Pea Enation Mosaic Virus Research Articles

Viral condensates formed by Pea enation mosaic virus 2 sequester ribosomal components and suppress translation

Viral proteins with intrinsic disorder, such as the p26 movement protein from Pea enation mosaic virus 2 (PEMV2), can phase separate and form condensates that aid specific stages of virus replication. However, little is known about the impact of viral condensate formation on essential cellular processes, like translation. In this study, we performed mass spectrometry on affinity-purified p26 condensates and found an enrichment of RNA-binding proteins involved in translation and ribosome biogenesis. Puromycin assays and polysome profiling show that ectopic p26 expression suppresses ribosome assembly and translation in Nicotiana benthamiana, mirroring defects in late-stage PEMV2 infection. Despite interactions with the 2'-O-methyltransferase fibrillarin, p26 does not inhibit translation by altering rRNA methylation but instead binds directly to rRNAs and decreases their solubility. Disruption of ribosome assembly and translation by p26 during late PEMV2 infection may promote stages of the virus replication cycle that are incompatible with translation, including systemic movement.

Effects of Soil Rhizobia Abundance on Interactions between a Vector, Pathogen, and Legume Plant Host.

Soil rhizobia promote nitrogen fixation in legume hosts, maximizing their tolerance to different biotic stressors, plant biomass, crop growth, and yield. While the presence of soil rhizobia is considered beneficial for plants, few studies have assessed whether variation in rhizobia abundance affects the tolerance of legumes to stressors. To address this, we assessed the effects of variable soil rhizobia inoculum concentrations on interactions between a legume host (Pisum sativum), a vector insect (Acyrthosiphon pisum), and a virus (Pea enation mosaic virus, PEMV). We showed that increased rhizobia abundance reduces the inhibitory effects of PEMV on the nodule formation and root growth in 2-week-old plants. However, these trends were reversed in 4-week-old plants. Rhizobia abundance did not affect shoot growth or virus prevalence in 2- or 4-week-old plants. Our results show that rhizobia abundance may indirectly affect legume tolerance to a virus, but effects varied based on plant age. To assess the mechanisms that mediated interactions between rhizobia, plants, aphids, and PEMV, we measured the relative expression of gene transcripts related to plant defense signaling. Rhizobia concentrations did not strongly affect the expression of defense genes associated with phytohormone signaling. Our study shows that an abundance of soil rhizobia may impact a plant's ability to tolerate stressors such as vector-borne pathogens, as well as aid in developing sustainable pest and pathogen management systems for legume crops. More broadly, understanding how variable rhizobia concentrations can optimize legume-rhizobia symbiosis may enhance the productivity of legume crops.

Comparison of Sequencing Methods for Obtaining a Whole Genome of Pea Necrotic Yellow Dwarf Virus from U.K. Peas

During a project investigating pea viruses in the United Kingdom and developing a workflow for high-throughput sequencing in crop surveillance, pea necrotic yellow dwarf virus (PNYDV) was identified at one site, along with pea enation mosaic virus-1, pea enation mosaic virus-2, pea enation mosaic virus satellite RNA, pea seedborne mosaic virus, and turnip yellows virus. Following an initial finding of small PNYDV contigs by an RNA sequencing protocol on MiSeq (Illumina), confirmation and prevalence testing by PCR and Sanger sequencing determined the average PNYDV prevalence to be 3%. This is the first report of PNYDV, and a nanovirus, in the United Kingdom. Initial characterization of the octopartite genome by PCR revealed that component DNA N had a truncated sequence that differed from those previously reported. This led to confirmation testing using rolling circle amplification (RCA) into MiSeq and then MinION (Oxford Nanopore). Sequences were obtained by both RCA-MiSeq and RCA-MinION, with MinION producing more complete genomes than RCA-MiSeq. Data obtained from sequencing were used to provide an initial estimation of the genomic formula of PNYDV. Previous work has looked into comparing each method; however, this study shows how these methods can be used together. The truncated sequence for DNA N was also found by RCA-MiSeq and RCA-MinION and confirmed by specific primers, but further work is required to understand the impact of this truncation. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Endosymbionts modulate virus effects on aphid-plant interactions

Vector-borne pathogens frequently modify traits of their primary hosts and vectors in ways that influence disease transmission. Such effects can themselves be altered by the presence of other microbial symbionts, yet we currently have limited understanding of these interactions. Here we show that effects of pea enation mosaic virus (PEMV) on interactions between host plants and aphid vectors are modulated by the presence of different aphid endosymbionts. In a series of laboratory assays, we found strong interactive effects of virus infection and endosymbionts on aphid metabolomic profiles, population growth, behavior, and virus transmission during aphid feeding. Furthermore, the strongest effects—and those predicted to favor virus transmission—were most apparent in aphid lines harboring particular endosymbionts. These findings show that virus effects on host-vector interactions can be strongly influenced by other microbial symbionts and suggest a potentially important role for such interactions in disease ecology and evolution.

Experimental evidence reveals that vector host preference and performance across host plants is not altered by vector-borne plant viruses

Many plant pathogens are transmitted by generalist insects that move between host species, and vector preference and performance on different hosts affects pathogen transmission. Most studies of host–vector–pathogen interactions focus on a single host, however, and it is relatively unknown if viruses affect the host breadth of vectors. For example, although pea aphids (Acyrthosiphon pisum) often form tight associations with specific hosts, viruses transmitted by aphids can break down host defenses, potentially weakening these associations and increasing the host breadth of their vectors. Here, we assessed vector host preference and performance in an experiment with five pea aphid colonies with distinct host associations, five host species, and two viruses (Pea enation mosaic virus [PEMV] and Bean leafroll virus [BLRV]). Our prediction was that pea aphids would have higher performance on their inferior hosts and be less discriminating among hosts when those are infected, compared with noninfected hosts. Our results show that aphid performance, measured as the rate of increase on hosts, differed among hosts based on their host association, and relative performance among hosts was indeed altered by virus infection. Aphid preferences, measured using a choice bioassay in which aphids could settle on any of the five host species, also differed among aphid colonies but was not affected by virus infection status. We also did not find that infection status of hosts altered the performance of aphids on their preferred host. Furthermore, virus infection did not disrupt preference performance relationships for the aphids. Overall, our study suggests that host preference and performance are deeply ingrained in pea aphids, and that host associations may be robust to the presence of host-manipulating plant pathogens.

Diversity and Traits of Multiple Biotic Stressors Elicit Differential Defense Responses in Legumes

In agroecosystems, plants frequently confront multiple biotic stressors, including herbivores and pathogens. The nature of these interactions plays a crucial role in mediating the activation of plant defense mechanisms. However, induction of plant chemical defenses has been more well studied than the induction of physical defenses. Here, we assessed the physical and chemical defense responses of pea (Pisum sativum) plants after exposure to three stressors: a vector herbivore (pea aphid, Acrythosiphon pisum), a non-vector herbivore (pea leaf weevil, Sitona lineatus), and a virus (Pea enation mosaic virus, PEMV). We used various histochemical staining techniques show that viruliferous A. pisum (transmitting PEMV) strongly induced callose deposition (aniline blue staining) and antioxidant-mediated defenses (DAB and NBT staining) in peas, primarily through accumulating reactive oxygen species (ROS). High-throughput phenotyping showed that viruliferous aphids reduced plant photosynthetic efficiency, but plants infected with PEMV had increased cell death (trypan blue staining). However, herbivory by aphids and weevils did not strongly induce defenses in peas, even though weevil feeding significantly reduced pea leaf area. These results show that not all herbivores induce strong defensive responses, and plant responses to vector species depends on their virus infection status. More broadly, our results indicate that variable stressors differentially regulate various plant responses through intricate chemical and physical defense pathways.

Crystal structure of a cap-independent translation enhancer RNA.

In eukaryotic messengerRNAs, the 5' cap structure binds to the translation initiation factor 4E to facilitate early stages of translation. Although many plant viruses lack the 5' cap structure, some contain cap-independent translation elements (CITEs) in their 3' untranslated region. The PTE (Panicum mosaic virus translation element) class of CITEs contains a G-rich asymmetric bulge and a C-rich helical junction that were proposed to interact via formation of a pseudoknot. SHAPE analysis of PTE homologs reveals a highly reactive guanosine residue within the G-rich region proposed to mediate eukaryotic initiation factor 4E (eIF4E) recognition. Here we have obtained the crystal structure of the PTE from Pea enation mosaic virus 2 (PEMV2) RNA in complex with our structural chaperone, Fab BL3-6. The structure reveals that the G-rich and C-rich regions interact through a complex network of interactions distinct from those expected for a pseudoknot. The motif, which contains a short parallel duplex, provides a structural mechanism for how the guanosine is extruded from the core stack to enable eIF4E recognition. Homologous PTE elements harbor a G-rich bulge and a three-way junction and exhibit covariation at crucial positions, suggesting that the PEMV2 tertiary architecture is conserved among these homologs.

Molecular characteristics and pathogenicity of infectious cDNA clones of the Chinese isolates of pea enation mosaic virus 1 and pea enation mosaic virus 2

The pea enation mosaic disease (PEMD) causes significant yield losses worldwide. PEMD is caused by two taxonomically unrelated but symbiotic viruses, pea enation mosaic virus 1 (PEMV-1) and pea enation mosaic virus 2 (PEMV-2). In this study, the complete genomes of four isolates of PEMV-1 and PEMV-2 from Yunnan Province of China were determined and analyzed. The four isolates of PEMV-1 or PEMV-2 shared ≥ 98.7% nucleotide sequence identities at whole genome level. The four Chinese PEMV-1 isolates were most closely related to PEMV-1-Ramsey-1 with nucleotide sequence identities of 97.8–98.0%, while the four Chinese PEMV-2 isolates were most closely related to PEMV-2-DSMZ PV-0088, with nucleotide sequence identities of 94.3–94.5%. The two full-length infectious cDNA clones of the Yunnan-Dali isolates of the two viruses (PEMV-1-YDL and PEMV-2-YDL) were constructed. Co-inoculation of PEMV-1-YDL and PEMV-2-YDL in pea seedlings caused systemic infection, with typical enation and mosaic symptoms on new leaves after inoculation, while the inoculated Nicotiana benthamiana plants showed curling symptoms. Inoculation of pea seedlings with either PEMV-1-YDL or PEMV-2-YDL did not cause obvious symptoms on the new leaves. It has been known that PEMV-2 helps the systemic infection of PEMV-1. However, our results showed that PEMV-1-YDL alone could cause a systemic infection. The pathogenicity of the two infectious clones was tested on different pea cultivars, and some cultivars were PEMD-tolerant. This is the first report of the infectious clones constructed from the Chinese isolates of PEMV-1 and PEMV-2. The current approach will facilitate determination of the synergistic relationship between PEMV-1 and PEMV-2 and their interactions with the host plant by reverse genetic analysis.

Conserved Structure Associated with Different 3'CITEs Is Important for Translation of Umbraviruses.

The cap-independent translation of plus-strand RNA plant viruses frequently depends on 3' structures to attract translation initiation factors that bind ribosomal subunits or bind directly to ribosomes. Umbraviruses are excellent models for studying 3' cap-independent translation enhancers (3'CITEs), as umbraviruses can have different 3'CITEs in the central region of their lengthy 3'UTRs, and most also have a particular 3'CITE (the T-shaped structure or 3'TSS) near their 3' ends. We discovered a novel hairpin just upstream of the centrally located (known or putative) 3'CITEs in all 14 umbraviruses. These CITE-associated structures (CASs) have conserved sequences in their apical loops and at the stem base and adjacent positions. In 11 umbraviruses, CASs are preceded by two small hairpins joined by a putative kissing loop interaction (KL). Converting the conserved 6-nt apical loop to a GNRA tetraloop in opium poppy mosaic virus (OPMV) and pea enation mosaic virus 2 (PEMV2) enhanced translation of genomic (g)RNA, but not subgenomic (sg)RNA reporter constructs, and significantly repressed virus accumulation in Nicotiana benthamiana. Other alterations throughout OPMV CAS also repressed virus accumulation and only enhanced sgRNA reporter translation, while mutations in the lower stem repressed gRNA reporter translation. Similar mutations in the PEMV2 CAS also repressed accumulation but did not significantly affect gRNA or sgRNA reporter translation, with the exception of deletion of the entire hairpin, which only reduced translation of the gRNA reporter. OPMV CAS mutations had little effect on the downstream BTE 3'CITE or upstream KL element, while PEMV2 CAS mutations significantly altered KL structures. These results introduce an additional element associated with different 3'CITEs that differentially affect the structure and translation of different umbraviruses.

Risk assessment for non‐crop hosts of pea enation mosaic virus and the aphid vector Acyrthosiphon pisum

Abstract Viral insect‐borne plant pathogens have devastating impacts in agroecosystems. Vector‐borne pathogens are often transmitted by generalist insects that move between non‐crop and crop hosts. Insect vectors can have wide diet breadths, but it is often unknown which hosts serve as pathogen reservoirs and which non‐crop host harbours the highest density of vectors. In the Pacific Northwest USA, the pea aphid (Acyrthosiphon pisum) is a key virus vector in pulse crops. Despite pea aphid having a large number of potential non‐crop plant hosts occuring in the region, no reservoir has yet been identified for the economically‐costly pathogen Pea Enation Mosaic Virus (PEMV). We addressed these issues by linking field surveys of an aphid vector and plant virus with statistical models to develop risk assessments for common non‐crop legumes; in 2018, we completed a 65‐site survey where aphids were surveyed in weedy legumes within and outside dry pea fields. We quantified the abundance of pea aphids on 17 hosts, and plant tissue was tested for PEMV. Relatively high densities of A. pisum were found in habitats dominated by hairy vetch (Vicia villosa), which was the only legume other than cultivated dry pea where PEMV was detected. Our results indicate that V. villosa is a key alternative host for PEMV, and that pest management practices in this region should consider the distribution and abundance of this weedy host in viral disease mitigation efforts for pulses.

Physical Properties of Potyvirus on Chilli (Capsicum annuum) of Doon Valley in Uttarakhand

A virus is transmitted mechanically in the plant host. Potyviridae is a family of Potyvirus,consisting of a positive-sense RNA genome and filamentous particles. It has a single typeof protein coat. Mostly, this virus is identified in Leguminosae and Solanaceae family. Theplant virus is transmitted by mechanical inoculation and aphids. Chilli, cucurbits, potatoes,peas, tomatoes, and other plants were infected with the bean yellow mosaic virus, pea mosaicvirus, bean yellow mosaic virus, sugarcane mosaic virus, and peanut mosaic virus. Thermalinactivation point, dilution endpoint, longevity in vitro, and longevity in vivo were used toidentify the physical properties of a potyvirus. Several isolates mild and severe were collectedin the chilli plant. The infected plant developed necrotic leaf spots and chlorotic symptoms.

THE ROLE OF BIOTIC FACTORS IN <i>LUPINUS POLYPHYLLUS </i>LINDL.(FABACEAE)INVASIVENESS LIMITATION

The article presents experimental data on the species composition of fungal and viral pathogens in the conditions of the secondary range of Lupinus polyphyllus . Tobacco mosaic virus, Bean yellow mosaic virus, Bean common mosaic virus and Pea enation mosaic virus were diagnosed on Lupinus polyphyllus for the first time. The issues related to the peculiarities of the adaptability of viruses to invasive plant species are discussed. The preventive role of vectors ( Aphididae ) in the expansion of pathogens and the widening of the spectrum of host plants (susceptible species) is emphasized. Interaction with vectors, including their non-specific species, is one of the mechanisms of virus adaptability, their expansion into new regions and the formation of new pathosystems with invasive plant species. It is concluded that based on the analysis of trophic connections of vectors, it is possible to prognosticate the search for the most effective variants of harmful organisms for the biocontrol of L. polyphyllus .

Novel 3' Proximal Replication Elements in Umbravirus Genomes.

The 3' untranslated regions (UTRs) of positive-strand RNA plant viruses commonly contain elements that promote viral replication and translation. The ~700 nt 3'UTR of umbravirus pea enation mosaic virus 2 (PEMV2) contains three 3' cap-independent translation enhancers (3'CITEs), including one (PTE) found in members of several genera in the family Tombusviridae and another (the 3'TSS) found in numerous umbraviruses and several carmoviruses. In addition, three 3' terminal replication elements are found in nearly every umbravirus and carmovirus. For this report, we have identified a set of three hairpins and a putative pseudoknot, collectively termed "Trio", that are exclusively found in a subset of umbraviruses and are located just upstream of the 3'TSS. Modification of these elements had no impact on viral translation in wheat germ extracts or in translation of luciferase reporter constructs in vivo. In contrast, Trio hairpins were critical for viral RNA accumulation in Arabidopsis thaliana protoplasts and for replication of a non-autonomously replicating replicon using a trans-replication system in Nicotiana benthamiana leaves. Trio and other 3' terminal elements involved in viral replication are highly conserved in umbraviruses possessing different classes of upstream 3'CITEs, suggesting conservation of replication mechanisms among umbraviruses despite variation in mechanisms for translation enhancement.

Complex and simple translational readthrough signals in pea enation mosaic virus 1 and potato leafroll virus, respectively.

Different essential viral proteins are translated via programmed stop codon readthrough. Pea enation mosaic virus 1 (PEMV1) and potato leafroll virus (PLRV) are related positive-sense RNA plant viruses in the family Solemoviridae, and are type members of the Enamovirus and Polerovirus genera, respectively. Both use translational readthrough to express a C-terminally extended minor capsid protein (CP), termed CP-readthrough domain (CP-RTD), from a viral subgenomic mRNA that is transcribed during infections. Limited incorporation of CP-RTD subunits into virus particles is essential for aphid transmission, however the functional readthrough structures that mediate CP-RTD translation have not yet been defined. Through RNA solution structure probing, RNA secondary structure modeling, site-directed mutagenesis, and functional in vitro and in vivo analyses, we have investigated in detail the readthrough elements and complex structure involved in expression of CP-RTD in PEMV1, and assessed and deduced a comparatively simpler readthrough structure for PLRV. Collectively, this study has (i) generated the first higher-order RNA structural models for readthrough elements in an enamovirus and a polerovirus, (ii) revealed a stark contrast in the complexity of readthrough structures in these two related viruses, (iii) provided compelling experimental evidence for the strict requirement for long-distance RNA-RNA interactions in generating the active readthrough signals, (iv) uncovered what could be considered the most complex readthrough structure reported to date, that for PEMV1, and (v) proposed plausible assembly pathways for the formation of the elaborate PEMV1 and simple PLRV readthrough structures. These findings notably advance our understanding of this essential mode of gene expression in these agriculturally important plant viruses.

Impact of Pea necrotic yellow dwarf virus (PNYDV) on nodulation, N2 fixation and yield in faba bean (Vicia faba, L.)

Pea necrotic yellow dwarf virus (PNYDV) is a novel nanovirus in Europe, affecting various grain legumes. The impact of PNYDV on nodulation, symbiotic N2 fixation and yield parameters in faba bean (Vicia faba L.) was investigated at controlled conditions in the greenhouse (2017), on-farm in natural infection foci (2018, 2019) and in a small-scale field experiment (2020). In the latter, the standard variety ‘Fuego’ was compared with the variety ‘GL Sunrise’ in early and late infections. In addition, the analogous effects for Pea enation mosaic virus (PEMV) were investigated under greenhouse conditions and the naturally occurring virus spectrum was recorded on-farm and in the field experiment. Results showed a much more severe impact of PNYDV than PEMV on nodulation, leghemoglobin status, N2 fixation and finally yield, especially in early infections. Although ‘GL Sunrise’ was rated for a less symptomatic field performance toward PNYDV than ‘Fuego’, it showed a similar susceptibility in our field experiment where PNYDV was artificially inoculated to individual plants. Further research on the effect of plant varieties on susceptibility toward PNYDV infection and its spread in single or co-infection mainly with PEMV as a function of climate change acting upon their common aphid vector is required.

Genomic characterization of a new enamovirus infecting common bean.

A novel enamovirus was identified in common bean plants with disease symptoms. Its genome of 5,781 nucleotides (nt) contains five open reading frames. This virus and other members of the genus Enamovirus share 50.4-68.4% nucleotide sequence identity in the complete genome and 19.9-51.9% amino acid sequence identity in the P0 protein, 24.9-52.5% in P1, 33.4-62.9% in P1-P2, 30.6-81.1% in P3, and 32.3-74.2% in P3-P5. Phylogenetic analysis showed that the virus is most closely related to alfalfa enamovirus 1 and pea enation mosaic virus 1 in the genus Enamovirus of the family Solemoviridae. These results suggest that this virus, tentatively named "bean enamovirus 1", should be classified as a member of a new species in the genus Enamovirus.

Manipulation of Vector Host Preference by Pathogens: Implications for Virus Spread and Disease Management.

Some plant pathogens manipulate the behavior and performance of their vectors, potentially enhancing pathogen spread. The implications are evolutionary and epidemiological but also economic for pathogens that cause disease in crops. Here we explore with models the effects of vector manipulation on crop yield loss to disease and on the economic returns for vector suppression. We use two frameworks, one that simulates the proportional occurrence of the pathogen in the vector population with the option to eliminate vectors by a single insecticidal treatment, and one that includes vector population dynamics and the potential for multiple insecticidal sprays in a season to suppress vectors. We parameterize the models with published data on vector manipulation, crop yields as affected by the age of the plant at infection, commodity prices and costs of vector control for three pathosystems. Using the first framework, maximum returns for treating vectors are greater with vector manipulation than without it by approximately US$10 per acre (US$24.7/ha) in peas infected by Pea enation mosaic virus and Bean leaf roll virus, and approximately US$50 per acre (US$124/ha) for potatoes infected by Potato leaf roll virus. Using the second framework, maximum returns for controlling the psyllid vectors of Candidatus Liberibacter solanacearum are 50% greater (approximately US$400/acre, US$988/ha) but additional returns for multiple weekly sprays diminish more with vector manipulation than without it. These results suggest that the economics of vector manipulation can be substantial and provide a framework that can inform management decisions.

Quail pea mosaic virus

This datasheet on Quail pea mosaic virus covers Identity, Distribution, Hosts/Species Affected.

Integrating High throughput Sequencing into Survey Design Reveals Turnip Yellows Virus and Soybean Dwarf Virus in Pea (Pisum Sativum) in the United Kingdom.

There is only limited knowledge of the presence and incidence of viruses in peas within the United Kingdom, therefore high-throughput sequencing (HTS) in combination with a bulk sampling strategy and targeted testing was used to determine the virome in cultivated pea crops. Bulks of 120 leaves collected from twenty fields from around the UK were initially tested by HTS, and presence and incidence of virus was then determined using specific real-time reverse-transcription PCR assays by testing smaller mixed-bulk size samples. This study presents the first finding of turnip yellows virus (TuYV) in peas in the UK and the first finding of soybean dwarf virus (SbDV) in the UK. While TuYV was not previously known to be present in UK peas, it was found in 13 of the 20 sites tested and was present at incidences up to 100%. Pea enation mosaic virus-1, pea enation mosaic virus-2, pea seed-borne mosaic virus, bean yellow mosaic virus, pea enation mosaic virus satellite RNA and turnip yellows virus associated RNA were also identified by HTS. Additionally, a subset of bulked samples were re-sequenced at greater depth to ascertain whether the relatively low depth of sequencing had missed any infections. In each case the same viruses were identified as had been identified using the lower sequencing depth. Sequencing of an isolate of pea seed-borne mosaic virus from 2007 also revealed the presence of TuYV and SbDV, showing that both viruses have been present in the UK for at least a decade, and represents the earliest whole genome of SbDV from Europe. This study demonstrates the potential of HTS to be used as a surveillance tool, or for crop-specific field survey, using a bulk sampling strategy combined with HTS and targeted diagnostics to indicate both presence and incidence of viruses in a crop.

Development of a highly sensitive and rapid detection method for Pea enation mosaic virus using loop-mediated isothermal amplification assay

The Pea enation mosaic virus (PEMV) has infected plants in the family Leguminosae such as pea, chickpea, faba bean, and lentil plants worldwide that the virus can be transmitted by sap, aphids, and seeds. Among the damages that PEMV disease cause in plants are reduced crop productivity, severely misshapen pods, wart-like outgrowths or proliferation on the surface. Previously, enzyme-linked immunosorbent assay (ELISA), reverse transcription (RT)-nested polymerase chain reaction (PCR), and real-time PCR had been used to detect PEMV. However, these methods are time-consuming and require specific equipments. For this reason, the development of a highly specific and sensitive detection method has become necessary. In this study, a new method for PEMV-1 using the loop-mediated isothermal amplification (LAMP) assay has been developed with specific primer sets as inner- and outer primers. Results showed PEMV-1 has been successfully detected that LAMP could confirm a diluted PEMV-1 up to 10−6 cDNA. LAMP is about 10,000 times more sensitive than the RT-nested PCR and/or real-time PCR. Moreover, the processing time of the LAMP was decreased 3 h than RT-nested PCR. Although future validation will be required to confirm enablement in the field area, this study provides a valuable method to identify PEMV-1 that could offer some advantages including rapid detection, high specificity and high sensitivity than others.