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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.

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".

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.

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.

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.

Throughout year 2015 to 2016, 101 proso millet and 200 sorghum samples were collected from five provinces in South Korea. The samples were subjected to paired-end RNA sequencing and further analyzed by RT-PCR. The results indicated that Rice black-streaked dwarf virus (RBSDV) was detected from sorghum collected in Gyeongsang province. The other four viruses, including RBSDV, Rice stripe virus (RSV), Barley virus G (BVG), and Cereal yellow dwarf virus (CYDV), were detected from proso millet. Among four viruses, both RSV and RBSDV were identified high frequency from proso millet collected from Gyeongsang province. Otherwise, BVG was nearly equally identified from five provinces, suggesting that the virus was supposedly widespread nationwide. RBSDV was first identified from both proso millet and sorghum in South Korea. The other virus annotated CYDV identified proso millet was shown to have relatively low identities compared to CYDV previously reported, suggesting that the virus might be new member of Polerovirus.

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.

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.

Two novel viruses, isolated in Bonghwa, Republic of Korea, from an Ixeridium dentatum plant with yellowing mottle symptoms, have been provisionally named Ixeridium yellow mottle-associated virus 1 (IxYMaV-1) and Ixeridium yellow mottle-associated virus 2 (IxYMaV-2). IxYMaV-1 has a genome of 6,017 nucleotides sharing a 56.4% sequence identity with that of cucurbit aphid-borne yellows virus (genus Polerovirus). The IxYMaV-2 genome of 4,196 nucleotides has a sequence identity of less than 48.3% with e other species classified within the genus Umbravirus. Genome properties and phylogenetic analysis suggested that IxYMaV-1 and -2 are representative isolates of new species classifiable within the genus Polerovirus and Umbravirus, respectively.

In May 2013, an angel’s trumpet leaves showing mosaic and malformation symptoms were collected from Suwon city, Gyeonggi-do. An analysis of the collected sample by transmission electron microscopy observation showed filamentous rod particles of 720-800 nm in length. On the basis of the these observations, we performed PCR against three reported Potyviruses (Brugmansia mosaic virus, Colombian datura virus and Brugmansia suaveolens mottle virus), and the sample was positive for BruMV. Pathogenicity and host range test of BruMV was determined by mechanical inoculation. Solanaceae (tobacco, tomato and eggplant) and Amaranthaceae (ground cherry) appeared typical virus symptoms. To determine coat protein of this virus, we designed specific primer pairs, and performed PCR amplification, cloning, and sequencing. Phylogenetic analysis showed that BruMV-SW was most closely related to BruMV isolate SK. Comparison of the BruMV-SW coat protein nucleotide sequences showed 92% to 99% identities to the other BruMV isolates.

RNA from a Chinese cabbage plant (Brassica campestris ssp. pekinensis) showing leaf malformation and mottling was labeled and hybridized to a DNA chip capable of detecting plant viruses and viroids. Probes specific for beet mild yellowing virus (BMYV) and beet western yellows virus (BWYV) yielded positive results, suggesting that the plant was infected by a polerovirus. Primers designed from the sequences of the positive probes were used to amplify and sequence one portion of the viral genome. This sequence showed a 90 % or greater identity to several poleroviruses, including BMYV, BWYV, beet chlorosis virus (BChV) and turnip yellows virus (TuYV). The complete genome sequence of the Chinese cabbage-infecting polerovirus consisted of 5,666 nt and was most closely related to brassica yellows virus (BrYV; 94 % identity). The virus was named BrYV-Cheongsong (BrYV-CS). However, ORF3, ORF4 and the 5' half of ORF5 of BrYV-CS were more closely related to those of TuYV, BWYV, BChV and BMYV than to those of BrYV. Interestingly, a recombination event (positions 3531-4819 in BrYV-CS) was detected when this sequence was aligned with those of BrYV and TuYV. This region showed the highest sequence identity to that of TuYV (94 % identity) and had greater than 93 % identity to those of BWYV, BChV and BMYV, but it shared only 81 % identity with that of BrYV. Taken together, the genomes of BrYV-CS and BrYV are closely related. However, the structural genes in the 3' half of the genome of BrYV-CS are more closely related to those of other poleroviruses.

In August and September 2018, apple trees of ‘Fuji’ and ‘Hongro’ cultivars looked abnormal with poor growth. Suspecting infection by viruses, leaf samples were collected from a total of 22 Fuji and Hongro apple trees in Uiseong, Yeongcheon, and Jangsu in Korea. To perform comparative analysis of virus infections, leaves from a healthy and a diseased tree of Fuji and Hongro apple cultivars were chosen. Total RNA was extracted from each of the four selected samples with a HiYield Total RNA Mini Kit (Plant; RBC Bioscience, Taipei, Taiwan), and paired-end RNA sequencing was conducted using an Illumina HiSeq4000 system at Macrogen (Daejeon, Korea). Analysis of the de novo assembled contigs showed that the apple trees were infected with several viruses including apple green crinkle associated virus, apple stem grooving virus, apple stem pitting virus, and apricot latent virus. In addition, the samples were also infected with apple hammerhead viroid (AHVd), recently identified in apple trees (Szostek et al. 2018; Zhang et al. 2014). AHVd was found in three samples but not in the abnormal-looking Hongro sample. We tried to determine if there is a correlation between AHVd infection and poor growth of apple trees. To detect AHVd in 22 apple leaf samples, AHVd-specific primers (5′-TTAGCCTTCCTGATGAGTCC-3′ and 5′-TGTGTCTACTTAAAGACTCAC-3′) were designed using the contig sequences and 18 AHVd sequences available in GenBank. RNA extracted from the leaf samples was subjected to reverse transcription (RT) polymerase chain reaction (PCR) using SuPrimeScript RT-PCR Premix (GeNet Bio, Daejeon, Korea) with the specific primer set. RT-PCR tests confirmed that 17 out of the 22 samples were infected with AHVd: five out of six healthy-looking Fuji samples, five out of six abnormal-looking Fuji samples, four out of five healthy-looking Hongro samples, and three out of five abnormal-looking Hongro samples. Thus, there was no correlation between presence of AHVd and poor growth. To obtain AHVd sequences from healthy-looking and abnormal-looking samples of the two cultivars, Sanger sequencing of RT-PCR amplicons was conducted at Macrogen. Each RT-PCR amplicon was purified using an AccuPrep PCR Purification Kit (BIONEER, Daejeon, Korea) and cloned into the RBC TA 100% query coverage) to AHVd isolate Italy2 (GenBank: MH049330). To the best of our knowledge, this is the first report on the presence of AHVd infecting apple trees in Korea. It is still not known if AHVd can cause any disease on apple or other orchard crop plants, and future studies need to address this.

Recently, several tentative new viruses have been discovered from apple trees (Malus pumila) using high-throughput sequencing (Liu et al. 2018; Rott et al. 2018); therefore, we have also investigated the existence of viruses infecting apple tree rootstocks, including new or emerging viral species, in Korea. Leaf samples, exhibiting yellowing and mottled symptoms, from 10 rootstocks (P1, Ottawa 11, Ottawa 13, Ottawa 5M, MM102, MM115, M5, M7, M10, and JM7) at the Apple Research Institute located in Gunwi and 20 rootstocks (M9) at a farmer’s apple orchard located in Cheongju were collected in September 2017. All samples were mixed into one pool, and an Illumina HiSeq2500 system was used to produce 100 bp paired-end RNA sequencing reads (a total of approximately 30 Gbp) at Theragen Etex Bio Institute (Suwon, Korea). Raw data treatment was performed using SG-VIPdb at SeqGenesis (Daejeon, Korea), as described by Lim et al. (2015). The high-throughput RNA sequencing generated contigs derived from two recently discovered viruses (apple luteovirus 1 [ALV-1] and apple rubbery wood virus 1 [ARWV-1]) not reported in Korea, as well as five viruses (apple chlorotic leaf spot virus, apple green crinkle associated virus, apple stem grooving virus, apple stem pitting virus, and apricot latent virus [Cho et al. 2016]) already identified in apple trees cultivated in Korea. NCBI BLAST searches showed that the 6,361-nt ALV-1 contig (3,507 mapped reads) shared 94% nucleotide sequence identity (93% query coverage) with a PA8 isolate (GenBank accession no. MF120198), the only ALV-1 isolate publicly available, and the 1,062-nt ARWV-1 contig (576 mapped reads) shared 99% nucleotide sequence identity (99% query coverage) with a 4342-5 isolate (MF062138). To confirm the presence of ALV-1 and ARWV-1, ALV-1-specific primers, 5′-CAAATCGAGCAAGAGGAGGC-3′ and 5′-TTTGCCAGTCCAACAAGTCG-3′ (expected size of 443 bp), and ARWV-1-specific primers, 5′-CCACTTGCTTACTGTTTCTGC-3′ and 5′-TGGACCATGCAGCATATGAA-3′ (expected size of 617 bp), were designed using sequences of each contig. All individual leaf samples were used to extract total RNA with a WizPrep Plant RNA Mini Kit (wizbiosolutions, Seongnam, Korea), and reverse transcription polymerase chain reaction (RT-PCR) was conducted using SuPrimeScript RT-PCR Premix (GeNet Bio, Daejeon, Korea) with the specific primers. Of the 30 rootstock samples, six (M9) were found positive for ALV-1, and 11 (nine M9, one P1, and one JM7) were positive for ARWV-1. Among the virus-infected samples, four M9 samples were found to be coinfected by both ALV-1 and ARWV-1. One RT-PCR amplicon from a diagnosis of each viral infection was purified using an AccuPrep PCR Purification Kit (BIONEER, Daejeon, Korea) and sequenced at Macrogen (Daejeon, Korea). The NCBI BLAST searches of the amplicon sequences showed that the ALV-1 CJ15 isolate (from M9 rootstock; MH708164) had 97% nucleotide sequence identity to the PA8 isolate, and the ARWV-1 CJ11 isolate (from M9 rootstock; MH714536) had 98% nucleotide sequence identity to a 982-11 isolate segment S (MF062127). Because all ALV-1- and/or ARWV-1-infected samples were confirmed to be coinfected with more than two other viruses already identified in Korea mentioned above, a further survey is needed to clearly understand the symptoms caused by the viral infections. To the best of our knowledge, this is the first report on the existence of ALV-1 and ARWV-1 infecting apple tree rootstocks in Korea.

Soybean yellow mottle mosaic virus (SYMMV) and Soybean yellow common mosaic virus (SYCMV) were recently isolated in Korea, and it has not been reported how two viruses were dispersed in Korea. In 2012, we performed nationwide survey in subsistence soybean farming. Suspicious virus-infected infected leave were collected from the field and a total of 682 soybean tissue samples were assayed by RT-PCR using triplex primers detecting SYMMV, SYCMV, and Soybean mosaic virus (SMV). On hundred two samples showed SMV positive, and SYMMV and SYCMV were detected in 116 and 17 tissue samples, respectively. No sample showed double infection of SYMMV and SYCMV, but there were double infection tissues indicating two viruses positive of SMV plus SYMMV (5 tissue samples) and SMV plus SYCMV (1 tissue sample). Through this first subsistence soybean farming field survey, we assumed soybean viruses were originated from home seed production managed by farmer. Thus, in order to prevent possible seed transmission and further damage caused by virus transmission, virus-free commercial soybean seeds are recommended to be planted.

We determined the complete genome sequence of a South Korean (SK) isolate of Brugmansia mosaic virus (BruMV), which has recently been proposed to be a member of a new species in the genus Potyvirus. The genomic RNA of BruMV isolate SK is 9781 nucleotides in length (excluding the 3'-terminal poly (A)) and shares complete nucleotide and polyprotein amino acid sequence identities of 85.6 % and 93.1 %, respectively, with the type isolate (D-437) of BruMV described in the USA. To our knowledge, this is the first report providing evidence of considerable sequence variation in BruMV.

A virus isolate designated Angelica bushy stunt virus (AnBSV), provisionally representing a new species in the genus Caulimovirus, was discovered in the medicinal plant Angelica dahurica. The complete 8,300-nt genomic DNA of AnBSV had seven putative open reading frames containing conserved domains/motifs, which are typical features of caulimoviruses, and showed the greatest nucleotide sequence identity (74% identity and 27% query coverage) to a lamium leaf distortion virus isolate. Interestingly, the new caulimovirus exists as endogenous pararetroviral sequences in the host plant and is considered to have multiple defective plant genome-integrated copies that may lead to the generation of subgenomic DNA species.

Habenaria mosaic virus (HaMV), a member of the genus Potyvirus in the family Potyviridae , was first discovered from Habenaria radiata in Japan . The complete genomic sequence of a South Korean isolate (PA1) of HaMV infecting Plantago asiatica L. was determined with high-throughput RNA sequencing.

We report here the first complete genome sequence of Ornithogalum mosaic virus (OrMV) isolated from Taean, South Korea, in 2011, which was obtained by next-generation sequencing and Sanger sequencing. The sequence information provided here may serve as a potential reference for other OrMV isolates.

The complete genome sequence of ligustrum virus A (LVA) from a Ligustrum obtusifolium Sieb. & Zucc. plant was determined. The genomic RNA has 8,525 nucleotides, excluding the poly(A) tail, and consists of six open reading frames typical of members of the genus Carlavirus, family Betaflexiviridae. Phylogenetic analysis of the viral replicase and coat protein (CP) indicated that LVA is closely related to daphne virus S and helenium virus. The replicase and CP of LVA shared 44.73–52.35 % and 25.39–62.46 % amino acid identity, respectively, with those of other carlaviruses. These results suggest that LVA is a member of a distinct carlavirus species.

From Daphne odora Thunb., an ornamental shrub in the Republic of Korea, a potyvirus was identified that has an RNA genome of 9,448 nucleotides (excluding the 3′-terminal poly(A) tail) encoding a polyprotein of 3,065 amino acids, with nine putative protease cleavage sites producing ten proteins. Since this potyvirus shared the highest nucleotide sequence identity (91 %; query coverage 5 %) with the available partial sequence of daphne virus Y (DVY) from New Zealand (EU179854), it was considered a Korean isolate of DVY. This is the first molecular characterization of the complete genome sequence of a DVY isolate.

We report here the development of tobacco ringspot virus (TRSV)-based vectors for the transient expression of foreign genes and for the analysis of endogenous gene function in plants using virus-induced gene silencing. The jellyfish green fluorescent protein (GFP) gene was inserted between the TRSV movement protein (MP) and coat protein (CP) regions, resulting in high in-frame expression of the RNA2-encoded viral polyprotein. GFP was released from the polyprotein via an N-terminal homologous MP-CP cleavage site and a C-terminal foot-and-mouth disease virus (FMDV) 2 A catalytic peptide in Nicotiana benthamiana. The VIGS target gene was introduced in the sense and antisense orientations into a SnaBI site, which was created by mutating the sequence following the CP stop codon. VIGS of phytoene desaturase (PDS) in N. benthamiana, Arabidopsis ecotype Col-0, cucurbits and legumes led to obvious photo-bleaching phenotypes. A significant reduction in PDS mRNA levels in silenced plants was confirmed by semi-quantitative RT-PCR.

Maize yellow mosaic virus (MaYMV) is a tentative new Polerovirus, which was recently identified from maize (Zea mays) in China (Chen et al. 2016). MaYMV has also been reported to infect sugarcane (Saccharum spp.) and itch grass (Rottboellia cochinchinensis), and it has been reported in Asia, Africa and South America (Goncalves et al. 2017; Palanga et al. 2017; Yahaya et al. 2017). In this study, MaYMV was detected in Panicum miliaceum and Sorghum bicolor using Illumina HiSeq2500 system by Theragen Etex Bio Institute (Suwon, Korea) and SG-VIPdb by SeqGenesis (Daejeon, Korea), as described by Lim et al. (2015). Twenty samples of P. miliaceum and sixty three samples of S. bicolor were collected from July to September and from June to October, 2016, respectively, in South Korea. The high-throughput RNA sequencing of the samples mixed into one pool resulted in a single large contig (5606-nt) with nearly complete MaYMV genome coverage. The contig was assembled from a total of 234,537 reads; the maximum, minimum...

The complete genome sequence of a South Korean isolate of Rehmannia mosaic virus (ReMV) infecting Rehmannia glutinosa was determined through next-generation sequencing and Sanger sequencing. To our knowledge, this is the first report of a natural infection of R. glutinosa by ReMV in South Korea.

In 2014, we performed a nationwide survey in Korean radish fields to investigate the distribution and variability of Turnip mosaic virus (TuMV). Brassica rapa ssp. pekinensis sap-inoculated with three isolates of TuMV from infected radish tissue showed different symptom severities, whereas symptoms in Raphanus sativus were similar for each isolate. The helper component-protease (HC-Pro) genes of each isolate were sequenced, and phylogenetic analysis showed that the three Korean isolates were clustered into the basal-BR group. The HC-Pro proteins of these isolates were tested for their RNA silencing suppressor (VSR) activity and subcellular localization in Nicotiana benthamiana. A VSR assay by co-agroinfiltration of HC-Pro with soluble-modified GFP (smGFP) showed that HC-Pro of isolate R007 and R041 showed stronger VSR activity than R065. The HC-Pros showed 98.25 % amino acid identity, and weak VSR isolate (R065) has a single variant residue in the C-terminal domain associated with protease activity and self-interaction compared to isolates with strong VSR activity. Formation of large subcellular aggregates of GFP:HC-Pro fusion proteins in N. benthamiana was only observed for HC-Pro from isolates with strong VSR activity, suggesting that R065 ‘weak’ HC-Pro may have diminished self-association; substitution of the variant C-terminal residue largely reversed the HC-Pro aggregation and silencing suppressor characteristics. The lack of correlation between VSR efficiency and induction of systemic necrosis (SN) suggests that differences in viral accumulation due to HC-Pro are not responsible for SN.

Here, we report complete genome sequences of grapevine yellow speckle viroid 1 (GYSVd1) and hop stunt viroid (HSVd), members of the family Pospiviroidae , assembled from the transcriptome data generated from Ixeridium dentatum plants. To our knowledge, this is the first report of GYSVd1 and HSVd in I. dentatum .

A peach tree (Prunus persica) showing yellowing and mild mottle symptoms was analyzed using high-throughput RNA sequencing to determine the causal agent. A total of nine contigs similar to Little cherry virus 1 (LChV-1) were produced, and all the contigs showed nucleotide sequence identity (lower than 83 %) and query coverage (higher than 73 %) with LChV-1. The symptomatic peach sample was confirmed to be infected with LChV-1-like virus as a result of reverse transcription-polymerase chain reaction using primers designed based on sequences of the contigs. Occurrence of diseases caused by LChV-1 in Prunus species has been reported. Complete 16,931-nt genome of the peach virus composed of eight open reading frames was determined, and conserved domains including viral methyltransferase, viral helicase 1, RNA-dependent RNA polymerase (RdRp), heat shock protein 70 homologue (HSP70h), HSP90h and closterovirus coat protein (CP) were identified. Phylogenetic trees based on amino acid sequence alignments between the peach virus and members in the family Closteroviridae showed that the virus was most similar to LChV-1. Pairwise comparisons based on amino acid sequence alignments of three genes (RdRp, HSP70h and CP) between the peach virus and LChV-1 isolates showed the highest amino acid sequence identities, with 84.32 % for RdRp, 85.48 % for HSP70h and 80.45 % for CP. These results indicate that this is the first report for the presence of LChV-1 in South Korea and may be one of the first reports of natural infection of peach by LChV-1. Although it is not clear if LChV-1 YD isolate was responsible for specific symptoms observed, detection and characterization of the peach tree-infecting LChV-1 in South Korea would be useful in terms of the epidemiology of LChV-1.

The genome of tetterwort vein chlorosis virus (TVCV) from South Korea has been completely sequenced. Its genomic organization resembles those of other criniviruses, with several new features, indicating that TVCV is a member of a new species in the genus Crinivirus, family Closteroviridae. RNA1 contains 8467 nucleotides, with at least four opening reading frames (ORFs). ORF1a encodes a protein with predicted papain-like protease, methyltransferase, and helicase activities. ORF1b encodes a putative RNA-dependent RNA polymerase that is apparently expressed through a +1 ribosomal frameshift. RNA2 contains 8113 nucleotides encoding at least nine proteins, similar to most crinivirus RNA2s. The 3′ untranslated regions of the bipartite RNA genome share 82.1 % nucleotide sequence identity.

Tobacco ringspot virus (TRSV, genus Nepovirus), causes severe diseases in soybean and tobacco plants. TRSV-induced bud blight disease significantly reduced both the yield and quality of soybeans. The function of the encoded viral gene product involved in TRSV infection was unclear due to the limitation of reverse genetics studies on the viral genome. Here, we represent the successful construction of infectious full-length cDNA clones of TRSV genome (RNA1 and RNA2). The cDNAs of TRSV RNA1 and RNA2 were cloned into the binary vector pPZP211 immediately downstream of a double cauliflower mosaic virus 35S promoter and upstream of the nopaline synthase terminator. Seven days after agrobacterium-mediated co-inoculation of these two constructs, Nicotiana benthamiana plants developed a systemic infection with necrotic ringspot symptoms and weak stunting of the leaves, similar to that induced by natural TRSV. The systemic infection was confirmed by transmission electron microscopy and Western blot analysis. Simultaneously, soybean, tomato, and Arabidopsis ecotype Estland were mechanically inoculated with sap prepared from TRSV-agroinfiltrated N. benthamiana leaves, showing typical symptoms of bud blight, necrotic spots, and lethal systemic necrosis, respectively. The system developed herein will be an appealing way to determine TRSV viral gene functions and study host-TRSV interactions.

A new vector using Soybean yellow common mosaic virus (SYCMV) was constructed for gene function study or heterologous protein expression in soybeans. The in vitro transcript with a 5' cap analog m7GpppG from an SYCMV full-length infectious vector driven by a T7 promoter infected soybeans (pSYCMVT7-full). The symptoms observed in the soybeans infected with either the sap from SYCMV-infected leaves or pSYCMVT7-full were indistinguishable, suggesting that the vector exhibits equivalent biological activity as the virus itself. To utilize the vector further, a DNA-based vector driven by the Cauliflower mosaic virus (CaMV) 35S promoter was constructed. The complete sequence of the SYCMV genome was inserted into a binary vector flanked by a CaMV 35S promoter at the 5' terminus of the SYCMV genome and a cis-cleaving ribozyme sequence followed by a nopaline synthase terminator at the 3' terminus of the SYCMV genome (pSYCMV-full). The SYCMV-derived vector was tested for use as a virus-induced gene silencing (VIGS) vector for the functional analysis of soybean genes. VIGS constructs containing either a fragment of the Phytoene desaturase (PDS) gene (pSYCMV-PDS1) or a fragment of the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (RbcS) gene (pSYCMV-RbcS2) were constructed. Plants infiltrated with each vector using the Agrobacterium-mediated inoculation method exhibited distinct symptoms, such as photo-bleaching in plants infiltrated with pSYCMV-PDS1 and yellow or pale green coloring in plants infiltrated with pSYCMV-RbcS2. In addition, down-regulation of the transcripts of the two target genes was confirmed via northern blot analysis. Particle bombardment and direct plasmid DNA rubbing were also confirmed as alternative inoculation methods. To determine if the SYCMV vector can be used for the expression of heterologous proteins in soybean plants, the vector encoding amino acids 135-160 of VP1 of Foot-and-mouth disease virus (FMDV) serotype O1 Campos (O1C) was constructed (pSYCMV-FMDV). Plants infiltrated with pSYCMV-FMDV were only detected via western blotting using the O1C antibody. Based on these results, we propose that the SYCMV-derived vector can be used for gene function study or expression of useful heterologous proteins in soybeans.

A novel soybean-infecting sobemovirus termed Soybean yellow common mosaic virus (SYCMV) was characterized. The virus has a single, positive-strand RNA genome of 4152 nucleotides. The virus contains four putative open reading frames encoding P1 (78-566 nt), polyprotein ORF2a (524-2248 nt), polymerase domain ORF2b (1852-3417 nt), and CP (3227-4030 nt). The entire nucleotide sequence of SYCMV showed 31.2-71.3% nucleotide identity with the previously known eleven species of sobemovirus. In host range analysis of SYCMV, in which twenty one species and three different Nicotiana tabacum cultivars belonging to seven families were inoculated with the virus, SYCMV had a narrow host range, infecting only Glycine max and G. soja. Based on the obtained sequence, full-length clones of SYCMV were constructed. Symptoms produced by inoculation with clones were indistinguishable from those produced by inoculation with sap from symptomatic plants. Viral RNA accumulation of SYCMV was detected in the upper leaves by Northern blotting. This indicated that full-length clones of SYCMV were sufficient to produce disease symptoms. Genomic organization, the predicted amino acid sequence, and phylogenetic analyses with known sobemoviruses confirmed the assignment of SYCMV as a new member of the genus Sobemovirus.