Molecular and biological characterization of a newly identified virus representing a novel taxon of Alphaflexiviridae infecting different accessions of seashore paspalum, a turfgrass, widely grown in the United States

Molecular and biological characterization of a newly identified virus representing a novel taxon of Alphaflexiviridae infecting different accessions of seashore paspalum, a turfgrass, widely grown in the United States | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Molecular and biological characterization of a newly identified virus representing a novel taxon of Alphaflexiviridae infecting different accessions of seashore paspalum, a turfgrass, widely grown in the United States Sayanta Bera, Taylor F. Schulden, Xiaojun Hu, Peter Abrahamian, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8745768/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background Seashore paspalum ( Paspalum vaginatum ), a widely cultivated turfgrass in the United States, is valued for its salinity tolerance and environmental adaptability. Despite its economic importance, viral infections in paspalum remain poorly characterized. This study reports the discovery and characterization of a novel virus infecting paspalum, tentatively named Paspalum latent virus (PaLV), representing a new taxon within the family Alphaflexiviridae . Methods High-throughput sequencing, RACE PCR, and Sanger sequencing were used to obtain complete genome sequences of PaLV from multiple paspalum accessions. Phylogenetic analysis was performed using replicase protein sequences. Structural modeling of the coat protein (CP) was conducted using AlphaFold2 and TM-align. RT-PCR assays were developed for virus detection and host range studies. Genetic diversity and population structure were assessed using CP sequences from 11 PaLV isolates. Results PaLV possesses a 6,995 nt long genome with five ORFs and lacks the AlkB domain in its polymerase gene . The genome exhibits unique features, including overlapping start-stop codons between ORF4 and ORF5 and a second in-frame AUG codon in the CP region. Phylogenetic analysis placed PaLV in a distinct clade clearly separated from Potexvirus and Lolavirus . Structural modeling revealed conserved CP domains despite low sequence identity. Genetic analysis indicated low diversity and strong purifying selection, with limited spatial structure. An RT-PCR detection assay was developed to detect PaLV in paspalum accessions which was detected at an incidence rate of 90% . Mechanical transmission resulted in infection of Zea mays , Sorghum spp. , Setaria italica , and Lolium multiflorum , without visible symptoms. Conclusions PaLV represents a novel viral species and likely a new tentatively named genus, Paspalovirus , within the family Alphaflexiviridae . Its latent infection profile and broad host range underscore the need for routine molecular diagnostics in turfgrass management. The RT-PCR assay developed here provides a reliable tool for PaLV detection, and further studies are warranted to assess its impact on plant physiology and breeding. Lolavirus turfgrass evolution germplasm Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Turfgrasses play a vital role in environmental protection and human well-being [ 1 ]. Seashore paspalum ( Paspalum vaginatum Sw. ) , a warm-season perennial turfgrass, has attracted attention due to its salinity tolerance and adaptability to coastal environments in tropical and subtropical regions [ 2 , 3 ]. As a result, seashore paspalum is a popular choice for sports fields in coastal regions where freshwater availability is an issue and saltwater irrigation results in tremendous cost savings [ 4 ]. Beyond its functional benefits, seashore paspalum also contributes to environmental sustainability by mitigating the urban heat island effect, providing forage and aesthetic value, and generating economic and social benefits. The species is also reportedly tolerant to flooding and low oxygen conditions [ 1 , 5 , 6 ]. Due to these numerous benefits, the cultivation area of Paspalum has expanded beyond coastal areas; however, in new environments, plants are exposed to many unknown pathogens, increasing the risk of disease damage. Seashore paspalum, similar to other turfgrasses, is propagated through clonal methods, resulting in genetic uniformity [ 7 ]. As a result, viruses are easily spread across the propagated plant material and introduced into new areas [ 7 , 8 ]. Notable viruses that have been identified in Paspalum , under natural or experimental conditions, include Paspalum striate mosaic virus (PSMV, Geminiviridae ) [ 9 , 10 ], sugarcane mosaic virus (SCMV, Potyviridae ) [ 11 ], sugarcane streak Réunion virus (SSRV, Geminiviridae ) [ 12 ], Chloris striate mosaic virus (CSMV, Geminiviridae ) [ 9 ], Paspalum dilatatum striate mosaic virus (PDSMV, Geminiviridae ) [ 9 ], and barley yellow dwarf virus (BYDV, Tombusviridae ) [ 13 ]. In recent years, high-throughput sequencing (HTS) has emerged as a transformative technology in the field of virology, particularly in the realm of viral discovery and diagnostics [ 14 , 15 ]. This approach has led to the identification of numerous novel viruses in diverse biological samples, including grasses, weeds, trees, and food crops [ 16 – 20 ]. Therefore, in this study we leveraged HTS to investigate the virome of Paspalum germplasm used in breeding programs. For this purpose, 27 P. vaginatum and three P. distichum (a sister species of P. vaginatum ) accessions were obtained from the USDA National Plant Germplasm System (NPGS) collection and maintained at the University of Georgia (UGA), Griffin and Athens campuses. Our results include the first report of a virus representing a novel taxon within the Alphaflexiviride that infects both P. vaginatum and P. distichum . Confirmation of putative viral sequences was obtained by RT-PCR and genome terminals were completed with 5' RACE. Based on phylogenetic analysis and genomic structure, we propose that this novel virus, tentatively named Paspalovirus paspali (Paspalum latent virus, PaLV) is a new virus, in the family Alphaflexiviridae . Here, we report the identification of PaLV, its biological and molecular characterization, phylogenetic relationships, genetic diversity and population structure, and PaLV screening of P. vaginatum and P. distichum germplasm maintained in greenhouses at UGA. Material and Methods Plant Material and Growth Paspalum plants, obtained initially from the USDA-NPGS collection, Griffin, were maintained in greenhouses at the UGA Griffin and Athens campuses in 4x4 pots. The soil was a 1:1 mixture of sand and Miracle-Gro potting mix. Plants were fertilized once every two weeks with Osmocote Plus. Total Plant RNA Extraction Total RNA from the Paspalum accession PI 509022 leaf tissue was extracted using a RNeasy Plant Mini Kit (Qiagen, Hilden, Germany), and an RNase-Free DNase Set (Qiagen) was used for DNA digestion. The manufacturer’s recommended protocol was used without modifications. For accessions HI10 and Spence, papillae were peeled from the adaxial surface of leaves (second fully open, developed leaf on growing stolon segment) from plants grown under freshwater (0 mM NaCl) and salt stress (200 mM NaCl) irrigation for six weeks. Strings of papillae were collected in 2 mL tubes resting on a liquid nitrogen bath to maintain RNA integrity, and frozen tissues were ground using a TissueLyser II bead mill (Qiagen). The ground tissue was homogenized in 1 mL of TRIzol reagent (Invitrogen, Waltham, MA) per 100 mg of tissue, followed by phase separation after adding 200 µL of chloroform. The RNA-containing aqueous phase was carefully transferred to a 2 mL nuclease-free tube and mixed with an equal volume of 100% ethanol. The resulting RNA-ethanol mixture was loaded onto a Zymo-Spin IC column from the Zymo RNA Clean & Concentrator-5 kit (Zymo Research, Irvine, CA), and RNA clean-up was performed following the manufacturer’s instructions. RNA was eluted in 10 µL nuclease-free water and RNA concentration was determined using a NanoDrop spectrophotometer, and the quality was assessed via electrophoresis on a 1% Tris-Borate-EDTA (TBE) agarose gel. HTS and Virus Identification The RNA extracts from leaves of accession PI 509022 at USDA APHIS-PPQ, Beltsville, and from adaxial leaf papillae from accessions HI 10 and Spence at the University of Georgia, Athens, were subjected to HTS at their respective place of extraction. For HTS of RNA from PI 509022, a single-indexed ribosomal-RNA-depleted cDNA library was prepared using the TruSeq® Stranded Total RNA Library Prep Plant kit (Illumina, San Diego, CA) and sequenced on an Illumina NextSeq 500 platform, generating 19,815,890 single-end, 75-bp reads. For HTS of RNA from accessions HI 10 and Spence, barcoded stranded mRNA-seq libraries were generated from 500 ng of RNA with the KAPA Stranded mRNA-Seq Kit (Roche, Basel, Switzerland) according to the manufacturer's instructions, using half-reactions for all steps except the final library amplification. The concentration of the libraries was measured using a Qubit Fluorometer with the Qubit 1X dsDNA High Sensitivity Assay Kit (Invitrogen). Subsequently, 30 ng of each library was pooled into a larger set of 24 leaf and peels libraries and loaded onto a single flow cell of an Illumina NextSeq 2000 for paired end (PE) 150 bp sequencing at the Georgia Genomics and Bioinformatics Core (GGBC) at UGA. Reads from HTS were assembled using the de novo assembly tool in PhytoPipe (2023) [ 21 ] and compared with viral pathogen databases. Briefly, raw reads were filtered and trimmed using Trimmomatic (v.0.39) [ 22 ]. Then, contigs were assembled using Trinity (v2.8.6) [ 23 ] and compared to the NCBI Viral Reference Database (Jan 2022) by BLASTn search [ 24 ] and to the Reference Viral Database protein database (v22)[ 25 ] by Diamond blastx search [ 26 ]. Default parameters were used in all programs. 5' RACE Rapid amplification of cDNA ends with the SMARTer® RACE 5′/3′ Kit (Takara) was used to complete the 5′ end of the ssRNA segment with the RACE primer – 5' - GGACCACTGTGGCGTAGAGGATGTCGAGGT-3'; the 5' end of the primer was linked to the adapter sequence provided by the company. All PCR fragments were sequenced by direct Sanger sequencing in both directions, and sequences were aligned to the reference HTS-derived contigs using Geneious (v11.0.3). RT-PCR Validation of HTS and Screening of Germplasm A one-step RT-PCR protocol was developed for detection of PaLV using the SuperScript™ III One-Step RT-PCR System with Platinum® Taq DNA Polymerase (Invitrogen). Primer pairs ORF1.FP: 5′-AAACAAGTGGAATTTCACGGG-3′ and ORF1.RP: 5′-CTGCTCTGGTGAGAAGATATCG-3′, were designed to amplify a 605 bp amplicon from the replicase gene. Cycling parameters were used following the manufacturer's guidelines except for the 62°C annealing temperature. Host Range Studies After confirmation of PaLV infection in Paspalum through RT-PCR, infected plant tissues were freshly ground in 0.01M potassium phosphate buffer (pH 7) for mechanical inoculation. All plants were sprayed with carborundum, an abrasive, before rub-inoculation. Mechanical inoculation of sap was conducted on nine different plant species: Hordeum vulgare, Triticum aestivum, Zea mays, Sorghum spp., Dactylis glomerata, Setaria italica, Lolium multiflorum, Miscanthus sacchariflorus , and Avena sativa . Three biological replicates were used for each species. All bioassay plants were kept in controlled-temperature greenhouse conditions at 26℃ ± 2℃ with 16 h light and 8 h dark. Two plant species, H. vulgare , and T. aestivum were assayed in a growth chamber maintained at 16℃ with 16 h light and 8 h dark. Sequence, Phylogenetic, and Recombination Analysis The viral sequences were analyzed by BLASTn ( https://blast.ncbi.nlm.nih.gov/Blast.cgi ) with default parameters. The putative proteins and potential open reading frames (ORFs) were determined using ORFfinder ( https://www.ncbi.nlm.nih.gov/orffinder/ ) and subsequent blastp annotation by using the non-redundant protein sequences database. Pairwise comparisons between related viral ORFs were performed with Sequence Demarcation Tool (SDT) v1.2 to calculate the identity percentage for each ORF [ 27 ]. Conserved domains within these proteins were identified using the Conserved Domain Database (CDD) [ 28 ]. The coat protein (CP) sequences were modeled using AlphaFold2[ 29 ]. PaLV CP (CP PaLV ) was superimposed on the Lolium latent virus (CP LoLV ) using TM-align [ 30 ]. Structural similarity between CP PaLV and CP LoLV was further evaluated based on TM scores, using 0.5 as a cut-off. Superimposed images were exported from TM-align and loaded into Chimera X [ 31 ] for further processing. Evolutionary relationship of PaLV with representative species from the genera, Lolavirus , Potexvirus , Mandarivirus , and Allexivirus in the Alphaflexiviridae family were included in a maximum-likelihood phylogenetic tree based on amino acid sequences of replicase using the LG with gamma distribution and frequency substitution model previously inferred using jModelTest in MEGA 11 [ 32 ]. The phylogenic analysis was performed with 1,000 bootstrap replicates and potato virus T as an outgroup. The resulting phylogenetic tree was visualized and formatted using FigTree v1.4.4 ( http://tree.bio.ed.ac.uk/software/figtree/ ). The Recombination Detection Program V.5 (RDP5) program was used to explore the occurrence of recombination events in full-length viral genome sequences. Various methods implemented in RDP5 [ 33 ] including RDP, SisterScan, Bootscan, Chimaera, GeneConv, MaxChi, and 3Seq algorithms, were used. Any recombinant events with P < 0.001 predicted by four of the seven prediction methods were considered reliable and used in the analysis. Genetic diversity The genetic diversity of the PaLV population was analyzed using 11 PaLV isolates obtained from different Paspalum accessions from the UGA collection. Genetic diversity was calculated based on the complete CP gene nucleotide sequences. The primer set designed to amplify the CP region of PaLV was developed according to the consensus sequence of three PaLV isolates obtained through HTS (CP_F: 5′-GATCGGAAGCCTCAGTTGTG-3′; CP_R: 5′-CGGTGGCCAGGGTAGATTAA-3′). The CP nucleotide sequence of 11 PaLV isolates was aligned using the Muscle program implemented in MEGA 11 and the mean genetic diversity for the entire population was calculated. The average number of non-synonymous (d N ) and synonymous (d S ) nucleotide substitutions per site and the dN/dS ratio as an estimate of the CP's selection pressure were computed using the DnaSP v.5 software [ 34 ]. The Wright’s fixation index statistic F ST [ 35 ] was calculated using DnaSP v.5. Frequent gene flow is considered to occur when F ST < 0.33. Results Genome Structure of PaLV High-throughput sequencing and 5′ RACE revealed the complete genome of PaLV to be 6,995 nucleotides (nt) in length, excluding the poly(A) tail. The genome comprised five open reading frames (ORFs), flanked by an 81 nt 5′ untranslated region (UTR) and an 86 nt 3′ UTR. Pairwise comparisons of PaLV ORFs with representative Alphaflexiviridae viruses showed highest identity with Lolium latent virus (LoLV), ranging from 49–57% at the nucleotide level and 26–55% at the amino acid level (Table 1). ORF1 encoded a 173-kDa replicase protein containing conserved domains: Methyltransferase (aa 39–329), Viral RNA helicase (aa 768–997), and RNA-dependent RNA polymerase (aa 1178–1474) (Fig. 1 A). The AlkB domain, which is present in LoLV, was absent in ORF 1 of PaLV. ORFs 2–4 encoded triple gene block (TGB) proteins: TGB1 (30.4 kDa), TGB2 (13.5 kDa), and TGB3 (7.8 kDa). These proteins have been shown to play a crucial role in virus movement [ 36 ]. A putative octanucleotide promoter sequence for sgRNA1 (CUUAAGUU) was identified 21 nt upstream of ORF2, overlapping the stop codon of ORF1 (Fig. 1 B). ORF5 encoded the coat protein (CP) of 35.4 kDa with a start codon in-frame and immediately followed the stop codon of ORF4, with no intergenic region (Fig. 1 C). A putative sgRNA2 promoter sequence (GUUAAGUU) was located 14 nt upstream of ORF5, within ORF4. ORF6, present in LoLV and other tentative lolaviruses such as Weeping alkaligrass lolavirus 1 (WaLoV1), that overlaps the 3′ end of ORF 5, was not identified in PaLV or the other tentative lola-like members, such as Foxtail millet lolavirus 1 (FmLoV1) [ 37 ] (Fig. 1 A). Comparison of CP PaLV with CP LoLV Subcellular localization prediction using Plant-mSubP indicated that CP PaLV was likely localized in cytoplasm or peroxisomes, while CP LoLV and CP PVX were predicted to localize in plastids. Subsequently, the protein structure of CP PaLV and CP LoLV was modeled to compare their structural relatedness. For this purpose, both amino acid sequences were subjected to Alphafold2 protein structure predictions (Fig. 2 ). The structure of CP PaLV was predicted with a lower confidence level than that of CP LoLV (Fig. 2 ). The structural modeling of both CPs revealed the presence of three domains: i) flexible N-terminal, ii) the core, and iii) C-terminal extensions. The N-terminus of PaLV does not contain a proteolytic cleavage site for chloroplast transit peptide that yields a smaller CP isoform in LoLV. However, a smaller ORF of 28kDa was predicted within the CP ORF. The core and C-terminal regions of the CP play a crucial role in protein-protein and protein-RNA interactions and have been found to show the highest degree of structural conservation in Alphaflexiviridae [ 38 ]. Consequently, a comparative analysis of the core and C-terminal domains (151–300 and 106–280 aa in CP PaLV and CP LoLV , respectively) of both CPs was conducted. As predicted, a high TM score, indicative of high structural similarity, was obtained upon the superimposition of the core and C-terminal regions of both CPs, which consists of conserved α-helical secondary structures (Fig. 2 ). Phylogenetic Analysis Maximum-likelihood phylogenetic analysis based on replicase amino acid sequences placed PaLV in a distinct clade clearly separated from the Potexvirus and Lolavirus genera. The analysis also included putative viral sequences obtained through data mining and submitted as Third-Party Annotation (TPA), such as Saltwater paspalum lolavirus 1 (SpLoV1, synonym PaLV [ 37 ], FmLoV1, a Lola-like virus from Maize (Lola maydis), and WaLoV1. Their inclusion further supports the formation of this novel group within the Alphaflexiviridae family. All PaLV isolates along with SpLoV1formed a unique and well-supported clade (Fig. 3 ). Notably, SpLoV1 shared 97% nucleotide identity with PaLV, which indicates the same viral species. FmLoV1 and Lola maydis sequences represent two potential new members within the PaLV clade. Whereas WaLoV1 appeared to cluster within the Lolavirus clade. No recombination events were detected in PaLV genomes using any of the algorithms implemented in RDP5. Genetic Diversity of PaLV Population Analysis of eleven CP sequences from PaLV isolates revealed two distinct clusters: Cluster A (primarily U.S. origin) and Cluster B (primarily rest of world). F ST value was 0.124, indicating low spatial genetic structure. To ascertain whether the low diversity could be attributed to selection, the dN/dS ratio was estimated, yielding a value of 0.042, suggesting the presence of strong purifying (negative) selection on CP. This finding is further corroborated by the observation of low genetic diversity (0.031 ± 0.004) of CP. Host Range Studies All inoculated (virus and mock) plants were examined by RT-PCR after nine indicator plants representing nine plant genera were subjected to mechanical inoculation with leaf extract from PaLV-infected plants. All inoculated plants of the nine assayed genera were monitored over a period of 21 days post-inoculation (dpi) for symptom development. (Supp Table 1). PaLV amplicons of the expected size were obtained with RT-PCR in: Zea mays , Sorghum spp., Setaria italica , and Lolium multiflorum . Interestingly, T. aestivum plants maintained at 16℃ were also susceptible to PaLV infection based on RT-PCR. The infection of the aforementioned species indicates they are hosts of PaLV. On the contrary, no amplicons were amplified from the remaining inoculated species: Hordeum vulgare , Avena sativa , Dactylis glomerata , and Miscanthus sacchariflorus . None of the experimental hosts showed any symptoms of virus infection. Similarly, no symptoms were also observed in infected seashore paspalum germplasm. No symptoms were observed in mock-inoculated plants. In contrast, LoLV infections in Lolium plants were reported either to be asymptomatic or showing mild chlorotic streaking on the leaves[ 39 ] and in Nicotiana benthamiana plants systemic symptoms such as mosaic and vein netting showed at 15 dpi [ 40 ]. Screening of Paspalum Germplasm A one-step RT-PCR assay targeting ORF1 was developed. A primer pair was designed to target the RNA-dependent RNA polymerase domain in ORF1, amplifying a region of 605 bp. Thirty different Paspalum accessions maintained at UGA were analyzed. Two samples of butterfly bush were used as a negative control, and a PaLV-positive plant as a positive control. The RT-PCR results revealed the presence of PaLV infection in 27 out of 30 accessions (Supp Table 2), with an incidence of 90% in the tested Paspalum accessions. Notably, none of the negative controls showed the presence of PaLV, indicating the specificity of the primer pair. Discussion In this study, we report the molecular and biological characterization of Paspalum latent virus (PaLV), a novel virus infecting seashore paspalum and representing a new taxon within the family Alphaflexiviridae . The complete genome structure, phylogenetic placement, host range, and population diversity of PaLV were investigated, revealing several unique features that distinguish it from known members of the Lolavirus genus. Although PaLV shares moderate sequence identity with Lolium latent virus (LoLV), its genome organization diverges significantly. The absence of the AlkB domain in the replicase, absence of a proteolytic cleavage site within the CP, and the overlapping start-stop codon arrangement between ORF4 and ORF5 suggest distinct evolutionary adaptations. This is a notable divergence from LoLV. Sister members of the PaLV clade, FmLoV1 and Lola maydis, also lack the AlkB domain within the replicase. On the other hand, WaLoV1, which clusters with LoLV contains the AlkB domain. The AlkB domain is speculated to play a crucial role in maintaining the stability of RNA genomes, which are susceptible to methylation due to pesticide applications [ 39 , 41 ]. This speculation led to the hypothesis that the acquisition of the AlkB domain is a relatively recent development in the evolution of viruses within the Alphaflexiviridae family [ 39 , 41 ]. Furthermore, PaLV lacks another signature feature of lolaviruses, ORF6, which is present also in WaLoV1 PaLV shows similar protein sizes for the various ORFs to FmLoV1 rather than LoLV and WaLoV1[ 37 , 39 ]. Overall, the absence and presence of these signature features across both clades further support the divergence across both taxa. As a result, this supports our hypothesis that PaLV represents a new genus, tentatively named ‘ Paspalovirus ’. Based on the ICTV demarcation criteria in the family Alphaflexiviridae , viruses are separated into different genera when their coat protein or polymerase genes share less than about 45% amino acid identity. Whereas, viruses are considered distinct species if they share less than 72% nucleotide identity or less than 80% amino acid identity in either their coat protein (CP) or replicase genes. In this study, PaLV replicase showed less than 45% amino acid identity with LoLV, which is below the genus demarcation threshold and the CP amino acid identity was below 40%. Further, there is incongruency in other features, such as different genome organization and putative protein sizes, in placing PaLV within the Lolavirus or other established genera. Despite low amino acid identity between PaLV and LoLV CPs, structural modeling revealed some conserved core and C-terminal domains. These domains are analogous to the crystal structures of CPs of PapMV and Pepino mosaic virus from the Potexvirus genus [ 38 , 42 ]. The N- and C-terminal extensions play a crucial role in the polymerization of CP [ 38 ]. The core and C-terminal regions of the CP play a crucial role in protein-protein and protein-RNA interactions and have been found to show the highest degree of structural conservation in Alphaflexiviridae [ 38 ]. These regions are critical for virion assembly and RNA binding, suggesting functional conservation. Interestingly, subcellular localization predictions indicated that PaLV CP may localize to cytoplasm or peroxisomes, unlike LoLV and PVX CPs, which preferentially target plastids. This divergence coupled with a very different N-terminus sequence within PaLV CP compared to LoLV may influence host interactions and symptom expression. For instance, tentative members FmLoV1 and Lola maydis have several internal smaller ORF within the CP. Further experimentation beyond the scope of this work is needed to validate whether one or more CP isoforms exist for paspaloviruses. PaLV was detected in multiple grass species, including Zea mays , Sorghum spp. , Setaria italica , and Lolium multiflorum , indicating a broad host range. However, no visible symptoms were observed in any infected plants, including paspalum accessions. This latent infection profile complicates field diagnostics and may contribute to unnoticed virus spread through vegetative propagation. In contrast, LoLV infections in Lolium plants were reported either to be asymptomatic or showing mild chlorotic streaking on the leaves [ 39 ] and in experimental plants systemic symptoms such as mosaic and vein netting showed at 15 dpi [ 40 ]. This indicates that while PaLV shares similarity to LoLV, PaLV does not show severe or adverse effects due to the lack of virulence factors which can allow the virus to go undetected in some grasses. Analysis of 11 PaLV isolates revealed low genetic diversity and strong purifying selection on the CP gene. A F ST of 0.124 indicated a low degree of spatial structure and genetic differences between the population [ 43 , 44 ]. These results are consistent with the expectation of CP sequence conservation given the numerous pivotal functions of CP within Alphaflexiviridae , including the activation of RNA translation [ 45 ], the facilitation of infection[ 46 ] and the encapsidation of viral genome RNA [ 47 ]. Our findings are in agreement with studies on viral CP from different viral families that documented strong purifying selection pressure along with low genetic diversity [ 43 , 44 , 48 , 49 ]. Altogether, it cannot be ruled out that seashore paspalums may have often been exchanged between countries due to their beneficial use in athletic fields, leading to a low degree of spatial structure in the viral population instead of a very high spatial structure. Furthermore, because of the small number of accessions analyzed, the fact that all accessions were maintained in a single location for multiple years, and the geographic origins could possibly refer to sites where Paspalum plants were maintained before being donated to NPGS [ 50 ], whether the two PaLV populations indeed have different geographic origins and distributions will need to be further confirmed. Further, since the plants have been maintained in controlled environments and in pots it is unlikely that contamination can occur between accessions. Since Paspalum is a vegetatively propagated crop the transmission of the virus and the lower diversity observed is more likely to be preserved long term. Similar observations of low to high genetic differences were also made in PVX populations, where isolates from different continents were analyzed where human-mediated movement influenced viral population dynamics [ 43 , 44 ]. Nevertheless, the diversity observed in PaLV should not impact the efficiency of the RT-PCR detection assay developed in this study. Conclusions In this study, we characterized a novel virus, PaLV, which represents a novel taxon and a tentatively named genus, ‘ Paspalovirus ’, within the family Alphaflexiviridae . While LoLV is the closest recognized virus species to PaLV it is phylogenetically distinct and significant differences in genome organization and the absence of key biological data preclude its classification as a true member of the Lolavirus. PaLV was present in most Paspalum accessions maintained at UGA. Given that seashore paspalum is typically propagated vegetatively through sod, containerized material, stolons, and rhizomes, the virus can be easily disseminated during plant propagation and potentially readily disseminated by mechanical transmission during mowing [ 51 ]. Here, we observed no discernible effect of PaLV infection on Paspalum . However, the asymptomatic nature of PaLV infections does not preclude the possibility of its impact on plant growth, breeding, and other plant physiological factors such as salt tolerance, a key horticultural characteristic, and a subject that requires further investigation. Furthermore, it is plausible that further adoption of Paspalum in the turf industry can potentially result in the emergence of more virulent PaLV strains due to vegetative propagation [52, 5354]. Therefore, PaLV can potentially emerge as a challenge for turfgrass management with the lack of resistance traits in the Paspalum germplasm. Further studies should focus on screening Paspalum germplasm against PaLV through quantitative and semi-quantitative RT-PCR-based methods, similar to what was observed for resistance to fungal diseases in Paspalum [ 7 , 55 ]. Nevertheless, even in the present context, it is essential to implement control methods, such as using virus-free source materials to propagate paspalum. Declarations Ethics approval and consent to participate N/A. Consent for publication N/A. Data availability The datasets generated and analyzed during the current study are available at NCBI. //www.ncbi.nlm.nih.gov/, PV239592 (PaLV isolate Spence), PV239593 (PaLV isolate HI10), PV239594 (PaLV isolate PI509022). Funding We gratefully acknowledge the funding for S.P. and K.D. from NSF award IOS-1915919. Author contributions K.D., J.A.F., B.N.A. conceived and designed the research; S.B., T.F.S., conducted the experiments; A.L.P., T.F.S., A.H-W., S.P., C.D., collected and processed samples; S.B, X.H., P.A. analyzed the data from HTS; S.B., P.A., B.N.A. wrote the initial draft. All authors read, edited, and approved the manuscript Acknowledgements N/A. Competing Interests The author(s) declare no competing interests. References Braun RC, Mandal P, Nwachukwu E, Stanton A. The role of turfgrasses in environmental protection and their benefits to humans: Thirty years later. 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Population genetic analysis of potato virus X based on the CP gene sequence. Virusdisease. 2017;28:93–101. Fuentes S, Gibbs AJ, Hajizadeh M, Perez A, Adams IP, Fribourg CE, et al. The phylogeography of potato virus x shows the fingerprints of its human vector. Viruses. 2021;13. Karpova O V., Arkhipenko M V., Zayakina O V., Nikitin NA, Kiselyova OI, Kozlovsky S V., et al. Regulation of RNA translation in potato virus X RNA-coat protein complexes: The key role of the N-terminal segment of the protein. Mol Biol. 2006;40:628–34. Chapman S, Kavanagh T, Baulcombe D. Potato virus X as a vector for gene expression in plants. The Plant Journal. 1992;2:549–57. Cruz SS, Roberts AG, Prior DAM, Chapman S, Oparka KJ. Cell-to-cell and phloem-mediated transport of Potato virus X: The role of virions. Plant Cell [Internet]. 1998;10:495–510. Available from: https://academic.oup.com/plcell/article/10/4/495/5999278 Bera S, Fraile A, García-Arenal F. Analysis of fitness trade-offs in the host range expansion of an RNA virus, Tobacco mild green mosaic virus . J Virol. 2018;92:1–15. Parizad S, Dizadji A, Koohi Habibi M, Winter S, Kalantari S, Movi S, et al. Description and genetic variation of a distinct species of Potyvirus infecting saffron (Crocus sativus L.) plants in major production regions in Iran. Annals of Applied Biology. 2018; 173(3):233-242.. Eudy D, Bahri BA, Harrison ML, Raymer P, Devos KM. Ploidy level and genetic diversity in the genus Paspalum, group disticha. Crop Sci. 2017;57:3319–32. Shadow RA. SEASHORE PASPALUM Panicum vaginatum Sw. Plant Symbol = PAVA [Internet]. Available from: https://plants.usda.gov/DocumentLibrary/factsheet/pdf/fs_pava.pdf Fraile A, García-Arenal F. Environment and evolution modulate plant virus pathogenesis. Curr Opin Virol. 2016;17:50–6. Hily JM, Poulicard N, Mora MÁ, Pagán I, García-Arenal F. Environment and host genotype determine the outcome of a plant-virus interaction: From antagonism to mutualism. New Phytologist. 2016;209:812–22. Nakamura D, Minato N, Furuya M, Komatsu K, Fuji S ichi. Long-term passages of Plantago asiatica mosaic virus alter virulence and multiplication in Nicotiana benthamiana plants. Arch Virol. 2024;169. Oberti H, Reyno R, Do Canto J, Castro L, Murchio S, Rossi C, et al. An update in Claviceps paspali disease: a comprehensive analysis on field and greenhouse Paspalum spp. Infection . Plant Dis. 2024; Table Table 1: Identity (%) in nucleotide and deduced amino acid (aa) sequences between PaLV and other viruses from Lolavirus, Potexvirus, and Mandarivirus genera. Additional Declarations No competing interests reported. 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Seashore paspalum (\u003cem\u003ePaspalum vaginatum\u003c/em\u003e Sw.\u003cem\u003e)\u003c/em\u003e, a warm-season perennial turfgrass, has attracted attention due to its salinity tolerance and adaptability to coastal environments in tropical and subtropical regions [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. As a result, seashore paspalum is a popular choice for sports fields in coastal regions where freshwater availability is an issue and saltwater irrigation results in tremendous cost savings [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Beyond its functional benefits, seashore paspalum also contributes to environmental sustainability by mitigating the urban heat island effect, providing forage and aesthetic value, and generating economic and social benefits. The species is also reportedly tolerant to flooding and low oxygen conditions [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Due to these numerous benefits, the cultivation area of \u003cem\u003ePaspalum\u003c/em\u003e has expanded beyond coastal areas; however, in new environments, plants are exposed to many unknown pathogens, increasing the risk of disease damage.\u003c/p\u003e \u003cp\u003eSeashore paspalum, similar to other turfgrasses, is propagated through clonal methods, resulting in genetic uniformity [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. As a result, viruses are easily spread across the propagated plant material and introduced into new areas [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Notable viruses that have been identified in \u003cem\u003ePaspalum\u003c/em\u003e, under natural or experimental conditions, include Paspalum striate mosaic virus (PSMV, \u003cem\u003eGeminiviridae\u003c/em\u003e) [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e], sugarcane mosaic virus (SCMV, \u003cem\u003ePotyviridae\u003c/em\u003e) [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e], sugarcane streak R\u0026eacute;union virus (SSRV, \u003cem\u003eGeminiviridae\u003c/em\u003e) [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], Chloris striate mosaic virus (CSMV, \u003cem\u003eGeminiviridae\u003c/em\u003e) [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e], Paspalum dilatatum striate mosaic virus (PDSMV, \u003cem\u003eGeminiviridae\u003c/em\u003e) [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e], and barley yellow dwarf virus (BYDV, \u003cem\u003eTombusviridae\u003c/em\u003e) [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn recent years, high-throughput sequencing (HTS) has emerged as a transformative technology in the field of virology, particularly in the realm of viral discovery and diagnostics [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. This approach has led to the identification of numerous novel viruses in diverse biological samples, including grasses, weeds, trees, and food crops [\u003cspan additionalcitationids=\"CR17 CR18 CR19\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Therefore, in this study we leveraged HTS to investigate the virome of \u003cem\u003ePaspalum\u003c/em\u003e germplasm used in breeding programs. For this purpose, 27 \u003cem\u003eP. vaginatum\u003c/em\u003e and three \u003cem\u003eP. distichum\u003c/em\u003e (a sister species of \u003cem\u003eP. vaginatum\u003c/em\u003e) accessions were obtained from the USDA National Plant Germplasm System (NPGS) collection and maintained at the University of Georgia (UGA), Griffin and Athens campuses.\u003c/p\u003e \u003cp\u003eOur results include the first report of a virus representing a novel taxon within the \u003cem\u003eAlphaflexiviride\u003c/em\u003e that infects both \u003cem\u003eP. vaginatum\u003c/em\u003e and \u003cem\u003eP. distichum\u003c/em\u003e. Confirmation of putative viral sequences was obtained by RT-PCR and genome terminals were completed with 5' RACE. Based on phylogenetic analysis and genomic structure, we propose that this novel virus, tentatively named \u003cem\u003ePaspalovirus paspali\u003c/em\u003e (Paspalum latent virus, PaLV) is a new virus, in the family \u003cem\u003eAlphaflexiviridae\u003c/em\u003e. Here, we report the identification of PaLV, its biological and molecular characterization, phylogenetic relationships, genetic diversity and population structure, and PaLV screening of \u003cem\u003eP. vaginatum\u003c/em\u003e and \u003cem\u003eP. distichum\u003c/em\u003e germplasm maintained in greenhouses at UGA.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e"},{"header":"Material and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePlant Material and Growth\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003e \u003cem\u003ePaspalum\u003c/em\u003e plants, obtained initially from the USDA-NPGS collection, Griffin, were maintained in greenhouses at the UGA Griffin and Athens campuses in 4x4 pots. The soil was a 1:1 mixture of sand and Miracle-Gro potting mix. Plants were fertilized once every two weeks with Osmocote Plus.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eTotal Plant RNA Extraction\u003c/h3\u003e\n\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eTotal RNA from the \u003cem\u003ePaspalum\u003c/em\u003e accession PI 509022 leaf tissue was extracted using a RNeasy Plant Mini Kit (Qiagen, Hilden, Germany), and an RNase-Free DNase Set (Qiagen) was used for DNA digestion. The manufacturer\u0026rsquo;s recommended protocol was used without modifications. For accessions HI10 and Spence, papillae were peeled from the adaxial surface of leaves (second fully open, developed leaf on growing stolon segment) from plants grown under freshwater (0 mM NaCl) and salt stress (200 mM NaCl) irrigation for six weeks. Strings of papillae were collected in 2 mL tubes resting on a liquid nitrogen bath to maintain RNA integrity, and frozen tissues were ground using a TissueLyser II bead mill (Qiagen). The ground tissue was homogenized in 1 mL of TRIzol reagent (Invitrogen, Waltham, MA) per 100 mg of tissue, followed by phase separation after adding 200 \u0026micro;L of chloroform. The RNA-containing aqueous phase was carefully transferred to a 2 mL nuclease-free tube and mixed with an equal volume of 100% ethanol. The resulting RNA-ethanol mixture was loaded onto a Zymo-Spin IC column from the Zymo RNA Clean \u0026amp; Concentrator-5 kit (Zymo Research, Irvine, CA), and RNA clean-up was performed following the manufacturer\u0026rsquo;s instructions. RNA was eluted in 10 \u0026micro;L nuclease-free water and RNA concentration was determined using a NanoDrop spectrophotometer, and the quality was assessed via electrophoresis on a 1% Tris-Borate-EDTA (TBE) agarose gel.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e\n\u003ch3\u003eHTS and Virus Identification\u003c/h3\u003e\n\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eThe RNA extracts from leaves of accession PI 509022 at USDA APHIS-PPQ, Beltsville, and from adaxial leaf papillae from accessions HI 10 and Spence at the University of Georgia, Athens, were subjected to HTS at their respective place of extraction. For HTS of RNA from PI 509022, a single-indexed ribosomal-RNA-depleted cDNA library was prepared using the TruSeq\u0026reg; Stranded Total RNA Library Prep Plant kit (Illumina, San Diego, CA) and sequenced on an Illumina NextSeq 500 platform, generating 19,815,890 single-end, 75-bp reads. For HTS of RNA from accessions HI 10 and Spence, barcoded stranded mRNA-seq libraries were generated from 500 ng of RNA with the KAPA Stranded mRNA-Seq Kit (Roche, Basel, Switzerland) according to the manufacturer's instructions, using half-reactions for all steps except the final library amplification. The concentration of the libraries was measured using a Qubit Fluorometer with the Qubit 1X dsDNA High Sensitivity Assay Kit (Invitrogen). Subsequently, 30 ng of each library was pooled into a larger set of 24 leaf and peels libraries and loaded onto a single flow cell of an Illumina NextSeq 2000 for paired end (PE) 150 bp sequencing at the Georgia Genomics and Bioinformatics Core (GGBC) at UGA. Reads from HTS were assembled using the de novo assembly tool in PhytoPipe (2023) [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e] and compared with viral pathogen databases. Briefly, raw reads were filtered and trimmed using Trimmomatic (v.0.39) [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Then, contigs were assembled using Trinity (v2.8.6) [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] and compared to the NCBI Viral Reference Database (Jan 2022) by BLASTn search [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] and to the Reference Viral Database protein database (v22)[\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e] by Diamond blastx search [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Default parameters were used in all programs.\u003c/p\u003e \u003cp\u003e \u003cb\u003e5' RACE\u003c/b\u003e \u003c/p\u003e \u003cp\u003eRapid amplification of cDNA ends with the SMARTer\u0026reg; RACE 5\u0026prime;/3\u0026prime; Kit (Takara) was used to complete the 5\u0026prime; end of the ssRNA segment with the RACE primer \u0026ndash; 5'\u003cb\u003e-\u003c/b\u003eGGACCACTGTGGCGTAGAGGATGTCGAGGT-3'; the 5' end of the primer was linked to the adapter sequence provided by the company. All PCR fragments were sequenced by direct Sanger sequencing in both directions, and sequences were aligned to the reference HTS-derived contigs using Geneious (v11.0.3).\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e\n\u003ch3\u003eRT-PCR Validation of HTS and Screening of Germplasm\u003c/h3\u003e\n\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eA one-step RT-PCR protocol was developed for detection of PaLV using the SuperScript\u0026trade; III One-Step RT-PCR System with Platinum\u0026reg; Taq DNA Polymerase (Invitrogen). Primer pairs ORF1.FP: 5\u0026prime;-AAACAAGTGGAATTTCACGGG-3\u0026prime; and ORF1.RP: 5\u0026prime;-CTGCTCTGGTGAGAAGATATCG-3\u0026prime;, were designed to amplify a 605 bp amplicon from the replicase gene. Cycling parameters were used following the manufacturer's guidelines except for the 62\u0026deg;C annealing temperature.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e\n\u003ch3\u003eHost Range Studies\u003c/h3\u003e\n\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eAfter confirmation of PaLV infection in \u003cem\u003ePaspalum\u003c/em\u003e through RT-PCR, infected plant tissues were freshly ground in 0.01M potassium phosphate buffer (pH 7) for mechanical inoculation. All plants were sprayed with carborundum, an abrasive, before rub-inoculation. Mechanical inoculation of sap was conducted on nine different plant species: \u003cem\u003eHordeum vulgare, Triticum aestivum, Zea mays, Sorghum spp., Dactylis glomerata, Setaria italica, Lolium multiflorum, Miscanthus sacchariflorus\u003c/em\u003e, and \u003cem\u003eAvena sativa\u003c/em\u003e. Three biological replicates were used for each species. All bioassay plants were kept in controlled-temperature greenhouse conditions at 26℃ \u0026plusmn; 2℃ with 16 h light and 8 h dark. Two plant species, \u003cem\u003eH. vulgare\u003c/em\u003e, and \u003cem\u003eT. aestivum\u003c/em\u003e were assayed in a growth chamber maintained at 16℃ with 16 h light and 8 h dark.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eSequence, Phylogenetic, and Recombination Analysis\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eThe viral sequences were analyzed by BLASTn (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://blast.ncbi.nlm.nih.gov/Blast.cgi\u003c/span\u003e\u003cspan address=\"https://blast.ncbi.nlm.nih.gov/Blast.cgi\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) with default parameters. The putative proteins and potential open reading frames (ORFs) were determined using ORFfinder (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ncbi.nlm.nih.gov/orffinder/\u003c/span\u003e\u003cspan address=\"https://www.ncbi.nlm.nih.gov/orffinder/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) and subsequent blastp annotation by using the non-redundant protein sequences database. Pairwise comparisons between related viral ORFs were performed with Sequence Demarcation Tool (SDT) v1.2 to calculate the identity percentage for each ORF [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Conserved domains within these proteins were identified using the Conserved Domain Database (CDD) [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. The coat protein (CP) sequences were modeled using AlphaFold2[\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. PaLV CP (CP\u003csub\u003ePaLV\u003c/sub\u003e) was superimposed on the Lolium latent virus (CP\u003csub\u003eLoLV\u003c/sub\u003e) using TM-align [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Structural similarity between CP\u003csub\u003ePaLV\u003c/sub\u003e and CP\u003csub\u003eLoLV\u003c/sub\u003e was further evaluated based on TM scores, using 0.5 as a cut-off. Superimposed images were exported from TM-align and loaded into Chimera X [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e] for further processing.\u003c/p\u003e \u003cp\u003eEvolutionary relationship of PaLV with representative species from the genera, \u003cem\u003eLolavirus\u003c/em\u003e, \u003cem\u003ePotexvirus\u003c/em\u003e, \u003cem\u003eMandarivirus\u003c/em\u003e, and \u003cem\u003eAllexivirus\u003c/em\u003e in the \u003cem\u003eAlphaflexiviridae\u003c/em\u003e family were included in a maximum-likelihood phylogenetic tree based on amino acid sequences of replicase using the LG with gamma distribution and frequency substitution model previously inferred using jModelTest in MEGA 11 [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. The phylogenic analysis was performed with 1,000 bootstrap replicates and potato virus T as an outgroup. The resulting phylogenetic tree was visualized and formatted using FigTree v1.4.4 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://tree.bio.ed.ac.uk/software/figtree/\u003c/span\u003e\u003cspan address=\"http://tree.bio.ed.ac.uk/software/figtree/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe Recombination Detection Program V.5 (RDP5) program was used to explore the occurrence of recombination events in full-length viral genome sequences. Various methods implemented in RDP5 [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e] including RDP, SisterScan, Bootscan, Chimaera, GeneConv, MaxChi, and 3Seq algorithms, were used. Any recombinant events with \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001 predicted by four of the seven prediction methods were considered reliable and used in the analysis.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eGenetic diversity\u003c/h3\u003e\n\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eThe genetic diversity of the PaLV population was analyzed using 11 PaLV isolates obtained from different \u003cem\u003ePaspalum\u003c/em\u003e accessions from the UGA collection. Genetic diversity was calculated based on the complete CP gene nucleotide sequences. The primer set designed to amplify the CP region of PaLV was developed according to the consensus sequence of three PaLV isolates obtained through HTS (CP_F: 5\u0026prime;-GATCGGAAGCCTCAGTTGTG-3\u0026prime;; CP_R: 5\u0026prime;-CGGTGGCCAGGGTAGATTAA-3\u0026prime;). The CP nucleotide sequence of 11 PaLV isolates was aligned using the Muscle program implemented in MEGA 11 and the mean genetic diversity for the entire population was calculated. The average number of non-synonymous (d\u003csub\u003eN\u003c/sub\u003e) and synonymous (d\u003csub\u003eS\u003c/sub\u003e) nucleotide substitutions per site and the dN/dS ratio as an estimate of the CP's selection pressure were computed using the DnaSP v.5 software [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. The Wright\u0026rsquo;s fixation index statistic \u003cem\u003eF\u003c/em\u003e\u003csub\u003e\u003cem\u003eST\u003c/em\u003e\u003c/sub\u003e[\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e] was calculated using DnaSP v.5. Frequent gene flow is considered to occur when \u003cem\u003eF\u003c/em\u003e\u003csub\u003e\u003cem\u003eST\u003c/em\u003e\u003c/sub\u003e \u0026lt; 0.33.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eGenome Structure of PaLV\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eHigh-throughput sequencing and 5\u0026prime; RACE revealed the complete genome of PaLV to be 6,995 nucleotides (nt) in length, excluding the poly(A) tail. The genome comprised five open reading frames (ORFs), flanked by an 81 nt 5\u0026prime; untranslated region (UTR) and an 86 nt 3\u0026prime; UTR. Pairwise comparisons of PaLV ORFs with representative \u003cem\u003eAlphaflexiviridae\u003c/em\u003e viruses showed highest identity with Lolium latent virus (LoLV), ranging from 49\u0026ndash;57% at the nucleotide level and 26\u0026ndash;55% at the amino acid level (Table\u0026nbsp;1).\u003c/p\u003e \u003cp\u003eORF1 encoded a 173-kDa replicase protein containing conserved domains: Methyltransferase (aa 39\u0026ndash;329), Viral RNA helicase (aa 768\u0026ndash;997), and RNA-dependent RNA polymerase (aa 1178\u0026ndash;1474) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). The AlkB domain, which is present in LoLV, was absent in ORF 1 of PaLV. ORFs 2\u0026ndash;4 encoded triple gene block (TGB) proteins: TGB1 (30.4 kDa), TGB2 (13.5 kDa), and TGB3 (7.8 kDa). These proteins have been shown to play a crucial role in virus movement [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. A putative octanucleotide promoter sequence for sgRNA1 (CUUAAGUU) was identified 21 nt upstream of ORF2, overlapping the stop codon of ORF1 (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003eORF5 encoded the coat protein (CP) of 35.4 kDa with a start codon in-frame and immediately followed the stop codon of ORF4, with no intergenic region (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003eC). A putative sgRNA2 promoter sequence (GUUAAGUU) was located 14 nt upstream of ORF5, within ORF4. ORF6, present in LoLV and other tentative lolaviruses such as Weeping alkaligrass lolavirus 1 (WaLoV1), that overlaps the 3\u0026prime; end of ORF 5, was not identified in PaLV or the other tentative lola-like members, such as Foxtail millet lolavirus 1 (FmLoV1) [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003eA).\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eComparison of CP\u003csub\u003ePaLV\u003c/sub\u003e with CP\u003csub\u003eLoLV\u003c/sub\u003e\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eSubcellular localization prediction using Plant-mSubP indicated that CP\u003csub\u003ePaLV\u003c/sub\u003e was likely localized in cytoplasm or peroxisomes, while CP\u003csub\u003eLoLV\u003c/sub\u003e and CP\u003csub\u003ePVX\u003c/sub\u003e were predicted to localize in plastids. Subsequently, the protein structure of CP\u003csub\u003ePaLV\u003c/sub\u003e and CP\u003csub\u003eLoLV\u003c/sub\u003e was modeled to compare their structural relatedness. For this purpose, both amino acid sequences were subjected to Alphafold2 protein structure predictions (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The structure of CP\u003csub\u003ePaLV\u003c/sub\u003e was predicted with a lower confidence level than that of CP\u003csub\u003eLoLV\u003c/sub\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The structural modeling of both CPs revealed the presence of three domains: i) flexible N-terminal, ii) the core, and iii) C-terminal extensions. The N-terminus of PaLV does not contain a proteolytic cleavage site for chloroplast transit peptide that yields a smaller CP isoform in LoLV. However, a smaller ORF of 28kDa was predicted within the CP ORF. The core and C-terminal regions of the CP play a crucial role in protein-protein and protein-RNA interactions and have been found to show the highest degree of structural conservation in \u003cem\u003eAlphaflexiviridae\u003c/em\u003e [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. Consequently, a comparative analysis of the core and C-terminal domains (151\u0026ndash;300 and 106\u0026ndash;280 aa in CP\u003csub\u003ePaLV\u003c/sub\u003e and CP\u003csub\u003eLoLV\u003c/sub\u003e, respectively) of both CPs was conducted. As predicted, a high TM score, indicative of high structural similarity, was obtained upon the superimposition of the core and C-terminal regions of both CPs, which consists of conserved α-helical secondary structures (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003ePhylogenetic Analysis\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eMaximum-likelihood phylogenetic analysis based on replicase amino acid sequences placed PaLV in a distinct clade clearly separated from the \u003cem\u003ePotexvirus\u003c/em\u003e and \u003cem\u003eLolavirus\u003c/em\u003e genera. The analysis also included putative viral sequences obtained through data mining and submitted as Third-Party Annotation (TPA), such as Saltwater paspalum lolavirus 1 (SpLoV1, synonym PaLV [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e], FmLoV1, a Lola-like virus from Maize (Lola maydis), and WaLoV1. Their inclusion further supports the formation of this novel group within the \u003cem\u003eAlphaflexiviridae\u003c/em\u003e family.\u003c/p\u003e \u003cp\u003eAll PaLV isolates along with SpLoV1formed a unique and well-supported clade (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Notably, SpLoV1 shared 97% nucleotide identity with PaLV, which indicates the same viral species. FmLoV1 and Lola maydis sequences represent two potential new members within the PaLV clade. Whereas WaLoV1 appeared to cluster within the \u003cem\u003eLolavirus\u003c/em\u003e clade. No recombination events were detected in PaLV genomes using any of the algorithms implemented in RDP5.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eGenetic Diversity of PaLV Population\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eAnalysis of eleven CP sequences from PaLV isolates revealed two distinct clusters: Cluster A (primarily U.S. origin) and Cluster B (primarily rest of world). F\u003csub\u003eST\u003c/sub\u003e value was 0.124, indicating low spatial genetic structure. To ascertain whether the low diversity could be attributed to selection, the dN/dS ratio was estimated, yielding a value of 0.042, suggesting the presence of strong purifying (negative) selection on CP. This finding is further corroborated by the observation of low genetic diversity (0.031\u0026thinsp;\u0026plusmn;\u0026thinsp;0.004) of CP.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eHost Range Studies\u003c/h2\u003e \u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eAll inoculated (virus and mock) plants were examined by RT-PCR after nine indicator plants representing nine plant genera were subjected to mechanical inoculation with leaf extract from PaLV-infected plants. All inoculated plants of the nine assayed genera were monitored over a period of 21 days post-inoculation (dpi) for symptom development. (Supp Table\u0026nbsp;1). PaLV amplicons of the expected size were obtained with RT-PCR in: \u003cem\u003eZea mays\u003c/em\u003e, \u003cem\u003eSorghum\u003c/em\u003e spp., \u003cem\u003eSetaria italica\u003c/em\u003e, and \u003cem\u003eLolium multiflorum\u003c/em\u003e. Interestingly, \u003cem\u003eT. aestivum\u003c/em\u003e plants maintained at 16℃ were also susceptible to PaLV infection based on RT-PCR. The infection of the aforementioned species indicates they are hosts of PaLV. On the contrary, no amplicons were amplified from the remaining inoculated species: \u003cem\u003eHordeum vulgare\u003c/em\u003e, \u003cem\u003eAvena sativa\u003c/em\u003e, \u003cem\u003eDactylis glomerata\u003c/em\u003e, and \u003cem\u003eMiscanthus sacchariflorus\u003c/em\u003e. None of the experimental hosts showed any symptoms of virus infection. Similarly, no symptoms were also observed in infected seashore paspalum germplasm. No symptoms were observed in mock-inoculated plants. In contrast, LoLV infections in \u003cem\u003eLolium\u003c/em\u003e plants were reported either to be asymptomatic or showing mild chlorotic streaking on the leaves[\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e] and in \u003cem\u003eNicotiana benthamiana\u003c/em\u003e plants systemic symptoms such as mosaic and vein netting showed at 15 dpi [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eScreening of Paspalum Germplasm\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eA one-step RT-PCR assay targeting ORF1 was developed. A primer pair was designed to target the RNA-dependent RNA polymerase domain in ORF1, amplifying a region of 605 bp. Thirty different \u003cem\u003ePaspalum\u003c/em\u003e accessions maintained at UGA were analyzed. Two samples of butterfly bush were used as a negative control, and a PaLV-positive plant as a positive control. The RT-PCR results revealed the presence of PaLV infection in 27 out of 30 accessions (Supp Table\u0026nbsp;2), with an incidence of 90% in the tested \u003cem\u003ePaspalum\u003c/em\u003e accessions. Notably, none of the negative controls showed the presence of PaLV, indicating the specificity of the primer pair.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eIn this study, we report the molecular and biological characterization of \u003cem\u003ePaspalum latent virus\u003c/em\u003e (PaLV), a novel virus infecting seashore paspalum and representing a new taxon within the family \u003cem\u003eAlphaflexiviridae\u003c/em\u003e. The complete genome structure, phylogenetic placement, host range, and population diversity of PaLV were investigated, revealing several unique features that distinguish it from known members of the \u003cem\u003eLolavirus\u003c/em\u003e genus. Although PaLV shares moderate sequence identity with \u003cem\u003eLolium latent virus\u003c/em\u003e (LoLV), its genome organization diverges significantly. The absence of the AlkB domain in the replicase, absence of a proteolytic cleavage site within the CP, and the overlapping start-stop codon arrangement between ORF4 and ORF5 suggest distinct evolutionary adaptations. This is a notable divergence from LoLV. Sister members of the PaLV clade, FmLoV1 and Lola maydis, also lack the AlkB domain within the replicase. On the other hand, WaLoV1, which clusters with LoLV contains the AlkB domain. The AlkB domain is speculated to play a crucial role in maintaining the stability of RNA genomes, which are susceptible to methylation due to pesticide applications [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. This speculation led to the hypothesis that the acquisition of the AlkB domain is a relatively recent development in the evolution of viruses within the \u003cem\u003eAlphaflexiviridae\u003c/em\u003e family [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. Furthermore, PaLV lacks another signature feature of lolaviruses, ORF6, which is present also in WaLoV1 PaLV shows similar protein sizes for the various ORFs to FmLoV1 rather than LoLV and WaLoV1[\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. Overall, the absence and presence of these signature features across both clades further support the divergence across both taxa. As a result, this supports our hypothesis that PaLV represents a new genus, tentatively named \u0026lsquo;\u003cem\u003ePaspalovirus\u003c/em\u003e\u0026rsquo;.\u003c/p\u003e \u003cp\u003eBased on the ICTV demarcation criteria in the family \u003cem\u003eAlphaflexiviridae\u003c/em\u003e, viruses are separated into different genera when their coat protein or polymerase genes share less than about 45% amino acid identity. Whereas, viruses are considered distinct species if they share less than 72% nucleotide identity or less than 80% amino acid identity in either their coat protein (CP) or replicase genes. In this study, PaLV replicase showed less than 45% amino acid identity with LoLV, which is below the genus demarcation threshold and the CP amino acid identity was below 40%. Further, there is incongruency in other features, such as different genome organization and putative protein sizes, in placing PaLV within the \u003cem\u003eLolavirus\u003c/em\u003e or other established genera.\u003c/p\u003e \u003cp\u003eDespite low amino acid identity between PaLV and LoLV CPs, structural modeling revealed some conserved core and C-terminal domains. These domains are analogous to the crystal structures of CPs of PapMV and \u003cem\u003ePepino mosaic virus\u003c/em\u003e from the \u003cem\u003ePotexvirus\u003c/em\u003e genus [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e, \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. The N- and C-terminal extensions play a crucial role in the polymerization of CP [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. The core and C-terminal regions of the CP play a crucial role in protein-protein and protein-RNA interactions and have been found to show the highest degree of structural conservation in \u003cem\u003eAlphaflexiviridae\u003c/em\u003e [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. These regions are critical for virion assembly and RNA binding, suggesting functional conservation. Interestingly, subcellular localization predictions indicated that PaLV CP may localize to cytoplasm or peroxisomes, unlike LoLV and PVX CPs, which preferentially target plastids. This divergence coupled with a very different N-terminus sequence within PaLV CP compared to LoLV may influence host interactions and symptom expression. For instance, tentative members FmLoV1 and Lola maydis have several internal smaller ORF within the CP. Further experimentation beyond the scope of this work is needed to validate whether one or more CP isoforms exist for paspaloviruses.\u003c/p\u003e \u003cp\u003ePaLV was detected in multiple grass species, including \u003cem\u003eZea mays\u003c/em\u003e, \u003cem\u003eSorghum spp.\u003c/em\u003e, \u003cem\u003eSetaria italica\u003c/em\u003e, and \u003cem\u003eLolium multiflorum\u003c/em\u003e, indicating a broad host range. However, no visible symptoms were observed in any infected plants, including paspalum accessions. This latent infection profile complicates field diagnostics and may contribute to unnoticed virus spread through vegetative propagation. In contrast, LoLV infections in \u003cem\u003eLolium\u003c/em\u003e plants were reported either to be asymptomatic or showing mild chlorotic streaking on the leaves [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e] and in experimental plants systemic symptoms such as mosaic and vein netting showed at 15 dpi [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. This indicates that while PaLV shares similarity to LoLV, PaLV does not show severe or adverse effects due to the lack of virulence factors which can allow the virus to go undetected in some grasses.\u003c/p\u003e \u003cp\u003eAnalysis of 11 PaLV isolates revealed low genetic diversity and strong purifying selection on the CP gene. A \u003cem\u003eF\u003c/em\u003e\u003csub\u003e\u003cem\u003eST\u003c/em\u003e\u003c/sub\u003e of 0.124 indicated a low degree of spatial structure and genetic differences between the population [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. These results are consistent with the expectation of CP sequence conservation given the numerous pivotal functions of CP within \u003cem\u003eAlphaflexiviridae\u003c/em\u003e, including the activation of RNA translation [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e], the facilitation of infection[\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e] and the encapsidation of viral genome RNA [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. Our findings are in agreement with studies on viral CP from different viral families that documented strong purifying selection pressure along with low genetic diversity [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. Altogether, it cannot be ruled out that seashore paspalums may have often been exchanged between countries due to their beneficial use in athletic fields, leading to a low degree of spatial structure in the viral population instead of a very high spatial structure.\u003c/p\u003e \u003cp\u003eFurthermore, because of the small number of accessions analyzed, the fact that all accessions were maintained in a single location for multiple years, and the geographic origins could possibly refer to sites where \u003cem\u003ePaspalum\u003c/em\u003e plants were maintained before being donated to NPGS [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e], whether the two PaLV populations indeed have different geographic origins and distributions will need to be further confirmed. Further, since the plants have been maintained in controlled environments and in pots it is unlikely that contamination can occur between accessions. Since Paspalum is a vegetatively propagated crop the transmission of the virus and the lower diversity observed is more likely to be preserved long term. Similar observations of low to high genetic differences were also made in PVX populations, where isolates from different continents were analyzed where human-mediated movement influenced viral population dynamics [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. Nevertheless, the diversity observed in PaLV should not impact the efficiency of the RT-PCR detection assay developed in this study.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eIn this study, we characterized a novel virus, PaLV, which represents a novel taxon and a tentatively named genus, \u0026lsquo;\u003cem\u003ePaspalovirus\u003c/em\u003e\u0026rsquo;, within the family \u003cem\u003eAlphaflexiviridae\u003c/em\u003e. While LoLV is the closest recognized virus species to PaLV it is phylogenetically distinct and significant differences in genome organization and the absence of key biological data preclude its classification as a true member of the Lolavirus. PaLV was present in most \u003cem\u003ePaspalum\u003c/em\u003e accessions maintained at UGA. Given that seashore paspalum is typically propagated vegetatively through sod, containerized material, stolons, and rhizomes, the virus can be easily disseminated during plant propagation and potentially readily disseminated by mechanical transmission during mowing [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. Here, we observed no discernible effect of PaLV infection on \u003cem\u003ePaspalum\u003c/em\u003e. However, the asymptomatic nature of PaLV infections does not preclude the possibility of its impact on plant growth, breeding, and other plant physiological factors such as salt tolerance, a key horticultural characteristic, and a subject that requires further investigation. Furthermore, it is plausible that further adoption of \u003cem\u003ePaspalum\u003c/em\u003e in the turf industry can potentially result in the emergence of more virulent PaLV strains due to vegetative propagation [52, 5354]. Therefore, PaLV can potentially emerge as a challenge for turfgrass management with the lack of resistance traits in the \u003cem\u003ePaspalum\u003c/em\u003e germplasm. Further studies should focus on screening \u003cem\u003ePaspalum\u003c/em\u003e germplasm against PaLV through quantitative and semi-quantitative RT-PCR-based methods, similar to what was observed for resistance to fungal diseases in \u003cem\u003ePaspalum\u003c/em\u003e [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e]. Nevertheless, even in the present context, it is essential to implement control methods, such as using virus-free source materials to propagate paspalum.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eN/A.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eN/A.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp;The datasets generated and analyzed during the current study are available at NCBI. //www.ncbi.nlm.nih.gov/, PV239592 (PaLV isolate Spence), PV239593 (PaLV isolate HI10), PV239594 (PaLV isolate PI509022).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe gratefully acknowledge the funding for S.P. and K.D. from NSF award IOS-1915919.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eK.D., J.A.F., B.N.A. conceived and designed the research; S.B., T.F.S., conducted the experiments; A.L.P., T.F.S., A.H-W., S.P., C.D., collected and processed samples; S.B, X.H., P.A. analyzed the data from HTS; S.B., P.A., B.N.A. wrote the initial draft. All authors read, edited, and approved the manuscript\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eN/A.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe author(s) declare no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eBraun RC, Mandal P, Nwachukwu E, Stanton A. The role of turfgrasses in environmental protection and their benefits to humans: Thirty years later. Crop Sci [Internet]. 2024;64:2909\u0026ndash;44. Available from: https://acsess.onlinelibrary.wiley.com/doi/10.1002/csc2.21383\u003c/li\u003e\n\u003cli\u003eWu P, Cogill S, Qiu Y, Li Z, Zhou M, Hu Q, et al. Comparative transcriptome profiling provides insights into plant salt tolerance in seashore paspalum (Paspalum vaginatum). BMC Genomics. 2020;21:1\u0026ndash;16. \u003c/li\u003e\n\u003cli\u003eSpiekerman JJ, Devos KM. 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Curr Opin Virol. 2016;17:50\u0026ndash;6. \u003c/li\u003e\n\u003cli\u003eHily JM, Poulicard N, Mora M\u0026Aacute;, Pag\u0026aacute;n I, Garc\u0026iacute;a-Arenal F. Environment and host genotype determine the outcome of a plant-virus interaction: From antagonism to mutualism. New Phytologist. 2016;209:812\u0026ndash;22. \u003c/li\u003e\n\u003cli\u003eNakamura D, Minato N, Furuya M, Komatsu K, Fuji S ichi. Long-term passages of Plantago asiatica mosaic virus alter virulence and multiplication in Nicotiana benthamiana plants. Arch Virol. 2024;169. \u003c/li\u003e\n\u003cli\u003eOberti H, Reyno R, Do Canto J, Castro L, Murchio S, Rossi C, et al. An update in Claviceps paspali disease: a comprehensive analysis on field and greenhouse Paspalum spp. Infection . Plant Dis. 2024; \u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Table","content":"\u003cp\u003e\u003cstrong\u003eTable 1: Identity (%) in nucleotide and deduced amino acid (aa) sequences between PaLV and other viruses from \u003cem\u003eLolavirus,\u003c/em\u003e \u003cem\u003ePotexvirus,\u0026nbsp;\u003c/em\u003eand\u003cem\u003e\u0026nbsp;Mandarivirus\u003c/em\u003e genera.\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cimg 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Despite its economic importance, viral infections in paspalum remain poorly characterized. This study reports the discovery and characterization of a novel virus infecting paspalum, tentatively named Paspalum latent virus (PaLV), representing a new taxon within the family \u003cem\u003eAlphaflexiviridae\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods \u003c/strong\u003eHigh-throughput sequencing, RACE PCR, and Sanger sequencing were used to obtain complete genome sequences of PaLV from multiple paspalum accessions. Phylogenetic analysis was performed using replicase protein sequences. Structural modeling of the coat protein (CP) was conducted using AlphaFold2 and TM-align. RT-PCR assays were developed for virus detection and host range studies. Genetic diversity and population structure were assessed using CP sequences from 11 PaLV isolates.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults \u003c/strong\u003ePaLV possesses a 6,995 nt long genome with five ORFs and lacks the AlkB domain in its polymerase gene . The genome exhibits unique features, including overlapping start-stop codons between ORF4 and ORF5 and a second in-frame AUG codon in the CP region. Phylogenetic analysis placed PaLV in a distinct clade clearly separated from \u003cem\u003ePotexvirus \u003c/em\u003eand\u003cem\u003e Lolavirus\u003c/em\u003e. Structural modeling revealed conserved CP domains despite low sequence identity. Genetic analysis indicated low diversity and strong purifying selection, with limited spatial structure. An RT-PCR detection assay was developed to detect PaLV in paspalum accessions which was detected at an incidence rate of 90% . Mechanical transmission resulted in infection of \u003cem\u003eZea mays\u003c/em\u003e, \u003cem\u003eSorghum spp.\u003c/em\u003e, \u003cem\u003eSetaria italica\u003c/em\u003e, and \u003cem\u003eLolium multiflorum\u003c/em\u003e, without visible symptoms.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions \u003c/strong\u003ePaLV represents a novel viral species and likely a new tentatively named genus, \u003cem\u003ePaspalovirus\u003c/em\u003e, within the family \u003cem\u003eAlphaflexiviridae\u003c/em\u003e. Its latent infection profile and broad host range underscore the need for routine molecular diagnostics in turfgrass management. The RT-PCR assay developed here provides a reliable tool for PaLV detection, and further studies are warranted to assess its impact on plant physiology and breeding.\u003c/p\u003e","manuscriptTitle":"Molecular and biological characterization of a newly identified virus representing a novel taxon of Alphaflexiviridae infecting different accessions of seashore paspalum, a turfgrass, widely grown in the United States","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-05 10:33:19","doi":"10.21203/rs.3.rs-8745768/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"caf1f0a5-eaf4-4a63-ba0b-b62b7ce63281","owner":[],"postedDate":"February 5th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-04-15T07:56:48+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-05 10:33:19","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8745768","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8745768","identity":"rs-8745768","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}