RESUMO
BACKGROUND: Viruses have notable effects on agroecosystems, wherein they can adversely affect plant health and cause problems (e.g., increased biosecurity risks and economic losses). However, our knowledge of their diversity and interactions with specific host plants in ecosystems remains limited. To enhance our understanding of the roles that viruses play in agroecosystems, comprehensive analyses of the viromes of a wide range of plants are essential. High-throughput sequencing (HTS) techniques are useful for conducting impartial and unbiased investigations of plant viromes, ultimately forming a basis for generating further biological and ecological insights. This study was conducted to thoroughly characterize the viral community dynamics in individual plants. RESULTS: An HTS-based virome analysis in conjunction with proximity sampling and a tripartite network analysis were performed to investigate the viral diversity in chunkung (Cnidium officinale) plants. We identified 61 distinct chunkung plant-associated viruses (27 DNA and 34 RNA viruses) from 21 known genera and 6 unclassified genera in 14 known viral families. Notably, 12 persistent viruses (7 DNA and 5 RNA viruses) were exclusive to dwarfed chunkung plants. The detection of viruses from the families Partitiviridae, Picobirnaviridae, and Spinareoviridae only in the dwarfed plants suggested that they may contribute to the observed dwarfism. The co-infection of chunkung by multiple viruses is indicative of a dynamic and interactive viral ecosystem with significant sequence variability and evidence of recombination. CONCLUSIONS: We revealed the viral community involved in chunkung. Our findings suggest that chunkung serves as a significant reservoir for a variety of plant viruses. Moreover, the co-infection rate of individual plants was unexpectedly high. Future research will need to elucidate the mechanisms enabling several dozen viruses to co-exist in chunkung. Nevertheless, the important insights into the chunkung virome generated in this study may be relevant to developing effective plant viral disease management and control strategies.
Assuntos
Coinfecção , Nanismo , Vírus de Plantas , Vírus de RNA , Humanos , Viroma , Ecossistema , Cnidium/genética , RNA Viral/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Vírus de Plantas/genética , DNA , FilogeniaRESUMO
A putative new polerovirus, named "chrysanthemum virus D" (ChVD), was detected in a Chrysanthemum morifolium plant in South Korea. The virus was identified by high-throughput sequencing and confirmed by reverse transcription polymerase chain reaction. The entire ChVD genome is composed of 5,963 nucleotides and contains seven open reading frames (ORF0-5 and ORF3a), which are arranged similarly to those of other poleroviruses. These ORFs encode the putative proteins P0-5 and P3a, respectively. Pairwise amino acid sequence comparisons showed that the ChVD P0-5 and P3a proteins have 30.45-75% sequence identity to the corresponding proteins of other members of the genus Polerovirus. Since one of the species demarcation criteria for the genus Polerovirus is > 10% difference in the amino acid sequence of any gene product, the sequence comparisons indicate that ChVD represents a new species in this genus. Phylogenetic analysis of the P1-P2 and P3 amino acid sequences further indicate that ChVD is a novel polerovirus.
Assuntos
Chrysanthemum , Luteoviridae , Sequência de Bases , Filogenia , Chrysanthemum/genética , Genoma Viral , Doenças das Plantas , RNA Viral/genética , Luteoviridae/genética , Fases de Leitura Aberta , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
The family Convolvulaceae comprises approximately 50-60 genera with approximately 1600-1700 species, exhibiting a rich diversity of morphological characteristics and occupying a broad range of ecological habitats. High-throughput sequencing identified a tentative new virus in the family Geminiviridae infecting Calystegia sepium var. japonica in South Korea. The 2,706 nt long genome comprises six open reading frames (ORFs). The analysis of the nucleotide sequence of the genome and the putative amino acid sequences of ORFs indicated that the virus was closely related to the members of the family Geminiviridae. The genome organization of the virus was similar to that of members of the genus Topilevirus in terms of the number of ORFs and splicing signal. However, the virus, tentatively named bindweed mottle virus (BWMV), may not be assigned to current genera into the family Geminiviridae.
Assuntos
Geminiviridae , Genoma Viral , Fases de Leitura Aberta , Filogenia , Doenças das Plantas , Genoma Viral/genética , Geminiviridae/genética , Geminiviridae/classificação , Geminiviridae/isolamento & purificação , República da Coreia , Doenças das Plantas/virologia , Sequenciamento Completo do Genoma , Sequência de Bases , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
The complete nucleotide sequence of a newly discovered virus infecting Quercus aliena Blume, tentatively named "quercus leafroll virus" (QLRV), was determined through high-throughput and Sanger sequencing. The sequence comprises 3,940 nucleotides, has five open reading frames, and has a typical pelarspovirus genome organization, with neither 3' polyadenylation nor a 5' cap. The proteins encoded by QLRV share 17.9 to 44.2% amino acid sequence identity with known pelarspovirus proteins. The highest amino acid sequence identity values for the RNA-dependent RNA polymerase (RdRp) and coat protein were 67.5% and 55.2%, respectively, which are below the current thresholds for pelarspovirus species demarcation. On the basis of these results, we propose classifying QLRV as a new member of the genus Pelarspovirus, family Tombusviridae.
Assuntos
Quercus , Tombusviridae , República da Coreia , Sequência de Aminoácidos , NucleotídeosRESUMO
The complete nucleotide sequence of a novel gondre (Cirsium setidens)-infecting virus, provisionally named "cirsium virus A" (CiVA), was determined by high-throughput and Sanger sequencing, revealing a genome organization typical of fabaviruses. RNA1 and RNA2 are 5,828 and 3,478 nucleotides long, excluding the 3'-terminal poly(A) tails, each containing a single open reading frame. The highest sequence identity values for the CiVA coat protein and proteinase-polymerase, compared with known fabavirus sequences, were 59.09% and 69.68%, respectively, falling below the current thresholds for Fabavirus species demarcation. Our findings support classifying CiVA as a novel putative member of the genus Fabavirus, subfamily Comovirinae, family Secoviridae.
Assuntos
Cirsium , Fabavirus , Cirsium/genética , RNA Viral/genética , Genoma Viral , Proteínas do Capsídeo/genética , Filogenia , Fases de Leitura Aberta , Doenças das PlantasRESUMO
The complete genome sequence of Stellaria aquatica virus B (StAVB), a new member of the genus Polerovirus that infects Stellaria aquatica, was determined using high-throughput RNA sequencing with confirmation by Sanger sequencing. The complete StAVB genome (GenBank accession no. OP389993) is 5,900 nucleotide (nt) long with seven open reading frames (ORF0-5 and ORF3a) that encode putative proteins (P0-P5 and P3a) in a similar configuration to that of other typical poleroviruses. Pairwise sequence comparisons with other poleroviruses showed 38-50% nt sequence identity in the complete genome and 13-24%, 36-45%, 7-68%, and 6-50% amino acid sequence identity in (aa), for the P0, P1-2, P3, and P4 protein, respectively. These data, together with the results of phylogenetic analysis, indicate that StAVB should be classified as a new member of the genus Polerovirus, family Solemoviridae.
Assuntos
Luteoviridae , Stellaria , Luteoviridae/genética , Stellaria/genética , Genoma Viral , Filogenia , Doenças das Plantas , Sequenciamento de Nucleotídeos em Larga Escala , Fases de Leitura Aberta , RNA Viral/genéticaRESUMO
The genome of a new potyvirus from a Lamprocapnos spectabilis plant in South Korea was sequenced by high-throughput sequencing and confirmed by Sanger sequencing. The new potyvirus was tentatively named "lamprocapnos virus A" (LaVA); its complete genome contains 9,745 nucleotides, excluding the 3'-terminal poly(A) tail. The LaVA genome structure is similar to that of members of the genus Potyvirus and contains an open reading frame encoding a large putative polyprotein of 3,120 amino acids (aa) with conserved motifs. The complete genome shared 48%-56% nucleotide sequence identity and the polyprotein shared 41%-52% aa sequence identity with those of other potyviruses. These values are below the standard thresholds for potyvirus species demarcation. Phylogenetic analysis based on polyprotein sequences showed that LaVA belongs to the genus Potyvirus. To our knowledge, this is the first report of the complete genome sequence and genome characterization of a potyvirus infecting Lamprocapnos spectabilis.
Assuntos
Genoma Viral , Potyvirus , Potyvirus/genética , Filogenia , RNA Viral/genética , Fases de Leitura Aberta , Poliproteínas/genética , Doenças das PlantasRESUMO
A new member of the genus Alphacarmovirus was detected in Stellaria aquatica using high-throughput RNA sequencing analysis. The complete genome sequence of this new virus isolate, tentatively named "Stellaria aquatica virus A" (StAV-A), comprises 4,017 nucleotides with five predicted open reading frames (ORFs) and has a typical alphacarmovirus genome organization. Pairwise comparison of StAV-A with selected members of family Tombusviridae showed 44-58%, 32-64%, and 19-49% sequence identity for the overall nucleotide sequence, polymerase, and coat protein, respectively. Phylogenetic analysis of polymerase sequences places StAV-A alongside other members of the genus Alphacarmovirus in the family Tombusviridae.
Assuntos
Stellaria , Tombusviridae , Genoma Viral , Stellaria/genética , Filogenia , RNA Viral/genética , Fases de Leitura Aberta , Doenças das PlantasRESUMO
A novel umbra-like virus was identified in arborvitae in South Korea using RNA sequencing (RNA-seq). The virus identified was tentatively named "arborvitae umbra-like virus" (AULV) and contained a 4,300-nucleotide genome organized into four non-structural open reading frames (ORFs). Cloning and Sanger sequencing were used to confirm the viral contig sequence and determine the size of the genome. Genome analysis indicated that ORF2 encodes an RNA-dependent RNA polymerase that is probably expressed through ribosomal frameshifting. ORF3 encodes a putative long-distance movement protein, while the functions of ORFs 1 and 4 are unknown. The virus lacks a coat protein gene. The genome of AULV shares 27.3%-48.4% nucleotide sequence identity with closely related umbraviruses. Phylogenetic analysis based on the complete genome sequences and amino acid sequences of the RNA-dependent RNA polymerase revealed that AULV forms a monophyletic lineage with Guiyang paspalum paspaloides tombus-like virus (GPpTV1). We suggest that AULV is a novel umbra-like virus belonging to the family Tombusviridae.
Assuntos
Thuja , Tombusviridae , Umbridae , Animais , Filogenia , China , República da Coreia , RNA Polimerase Dependente de RNA/genéticaRESUMO
A novel cytorhabdovirus was identified in Daphne odora in South Korea using high-throughput sequencing. The virus, tentatively named "daphne virus 1" (DV1), has a full-length genome sequence of 13,206 nucleotides with a genome organization comparable to that of unsegmented plant rhabdoviruses and contains seven antisense putative genes in the order 3'-leader-N-P'-P-P3-M-G-L-5'-trailer. The coding region of the genome is flanked by a 3' leader and a 5' trailer sequence, 261 and 151 nucleotides long, respectively. The DV1 genome shares 33.74%-57.44% nucleotide sequence identity with other cytorhabdoviruses. The DV1-encoded proteins share the highest amino acid sequence identity with homologues from Asclepias syriaca virus 1. Phylogenetic analysis showed that DV1 clustered with representative cytorhabdoviruses. We propose classifying DV1 in a new species within the genus Cytorhabdovirus, family Rhabdoviridae.
Assuntos
Daphne , Rhabdoviridae , Daphne/genética , Filogenia , RNA Viral/genética , Rhabdoviridae/genética , Genoma Viral , Proteínas Virais/genética , Fases de Leitura Aberta , Nucleotídeos , Doenças das PlantasRESUMO
The common bean (Phaseolus vulgaris; family: Fabaceae) is an economically and nutritionally important food crop worldwide (Ganesan et al. 2017). In 2021, several plants collected from different provinces in South Korea had symptoms of viral infections (e.g., mild yellow-greenish speckling, stunting, crinkling, and deformed leaves). To identify the causal pathogens, total RNA was isolated from pooled leaf tissues from all samples (n = 29) for paired-end high-throughput sequencing (HTS). The cDNA library was constructed after eliminating ribosomal RNA using the TruSeq RNA Sample Prep Kit and then sequenced using the Illumina NovaSeq 6000 platform (Macrogen, Korea). The 297,868,156 paired-end clean reads (150 nt) were de novo assembled using Trinity with default parameters. BLASTx was used for the contig analysis, which revealed the pooled samples were infected with several plant viruses (e.g., turnip mosaic virus, zucchini yellow mosaic virus, cucumber mosaic virus, lily mottle virus). Notably, the assembled contigs included a single viral contig (8,472 nt) comprising the nearly complete KLV genome (HTS mean coverage: 39.46%). Kalanchoe latent virus (KLV; genus: Carlavirus; family: Betaflexiviridae) has been detected in Kalanchoë blossfeldiana (Hearon 1982), Chenopodium quinoa (Dinesen et al. 2009), and Graptopetalum paraguayense (Sorrentino et al. 2017). The sequence was most similar (96.28% nucleotide identity; 99% query coverage) to KLV isolate DSMZ PV-0290 (GenBank: OP525283) from Denmark. The contig sequence was validated via reverse transcription-polymerase chain reaction (RT-PCR) using total RNA extracted from the 29 individually stored samples and nine primer sets specific for the KLV contig. All nine contig-specific overlapping fragments were amplified from only a P. vulgaris plant with mild yellowing mosaic symptoms collected on July 6, 2021, in Jeongseon County, South Korea. Additionally, 5' and 3' rapid amplification of cDNA ends (RACE)-specific primers were designed for the KLV contig sequence to determine the terminal ends of the genome of the South Korean KLV isolate using the 5'/3' RACE System (Invitrogen, Carlsbad, CA, USA). All of the amplified and overlapping fragments were cloned into the RBC T&A Cloning Vector (RBC Bioscience, Taipei, Taiwan) and sequenced using the Sanger method. The obtained full-length genomic sequence of the KLV isolate (KLV-SK22) was 8,517 nt long and was deposited in GenBank OQ718816. According to the BLASTn analysis, KLV-SK22 was highly similar (96.30% sequence identity; 100% query coverage) to the DSMZ PV-0290 isolate. Phylogenetic trees constructed on the basis of coat protein and RNA-dependent RNA polymerase amino acid sequences revealed that KLV-SK22 is closely related to the DSMZ PV-0290 and PV-0290B isolates from Denmark, respectively. At the genome and gene levels, the individual sequence identities between the carlaviruses and other KLV isolates were 96.29% to 100% (Adams et al. 2004). Additionally, an RT-PCR analysis using detection primers specific for KLV-SK22 did not detect KLV in 15 samples (P. vulgaris = 3, Glycine max = 8, Pueraria montana = 2, Trifolium repens = 1, and Vigna angularis = 1) randomly collected from different regions in South Korea. Based on these results, KLV infection may not be widespread at this time in South Korea. To the best of our knowledge, this is the first report of KLV in P. vulgaris in South Korea or elsewhere. Our findings will aid future research on the epidemiology and long-term management of KLV-related diseases.
RESUMO
The complete genome sequence of a putative new virus isolate, provisionally named "Fagopyrum esculentum endornavirus 2" (FeEV2), is 15,706 nucleotides long with a single, large open reading frame and a typical endornavirus genome organization. FeEV2 shares 19.4%-22.1% nucleotide sequence identity with other known endornavirus genome sequences. The putative polyprotein, RNA-dependent RNA polymerase (RdRp), helicase, and glycosyltransferase (GT) share 10.6%-24.3%, 30.4%-66.1%, 16.3%-45.7%, and 10.1%-21.6% amino acid sequence identity, respectively, with the homologous sequenced proteins from known endornaviruses. This suggests that it is a member of a new, distinct species. Phylogenetic analysis of RdRp sequences places FeEV2 with other Alphaendornavirus genus members (family Endornaviridae). This is the first report of the complete genome sequence of FeEV2, which was isolated from Fagopyrum esculentum in South Korea.
Assuntos
Fagopyrum , Vírus de RNA , Fagopyrum/genética , Genoma Viral , Fases de Leitura Aberta , Filogenia , RNA Viral/genética , RNA Polimerase Dependente de RNA , Proteínas Virais/genéticaRESUMO
Here, we report the complete genome sequence of a novel polerovirus, "Plantago asiatica virus A" (PlaVA), detected in Plantago asiatica using high-throughput RNA sequencing and validated by Sanger sequencing. The complete PlaVA genome contains 5,881 nucleotides and has seven open reading frames (ORF0-5 and ORF3a) encoding putative proteins (P0-5 and P3a, respectively) in an arrangement that is similar to that of typical Polerovirus members. Pairwise sequence comparisons revealed that P0 to P5 encoded by PlaVA had the highest sequence identity (25.48%-79.21%) to the corresponding proteins of previously reported poleroviruses. A phylogenetic analysis using the PlaVA P1-2 and P3 amino acid sequences and those of members of the family Solemoviridae (formerly Luteoviridae) indicated that although PlaVA belongs to the genus Polerovirus, it does not represent a known species. Consequently, PlaVA should be considered a member of a new species within the genus Polerovirus.
Assuntos
Luteoviridae , Plantago , Genoma Viral , Sequenciamento de Nucleotídeos em Larga Escala , Luteoviridae/genética , Fases de Leitura Aberta , Filogenia , Doenças das Plantas , RNA Viral/genéticaRESUMO
The complete genome sequence of a new caulimovirus in Pueraria montana was determined using high-throughput sequencing. The 7,572 nucleotide genome of pueraria virus A (PVA) contains genes that encode a movement protein, an aphid transmission factor, a virion-associated protein, a coat protein, a protease + reverse transcriptase + ribonuclease H, and a transactivator/viroplasmin protein, as well as two intergenic regions, which are all common features of members of the genus Caulimovirus. A sequence alignment revealed that the complete genome of PVA shares 66.82% nucleotide sequence identity with strawberry vein banding virus (GenBank accession no. KX249738.1). The results of phylogenetic analysis and the observation that the nucleotide sequence of the polymerase coding region differed by more than 20% indicated that PVA is a member of a new species the genus Caulimovirus, family Caulimoviridae.
Assuntos
Caulimoviridae , Pueraria , Vírus não Classificados , Caulimoviridae/genética , Caulimovirus , Genoma Viral , Sequenciamento de Nucleotídeos em Larga Escala , Fases de Leitura Aberta , Filogenia , Doenças das Plantas , Vírus não Classificados/genéticaRESUMO
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.
Assuntos
Luteoviridae , Doenças das Plantas , Sequência de Bases , Genoma Viral , Sequenciamento de Nucleotídeos em Larga Escala , Luteoviridae/genética , Fases de Leitura Aberta , Filogenia , RNA Viral/genética , Proteínas Virais/genéticaRESUMO
A new virus, tentatively named "kudzu virus D" (KuVD) was discovered in kudzu (Pueraria montana var. lobata) in South Korea. Its complete genome comprises 7,922 nucleotides, excluding the poly(A) tail, and contains five open reading frames (ORFs) encoding, from 5' to 3', a replicase (ORF1), three triple gene block proteins TGB1-3 (ORF2-ORF4), and a coat protein (ORF5). This genome organization is typical of members of the subfamily Quinvirinae of the family Betaflexiviridae. Pairwise alignment analysis revealed that the nucleotide sequences of the replicase and coat protein of KuVD were 12.13-54.46% and 24.03-50.67% identical, respectively, to those of other members of the family Betaflexiviridae. These values are far below the current species ICTV demarcation threshold. Consequently, KuVD should be considered a member of a new species in the subfamily Quinvirinae.
Assuntos
Flexiviridae , Pueraria , Flexiviridae/genética , Genoma Viral , Fases de Leitura Aberta , Filogenia , Doenças das PlantasRESUMO
The genome of a novel virus identified in Cnidium officinale is composed of a monopartite ssRNA of 16,755 nucleotides that shares 68.73% (query coverage, 20%) sequence identity with carrot yellow leaf virus (CYLV, accession no. FJ869862.1). It contains 11 putative open reading frames and has an organization typical of closteroviruses. It shares 30-50% nucleotide sequence identity with other closteroviruses. The heat shock protein 70-like protein (HSP70), putative RNA-dependent RNA polymerase (RdRp), and coat protein (CP) show 39-66%, 16-60%, and 24-41% amino acid sequence identity, respectively, to the homologous proteins of previously identified closteroviruses. Molecular and HSP70-based phylogenetic analysis of the genome and encoded protein sequences suggested that this virus is a novel member of the genus Closterovirus in the family Closteroviridae, which we have tentatively named "cnidium closterovirus 1" (CnClV1).
Assuntos
Closterovirus , Closterovirus/genética , Cnidium , Genoma Viral , Fases de Leitura Aberta , Filogenia , RNA Viral/genéticaRESUMO
The complete genomic sequence of a plant rhabdovirus that was identified in Cnidium officinale in Yeongyang-dun, South Korea, is reported here. The virus, tentatively named "cnidium virus 1" (CnV1), has a negative-sense RNA genome of ~ 14 kb, and its organization most closely resembles that of unsegmented plant rhabdoviruses, containing six antisense open reading frames (ORFs) in the order 3'-N-P-P3-M-G-L-5'. Intergenic regions containing conserved sequences separate the genes. The genome of CnV1 is 37.8-56% identical in its complete nucleotide sequence to betanucleorhabdoviruses and other related rhabdoviruses. Therefore, based on the sequence similarity criteria for species demarcation, its genome organization, and its phylogenetic position, CnV1 should be classified as a new member of the genus Betanucleorhabdovirus in the family Rhabdoviridae. CnV1 is the first rhabdovirus found in C. officinale.
Assuntos
Cnidium , Rhabdoviridae , Genoma Viral , Fases de Leitura Aberta , Filogenia , Doenças das Plantas , RNA Viral/genética , Rhabdoviridae/genéticaRESUMO
In this work, two new turnip mosaic virus (TuMV) strains (Canola-12 and Canola-14) overcoming resistance in canola (Brassica napus) were isolated from a B. napus sample that showed typical TuMV-like symptoms and was collected in the city of Gimcheon, South Korea, in 2020. The complete genome sequence was determined and an infectious clone was made for each isolate. Phylogenetic analysis indicated that the strains isolated from canola belonged to the World-B group. Both infectious clones, which used 35S and T7 promoters to drive expression, induced systemic symptoms in Nicotiana benthamiana and B. napus. To our knowledge, this is the first report of TuMV infecting B. napus in South Korea.
Assuntos
Brassica napus , Potyvirus , Células Clonais , DNA Complementar/genética , Filogenia , Doenças das Plantas , Potyvirus/genéticaRESUMO
Three cotton leafroll dwarf virus (CLRDV; genus Polerovirus, family Solemoviridae) genotypes have recently been identified (Tabassum et al., 2021; Ramos-Sobrinho et al., 2021). This virus is widespread in the United States (Thiessen et al., 2020; Aboughanem-Sabanadzovic et al., 2019; Tabassum et al., 2020) and has also been reported to infect chickpea (Cicer arietinum) in Uzbekistan (Kumari et al., 2020). As well, CLRDV was detected from 23 weed species (16 families), including Hibiscus sabdariffa (Sedhain et al., 2021, Hagan et al., 2019). From June to September 2019, virus-like symptoms, including mild leaf stunting, crinkling, and deformation, were observed in multiple plants (n=14) in several provinces of South Korea (e-Xtra Table. 1). To characterize the associated viruses, pooled leaf tissues from all 14 samples were used for total RNA isolation, followed by paired-end high-throughput sequencing (HTS) on the Illumina NovaSeq 6000 platform (Macrogen, South Korea). A total of 614,424,952 trimmed and high-quality reads were assembled into 506,024 contigs using Trinity de novo transcriptome assembly. The resulting contigs were compared with viral sequences in the GenBank database using BLASTx analysis. Several viral contigs were identified, including cucumber mosaic virus, apple stem pitting virus, apple stem grooving virus, cherry virus A, and CLRDV. The CLRDV contig of 5,800 nucleotides (nt) with an average coverage of 307x shared 92.1% identity (query coverage: 99%) with the CLRDV isolate CN-S5 (KX588248). To confirm CLRDV infection and to obtain its complete genome sequence, total RNA was extracted from each of the 14 samples and used for reverse transcription (RT)-PCR with six overlapping sets of primers designed from the HTS contig (e-Xtra Table. 2). The expected product sizes were obtained only for the Hibiscus syriacus L. (family: Malvaceae) sample showing foliar mild vein clearing symptoms on the leaves (e-Xtra Fig.1). All RT-PCR products were cloned using the RBC TA Cloning vector (Taipei, Taiwan) and at least five positive clones per cloned DNA fragment were sequenced. The 5ï¢ and 3ï¢ termini sequences were determined as described previously (Zhao et al. 2016). The complete genome of CLRDV isolate SK (OK073299) was determined to be 5,862 nt and it shared 89-91% complete genome identity with 12 other CLRDV isolates based on pairwise comparisons (e-Xtra Table. 3). Maximum likelihood phylogenetic analysis based on the complete genome and P3-CP aa sequences showed that CLRDV-SK is more closely related CN-S5 (e-Xtra Fig. 2). In the fall of 2021, additional H. syriacus samples (n=18) with mild chlorosis, blistering and crinkling symptoms were collected from 2 provinces of South Korea and tested by RT-PCR using the primers: CLRDV-SK-101-120 For & CLRDV-SK-1021-1040 Rev targeting a region of the ORF0. Two of 18 samples (11.1%) tested positive for CLRDV. The 16 negative samples only showed symptoms of mild yellowing. The RT-PCR products were cloned and sequenced. In pairwise comparisons, the obtained sequences (OM339522-23) were 95.85% and 96.06% identical to the corresponding sequences of CLRDV isolate SK. This is the first report of CLRDV occurrence in H. syriacus in South Korea to the best of our knowledge. Our findings will assist further studies on the epidemiology and sustainable management of diseases caused by CLRDV. Acknowledgments This work was supported by IPET (Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries; Project No. AGC1762111), Ministry of Agriculture, Food and Rural Affairs, Republic of Korea. References Tabassum, A., et al., 2021. PloS One. 16: e0252523 Ramos-Sobrinho, R., et al., 2021. Viruses. 13:2230 Thiessen, L.D., et al. 2020. Plant Dis. 104:3275 Aboughanem-Sabanadzovic, N., et al. 2019. Plant Dis. 103:1798 Tabassum, A., et al. 2020. Microbiol. Res. Announce. 9:e00812-20 Kumari, S.G., et al. 2021. Plant Dis. 104:2532 Sedhain, N.P., et al. 2021. Crop protection 144:105604 Hagan, A., er al. 2019. Alabama Cooperative Extension System. ANR:2539 Zhao, F., et al. 2016. Arch. Virol. 161:2047 Conflict of interest The authors declare that they have no conflict of interest.