Alfalfa enation virus

Lenardon Search for more papers by this author. Affiliations Authors and Affiliations N. Bejerman C. Nome F. Add to favorites Download Citations Track Citations. Metrics Downloaded 16 times. Alfalfa mosaic virus AMV : genetic diversity and a new natural host. Geographical distribution, genetic studies and vector transmission of alfalfa enamovirus-1 infecting alfalfa crop in Argentina.

Diversity and epidemiology of plant rhabdoviruses. Morphological changes, alteration of photosynthetic parameters and chlorophyll production induced by infection with alfalfa dwarf virus in Medicago sativa plants.

Characterisation of a novel nucleorhabdovirus infecting alfalfa Medicago sativa. Bejerman, N. Redefining the Medicago sativa alphapartitiviruses genome sequences. Virus Res. Complete genome sequence and integrated protein localization and interaction map for alfalfa dwarf virus, which combines properties of both cytoplasmic and nuclear plant rhabdoviruses. Virology , — Complete genome sequence of a new enamovirus from argentina infecting alfalfa plants showing dwarfism symptoms.

First report of a rhabdovirus infecting alfalfa in argentina. Plant Dis. Genome characterization of an argentinean isolate of alfalfa leaf curl virus. Bekal, S. A novel flavivirus in the soybean cyst nematode. Bernardo, P. Geometagenomics illuminates the impact of agriculture on the distribution and prevalence of plant viruses at the ecosystem scale.

ISME J. Molecular characterization and prevalence of two capulaviruses: alfalfa leaf curl virus from France and Euphorbia caput-medusae latent virus from South Africa. Blouin, A. A new virus discovered by immunocapture of double-stranded RNA, a rapid method for virus enrichment in metagenomic studies. Boccardo, G. Cryptic plant viruses. Brough, R. The historical diffusion of alfalfa.

Candresse, T. Appearances can be deceptive: revealing a hidden viral infection with deep sequencing in a plant quarantine context.

PLoS One 9:e Davoodi, Z. The westward journey of alfalfa leaf curl virus. Viruses Debat, H. Donaire, L. Deep-sequencing of plant viral small RNAs reveals effective and widespread targeting of viral genomes. Dong, W.

PLoS One 3:e Elena, S. The games plant viruses play. Filloux, D. Metagenomics approaches based on virion-associated nucleic acids vana : an innovative tool for assessing without a priori viral diversity of plants. Methods Mol.

Viral metagenomics approaches for high-resolution screening of multiplexed arthropod and plant viral communities. Frate, C. Fujimoto, Y. Complete genome sequence of lychnis mottle virus isolated in Japan.

Genome Announc. Gaafar, Y. Characterization of a novel nucleorhabdovirus infecting alfalfa Medicago sativa. Gilbert, K. Hiding in plain sight: new virus genomes discovered via a systematic analysis of fungal public transcriptomes. PLoS One e Guo, Z.

First report of alfalfa leaf curl virus infecting alfalfa Medicago sativa in China. Hampton, R. Evidence suggesting identity between alfalfa latent andpea streak viruses. Phytopathology Han, S. Nucleotide sequence and taxonomy of cycas necrotic stunt virus. Hanada, K. Cycas necrotic stunt virus isolated from gladiolus plants in Japan. Ho, T. Development of a virus detection and discovery pipeline using next generation sequencing. Virology —, 54— Huang, X. Genome Res. Jiang, P. Identification of emerging viral genomes in transcriptomic datasets of alfalfa Medicago sativa L.

Identification of the coding-complete genome of cycas necrotic stunt virus in transcriptomic data sets of alfalfa Medicago sativa. Jones, S. Viral diagnostics in plants using next generation sequencing: computational analysis in practice. Joshi, N. Julier, B. Legumes in Cropping Systems. Wallingford: CABI. Kim, H. Gene , 7— Koenig, R. Brunt, K. Crabtree, M. Dallwitz, A. Gibbs, L. Watson, and E.

Kreuze, J. Complete viral genome sequence and discovery of novel viruses by deep sequencing of small RNAs: a generic method for diagnosis, discovery and sequencing of viruses. Virology , 1—7. Krupovic, M. Plant viruses of the Amalgaviridae family evolved via recombination between viruses with double-stranded and negative-strand RNA genomes. Kusunoki, M.

Cycas necrotic stunt virus, a new member of nepoviruses found in Cycas revoluta host range, purification, serology and some other properties. Lambert, C. Considerations for optimization of high-throughput sequencing bioinformatics pipelines for virus detection.

Viruses E Lamichhane, J. Synergisms between microbial pathogens in plant disease complexes: a growing trend. Langmead, B. Fast gapped-read alignment with Bowtie 2.

Methods 9, — Lauber, C. Discovery of highly divergent lineages of plant-associated astro-like viruses sheds light on the emergence of potyviruses. Leclant, F. Li, Y. VIP: an integrated pipeline for metagenomics of virus identification and discovery. Lim, S. Genomic detection and molecular characterization of two distinct isolates of cycas necrotic stunt virus from Paeonia suffruticosa and Daphne odora.

Virus Genes 55, — Liou, M. Complete nucleotide sequence and genome organization of a Cactus virus X strain from Hylocereus undatus Cactaceae. Ma, Y. Phytovirome analysis of wild plant populations: comparison of double-stranded RNA and virion-associated nucleic acid metagenomic approaches.

Maina, S. Metagenomic analysis reveals a nearly complete genome sequence of alfalfa mosaic virus from a field pea in Australia. Malvick, D. Maree, H. Application of HTS for routine plant virus diagnostics: state of the art and challenges. Martelli, G. Family flexiviridae: a case study in virion and genome plasticity. Martin, R. Blueberry latent virus: an amalgam of the Partitiviridae and Totiviridae.

There are no known resistant commercial cultivars. Controls rely on isolation of pea plantings from virus sources.

Bean yellow mosaic virus BYMV , also called bean virus 2 in older literature, includes pea mosaic virus strains, which some authors treat separately. BYMV is widely distributed in the state and is transmitted by at least 20 aphid species in a nonpersistent manner. This virus is also discussed under the virus diseases affecting bean. The symptoms of BYMV depend upon the strain of virus involved.

The pea strain, for example, causes a very bright yellow mosaic fig. Resistance for BYMV has been identified and extends to the Perfection types used for both canning and freezing. Resistance is conditioned by a recessive factor, which, when it occurs in a heterozygous condition, is strongly influenced by temperature.

Seed transmission is absent or very rare in pea and several other legumes. Aphid control may help to reduce the rate of spread in susceptible cultivars. Clover yellow vein virus CYVV causes a serious disease of pea. A severe strain can cause intense veinal chlorosis or severe yellowing, followed by apical necrosis and premature death. Plants are usually stunted, and pods show some deformation.

The virus is transmitted in nature by at least four aphid species including the pea aphid in a nonpersistent manner. This virus also naturally infects bean and cucurbits, as well as many cultivated and wild legumes, and ornamentals such as wild violet.

Symptoms of CYVV infection in pea consist of veinal chlorosis, mosaic, curling, and apical rosetting, with suppression of pod size and number fig. Cultivars resistant to BYMV are also resistant to this virus. Isolate plantings from potential virus reservoirs or take steps to remove these sources before the crop is planted. When ultrathin sections of embedded symptomatic N. Figure 2 a shows two virus particles in epidermal cells. The transversely cut particle seems to be complete with attachment of glycoproteins visible lower arrow, left hand side.

Figure 2 a upper arrow, right hand side seems to show two longitudinally particles appearing blunt end to blunt end and thus looking like a larger particle. Both epidermal and mesophyl cells were infected. Electron micrographs of thin sections of AaNV-infected N. The cell wall W , chloroplast with starch granules Cl , nucleus Nu , mitochondrion Mt , vacuole Va , virus budding Vb and viroplasm Vp are indicated.

In infected cells, the shape of the nucleus can change to a more condensed circular or even a distorted shape Fig. In heavily infected cells, not only the nuclear compartments were affected but also chloroplast were deformed Fig.

In the nuclei, granular areas distinct from heterochromatin were found representing putative virus replication sites known as viroplasms Vp. Adjacent to them virus particles could be found Fig. In Fig. In addition, vesicles or virus buddings Vb containing one or more complete viruses were visualized around the nucleus and in the cytoplasm of infected cells Fig. Figure 2 d also shows virus particles budding from the inner nuclear envelope in the perinuclear space.

A total of 1,, reads were generated from the MiSeq sequencing. After quality trimming and size filtering, 1,, quality-filtered reads were used for normalisation and de novo assembly. From the 23, assembled contigs, a contig of 13, nucleotides showed The sequences of the two ends were assembled to the contig and the full-length genome sequence was determined as 13, bases in length with 29, mapped reads, The sequencing dataset generated in this study is available from the corresponding author upon request.

A pairwise nucleotide sequence alignment of the novel genome to selected rhabdoviruses and a phylogenetic tree was generated. ClustalW pairwise analysis showed that the AaNV sequence has This clustering was supported by a neighbour joining tree of the L protein amino acid sequences of selected members of the family Rhabdoviridae Fig.

Unrooted neighbour-joining phylogenetic tree [Genetic distance model Jones-Taylor-Thornton JTT model and bootstrap replications] based on the amino acid sequence alignment of the L protein of selected members of different genera of the family Rhabdoviridae.

AaNV indicated by a blue solid diamond shape. Six putative open reading frames ORFs were identified in the antigenomic sense based on the genome organisation described for other nucleorhabdoviruses; nucleocapsid N , phosphoprotein P , putative cell to cell movement protein P3 , matrix protein M , glycoprotein G and RNA-dependent RNA polymerase L. Highly conserved regulatory regions separating the genes were identified.

The presence of a seventh ORF with unknown function U , between M and G was identified as it is flanked by the intergenic region consensus motif. The read map distribution is shown in light blue over the genome. The AaNV protein sizes range from amino acid aa for the U protein to aa for the L protein with molecular masses of The aa sequences identities were between Additionally, the nuclear localisation signals NLS, or a karyophilic domain and the nuclear export signals NES of the proteins were predicted Table 1.

The M protein had the lowest score with 4. The cNLS scores predicted an exclusive nuclear localisation for N, P and L proteins, a partial nuclear localisation for P3 and G proteins, and a nuclear and cytoplasmic localisation for M protein Table 1. Moreover, four of these proteins have a detectable NES Table 1.

Only nucleocapsid structures reacted with antibodies but not complete virions, see Additional file 1 : Figure S3a displaying enriched but undecorated nucleocapsids from the crude sap samples after the preincubation with antiserum immunosorbent step , and Additional file 1 : Figure S3b showing nucleocapsids covered with antibodies after the decoration step.

In a limited host range study, the virus was successfully transmitted to N. Mechanically inoculated N. However, inoculated P. It was not possible to transmit AaNV mechanically to M.

Additionally, mechanical inoculation using fresh material from AaNV-infected V. Photo of AaNV-infected N. Red arrows indicating inoculated leave. The bacilliform appearance of the viral particles observed in infected N. The observed ultra-cellular deformations of nuclei and chloroplast in epidermis and mesophyl cells are in accordance with the distorted phenotype of systemically infected N. The AaNV genome shares As for the third demarcation criterium, AaNV was originally identified in Medicago sativa , an important legume crop.

As a consequence, AaNV should be considered as a new virus species in the Nucleorhabdovirus genus. The predicted features of AaNV proteins are similar to those of related nucleorhabdoviruses. Its predicted protein has an acidic IEP. All the seven transcription units and the leader are predicted to be polyadenylated, but its functionality still needs to be proven. Nucleorhabdoviruses are known to establish virus replication factories in the nuclei of infected plant cells [ 21 ].

All AaNV proteins except U, display predicted mono- or bipartite nuclear localisation sequences NLS suggesting their independent importation into the nucleus. The presence of both the NLSs and the leucine-rich nuclear export signals NESs in N, M and L proteins indicates the ability of these proteins to shuttle between the nucleus and the cytoplasm through coordination of these transport signals.

Although the unknown protein U seems to lack a NLS, the observation that it has an NES suggests its ability to be exported out of the nucleus. AaNV was mechanically transmitted to N. Although it did not show any noticeable or only slight mottling symptoms on P.

Interestingly, the virus could not be mechanically transmitted to M. It is not known if this is due to the serial passaging on N. As a insect vector has not been identified yet, it is unclear how the transmission from M. It is also unknown if this virus still occurs naturally in alfalfa in the area it was originally found, or elsewhere in Europe.

In the present study, we describe a novel nucleorhabdovirus originating from infected M. Since the sequence identity to BCaRV-1, its closest known relative, was only The site of virus maturation was observed by EM in the nucleus of infected cells thus confirming the phylogenetic assignment.

It was possible to transfer AaNV experimentally using mechanical inoculation to N. However, it was not possible to transfer AaNV back to alfalfa by mechanical inoculation. Thus, further research is needed to identify natural vectors of this virus as well as other alternative host plants.

The serological and molecular biological assays developed may aid larger surveys addressing these questions. The dehydration of lucerne Medicago sativa and its potentialities as a concentrated source of ascorbic acid and of carotene for human consumption.