooving virus (ASGV), Apple stem pitting virus (ASPV), Apple green crinkle associated virus (AGCaV), and Apricot latent virus (ApLV) were identified from eight apple samples showing small leaves and/or growth retardation. Nucleotide (nt) sequence identity of the assembled contigs was from 68% to 99% compared to the reference sequences of the five respective viral genomes. Sequences of ASPV and ASGV were the most abundantly represented by the 52 contigs assembled. The presence of the five viruses in the samples was confirmed by RT-PCR using specific primers based on the sequences of each assembled contig. All five viruses were detected in three of the samples, whereas all samples had mixed infections with at least two viruses. The most frequently detected virus was ASPV, followed by ASGV, ApLV, ACLSV, and AGCaV which were withal found in mixed infections in the tested samples. AGCaV was identified in assembled contigs ID 1012480 and 93549, which showed 82% and 78% nt sequence identity with ORF1 of AGCaV isolate Aurora-1. ApLV was identified in three assembled contigs, ID 65587, 1802365, and 116777, which showed 77%, 78%, and 76% nt sequence identity respectively with ORF1 of ApLV isolate LA2. Deep sequencing assay was shown to be a valuable and powerful tool for detection and identification of known and unknown virome in infected apple trees, here identifying ApLV and AGCaV in commercial orchards in Korea for the first time.

Four samples of Plantago asiatica showing mottle or mottled yellowing symptoms were collected and three of them were confirmed to be infected with Plantago asiatica mosaic virus (PlAMV) by Illumina HiSeq 2500 paired-end RNA sequencing. Consequently, the complete genomic sequence of a Korean isolate of PlAMV (PlAMV isolate Gunwi) from P. asiatica was determined. The nucleotide sequence of PlAMV-Gunwi was most closely related to a Russian isolate of PlAMV (PlAMV-Ru) collected from P. asiatica but highly dissimilar (~23 %) to other isolates of PlAMV from other plant species. Pairwise comparisons revealed that the complete replicase protein and coat protein of PlAMV-Gunwi share 85.15–91.20 and 84.54–90.82 % amino acid sequence identities, respectively, to the corresponding proteins of other PlAMV isolates. To our knowledge, this is the first report of the natural infection of P. asiatica by PlAMV in Korea, and the divergent PlAMV isolate Gunwi expands our understanding of the epidemiology of PlAMV.

The complete nucleotide sequence of a new barley polerovirus, tentatively named barley virus G (BVG), which was isolated in Gimje, South Korea, has been determined using an RNA sequencing technique combined with polymerase chain reaction methods. The viral genomic RNA of BVG is 5,620 nucleotides long and contains six typical open reading frames commonly observed in other poleroviruses. Sequence comparisons revealed that BVG is most closely related to maize yellow dwarf virus-RMV, with the highest amino acid identities being less than 90 % for all of the corresponding proteins. These results suggested that BVG is a member of a new species in the genus Polerovirus.

Apple (Malus domestica) is economically important and widely grown in South Korea. Many viruses and viroids infect apple trees, reducing the productivity of orchards (Baek et al. 2019; James et al. 2013). Apple stem grooving virus (ASGV), apple mosaic virus (ApMV), apple latent spherical virus (ALSV), apple chlorotic leaf spot virus (ACLSV), apple green crinkle associated virus (AGCaV), apple stem pitting virus (ASPV), and apple scar skin viroid (ASSVd) are the major viral pathogens that infect apple (Cho et al. 2016). Malus domestica virus A (MdoVA), a putative member of Velarivirus (Closteroviridae), has been identified from an apple tree in the Czech Republic (Koloniuk et al. 2019; Rubio et al. 2013). From August to September 2018, 11 asymptomatic and 11 symptomatic apple leaf samples showing mild mosaic symptoms, necrotic leaf spots, mottling, leaf distortion, or yellowing were collected from Uiseong (n = 4; cv. Fuji), Yeongcheon (n = 4; cv. Hongro), and Jangsu (n = 14; Fuji = 8 and Hongro = 6). Total RNA was extracted from two symptomatic and two asymptomatic samples using a HiYield Total RNA Mini Kit (Plant; RBC Bioscience). Four cDNA libraries were constructed and sequenced by the Illumina HiSeq4000 system at Macrogen. A total of 219,160,492 paired-end raw reads were generated from the four libraries with the Illumina HiSeq4000 sequencer. The raw reads were quality-trimmed and assembled into 387,422 contigs of 201 to 16,053 nt using Trinity v2.1.1 software. BLASTn analysis of the contigs indicated that the selected samples were infected with several apple viruses, including ASPV, ASGV, ACLSV, AGCaV, and apple hammerhead viroid (AHVd). A BLASTn search of the obtained contigs also detected seven contigs (741 to 5,138 nt) related to MdoVA (GenBank MN548734). The nucleotide (nt) similarities between these contigs and the MdoVA reference sequence were 88.1 to 93.9%. Specific primer sets (i.e., MdoVA-SK_F3 [nt position 2,496 to 2,515]: 5′-GTTGGCTCATGACAGTTTGC-3′; MdoVA-SK_R3 [3,722 to 3,741]: 5′-GGATGTGGTAAGGTAGTACA-3′; MdoVA-SK_F9 [9,269 to 9,288]: 5′-GTACATGAGGACTATCCTCG-3′; and MdoVA-SK_R9 [10,464 to 10,483]: TCTGACTTTCCTGAGGAACC-3′) were designed to confirm the presence of MdoVA in some of the apple leaf samples. Total RNA was extracted from the 22 samples individually and subjected to RT-PCR using SuPrimeScript RT-PCR Premix (GeNet Bio) with MdoVA-specific primers. The expected fragments of 1,246 and 1,215 bp were only detected in the sole asymptomatic Fuji sample that was included for high-throughput sequencing, suggesting that the virus was not associated with any of the symptoms observed. This sample was used to determine the complete genome sequence of this South Korea MdoVA isolate (MdoVA-SK). Specific primer sets were designed based on the seven contig sequences. All designed primer pairs successfully produced overlapping RT-PCR fragments of the expected size from the sample. PCR products were purified with an AccuPrep PCR Purification Kit (BIONEER) and cloned into an RBC TA cloning vector (RBC Bioscience). At least four clones of each fragment were Sanger sequenced. The obtained overlapping sequences were consistent with the original assembled contigs from paired-end RNA sequencing. The 5′- and 3′-terminal sequences were obtained using a 5′- and 3′-Rapid Amplification of cDNA Ends kit (RACE version 2.0; Invitrogen) based on the MdoVA-SK specific 5′ and 3′ RACE primers. The complete genome sequence of MdoVA-SK (17,004 nt) was assembled using DNAMAN v5.2.10 (Lynnon Biosoft) and deposited in GenBank (MN901923). BLASTn analysis showed that MdoVA-SK shared 91.3% sequence identity at the complete genome level with the sole MdoVA isolate (J1, MN548734) in GenBank. At the individual gene level, the identities between the isolates were 88.6 to 99.0%. This is the first report of MdoVA naturally infecting apple trees in South Korea and the second report in the world. Although the virus did not appear to be associated with any disease symptoms in apple trees, more research is needed to understand its biological and pathological properties.

High-throughput sequencing revealed a tentative new badnavirus infecting Codonopsis lanceolata, provisionally named Codonopsis vein clearing virus (CoVCV). The complete 8,112-nt CoVCV genomic DNA sequence (GenBank accession: MK044821) comprises three open reading frames (ORFs) encoding conserved domains, with typical features of badnaviruses. Additionally, BLASTn searches indicated the CoVCV genome sequence is most similar to the grapevine vein clearing virus (GVCV) genome (72% identity and 46% query coverage). Moreover, the polyprotein encoded in CoVCV ORF3 is most similar to the corresponding protein of GVCV, with 60% amino acid sequence identity (89% query coverage). These results suggest that CoVCV is a new member of the genus Badnavirus in the family Caulimoviridae.

The complete genomic RNA sequence of a tentative new umbravirus from Patrinia scabiosaefolia, tentatively named "patrinia mild mottle virus" (PatMMoV), was determined. The genome of PatMMoV consists of 4,214 nucleotides and has a typical umbravirus genome organization with four open reading frames. BLAST searches showed that the complete nucleotide sequence of PatMMoV had the highest identity (72%; 50% query coverage) to Ixeridium yellow mottle-associated virus 2 (IxYMaV-2; an unclassified umbravirus). In addition, phylogenetic analysis and pairwise comparisons showed that PatMMoV and IxYMaV-2 were the most closely related and placed in the same clade within a group of umbraviruses. These results suggest that PatMMoV is a putative new member of the genus Umbravirus in the family Tombusviridae.

The complete 9,556-nt genomic sequence of a putative new potyvirus, tentatively named “platycodon mild mottle virus” (PlaMMV), infecting Platycodon grandiflorum was determined (GenBank accession no. MH779625). A single large open reading frame (ORF; nt 121-9360, 3079 aa) encoding a polyprotein that was predicted to be cleaved into 10 mature proteins was detected. By comparison with other potyviruses, nine cleavage sites and conserved domains/motifs were identified. PlaMMV also has a highly conserved GA6 motif (nt 2857-2863) that could enable a small overlapping ORF (PIPO) embedded within the P3 cistron to be translated by a viral polymerase slippage mechanism. A BLAST analysis showed that the complete PlaMMV nucleotide sequence shared 70% identity (40% query coverage) with carrot thin leaf virus (JX156434) and that the complete amino acid sequence of the PlaMMV polyprotein shared 54% identity (98% query coverage) with Thunberg fritillary mosaic virus (CAI59123). These results suggest that PlaMMV is a new member of the genus Potyvirus in the family Potyviridae.

Maize yellow mosaic virus (MaYMV) is a novel putative polerovirus recently found in maize (Zea mays) in China and has been detected not only in Asia but also in Africa and South America (Chen et al. 2016; Goncalves et al. 2017; Palanga et al. 2017). In addition, MaYMV infection has also been reported in sugarcane (Saccharum spp.) and itch grass (Rottboellia cochinchinensis) (Yahaya et al. 2017). Recently, the first occurrence of MaYMV in South Korea has been reported from Panicum miliaceum and Sorghum bicolor using high-throughput RNA sequencing (Lim et al. 2018). Therefore, to confirm whether MaYMV exists in maize in South Korea, a total of 26 maize samples with yellowing, yellow stripe, and chlorosis symptoms were collected from three provinces (Chungcheongbuk-do, Gangwon-do, and Gyeongsangnam-do) in August and September 2017. Total RNA was extracted from all samples using the easy-spin Total RNA Extraction Kit (iNtRON Biotechnology, Seongnam, Korea). Reverse transcription polymerase chain reaction (RT-PCR) was conducted using SuPrimeScript RT-PCR Premix (GeNet Bio, Daejeon, Korea) and MaYMV-specific primers (Chen et al. 2016), producing an amplicon with expected size of 753 bp, using protocols described previously (Lim et al. 2018). Five out of six samples collected from Chungcheongbuk-do, five out of 10 samples collected from Gangwon-do, and one out of 10 samples collected from Gyeongsangnam-do were confirmed to be infected with MaYMV. One RT-PCR fragment of MaYMV from each region (Gangwon-do, Chungcheongbuk-do, and Gyeongsangnam-do) was directly sequenced (Macrogen, Daejeon, Korea), and the sequences were deposited in GenBank under accession numbers MG020517, MG020518, and MG020519, respectively. The partial nucleotide sequences of MaYMV determined in each sample collected from Chungcheongbuk-do and Gyeongsangnam-do were identical, and these nucleotide sequences differed by one nucleotide from that identified in the sample collected from Gangwon-do. The NCBI BLAST search showed that the three MaYMV isolates shared high nucleotide sequence homology (99 to 100%) with isolates Yunnan3 (KU291099) and Yunnan4 (KU291100) from maize in China. The MaYMV-infected samples showed yellow stripe and chlorosis symptoms, which tend to first appear on the tip and edge of the leaves and become worse. However, further studies are needed, because it is uncertain whether the symptoms were caused by a single infection with MaYMV. To the best of our knowledge, this is the first report of MaYMV on Z. mays in South Korea.

A total of nine contigs related to caulimovirus-like sequences were detected using high-throughput paired-end RNA sequencing. An attempt to find the plant sample infected with this type of virus identified the medicinal plant Atractylodes macrocephala Koidzumi showing mild mottle symptoms. Subsequently, the complete DNA genome sequence of the Atractylodes virus was determined. The 8,105-nt genome of the virus was composed of six open reading frames and displayed the highest nucleotide sequence identity (70%) with soybean Putnam virus. Based upon the symptoms observed on the source plant, we propose to refer to this new member of the genus Caulimovirus as atractylodes mild mottle virus.

We determined the complete genome sequences of two isolates of cnidium vein yellowing virus (CnVYV-1 and -2) that co-infected all field samples collected from Cnidium officinale in Korea. Unlike CnVYV-2, however, CnVYV-1 was sap-transmissible to Nicotiana benthamiana. CnVYV-1 and -2 have bipartite genomes of 7,263 and 3,110 nucleotides and 7,278 and 3,112 nucleotides, respectively, excluding the poly(A) tails. Phylogenetic analysis of the CnVYV-1 and -2 sequences indicated close relationships to strawberry latent ringspot virus, an unassigned member of the family Secoviridae. CnVYV-1 and CnVYV-2 are closely related viruses that may represent a tentative new species of the family Secoviridae.

The complete genome sequence of a member of a distinct species of the genus Carlavirus in the family Betaflexiviridae, tentatively named atractylodes mottle virus (AtrMoV), has been determined. Analysis of its genomic organization indicates that it has a single-stranded, positive-sense genomic RNA of 8866 nucleotides, excluding the poly(A) tail, and consists of six open reading frames typical of members of the genus Carlavirus. The individual open reading frames of AtrMoV show moderately low sequence similarity to those of other carlaviruses at the nucleotide and amino acid sequence levels. Pairwise comparison and phylogenetic analysis suggest that AtrMoV is most closely related to chrysanthemum virus B.

The complete genomic sequence of lychnis mottle virus (LycMoV) from a Lychnis cognata plant was determined. LycMoV has a bipartite genome consisting of RNA1 (7,428 nt) and RNA2 (3,734 nt). Species in the family Secoviridae are demarcated based on their amino acid similarities in the protease-polymerase and coat protein. In LycMoV, these proteins share 90 % and 63 % sequence similarity, respectively, with the most closely related virus, strawberry latent ringspot virus, which is a member of the family Secoviridae but has not been assigned to a genus. Therefore, LycMoV is a tentative new virus of the family Secoviridae.

Garlic generally becomes coinfected with several types of viruses belonging to the Potyvirus, Carlavirus, and Allexivirus genera. These viruses produce characteristically similar symptoms, they cannot be easily identified by electron microscopy (EM) or immunological detection methods, and they are currently widespread around the world, thereby affecting crop yields and crop quality adversely. For the early and reliable detection of garlic viruses, virus-specific sets of primers, including species-specific and genus-specific primers were designed. To effectively detect the twelve different types of garlic viruses, primer mixtures were tested and divided into two independent sets for multiplex polymerase chain reaction (PCR). The multiplex PCR assays were able to detect specific targets up to the similar dilution series with monoplex reverse transcription (RT)-PCR. Seventy-two field samples collected by the Gyeongbuk Agricultural Technology Administration were analyzed by multiplex RT-PCR. All seventy two samples were infected with at least one virus, and the coinfection rate was 78%. We conclude that the simultaneous detection system developed in this study can effectively detect and differentiate mixed viral infections in garlic.

Soybean yellow common mosaic virus (SYCMV) was recently reported from Korea, and a subsequent survey of soybean fields found that SYCMV, Soybean yellow mottle mosaic virus (SYMMV), and Soybean mosaic virus (SMV) infections were widespread. SYCMV has recently been developed into a Virus Inducing Gene Silencing (VIGS) vector for use as a reverse genetics tool for soybean, and here we report a modified SYCMV VIGS vector containing a new restriction enzyme site in the 3' non-coding region into which we inserted the Gateway system. Ultrasonically generated c.300 bp random fragments of Glycine max cDNA were inserted into the SYCMV VIGS vector, and individual colonies containing G. max cDNA were inocu- lated to cultivar Williams 82. We monitored the phenotype of inoculated soybean, and selected obvious visi- ble phenotypes caused by SYCMV-induced gene silencing which could enable annotation of gene function of unknown gene fragments inserted into SYCMV. Here, we describe development of a high-throughput SYCMV VIGS vector for gene function identification in soybean.

The complete genomic sequence of a novel putative member of the genus Potyvirus was detected from Callistephus chinensis (China aster) in South Korea. The genomic RNA consists of 9,859 nucleotides (excluding the 3' poly(A) tail) and contains the typical open reading frame of potyviruses, encoding a putative large polyprotein of 3,154 amino acids. The Callistephus virus is most closely related to plum pox virus and members of the ApVY subgroup which showed 50-52 % polyprotein amino acid sequence identity. These results suggest that the Callistephus virus is a novel member of the genus Potyvirus, tentatively named "callistephus mottle virus" (CalMV).

The complete genome sequence of a Papaya ringspot virus (PRSV) isolate from South Korea (SK) infecting squash ( Cucurbita pepo ) was obtained using paired-end RNA sequencing. A BLASTn search of the PRSV SK isolate full-genome sequence showed nucleotide sequence identity ranging from 81% to 83% with previously reported PRSV isolates (GenBank accession numbers KX655874 and EF017707).

The complete genome sequence of an Uiseong isolate of barley virus G (BVG) on proso millet plants in a field in South Korea was determined by RNA sequencing and Sanger sequencing. To our knowledge, this is the first complete genome sequence report of BVG infecting proso millet in South Korea.