First report of natural infection of beetroot with <i>Beet soil‐borne virus</i>

Gaafar, Yahya Z. A.; Sieg-Müller, Angelika; Lüddecke, P.; Hartrick, Jonas; Seide, Yvonne; Müller, Jürgen; Maaß, Christina; Schuhmann, S.; Richert-Pöggeler, Katja; Blouin, Arnaud; Massart, Sébastien; Ziebell, Heiko

doi:10.5197/j.2044-0588.2019.040.005

Cited by 7 publications

(5 citation statements)

References 4 publications

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“…Electron microscopy is important for viruses' detection, as it requires no prior knowledge for virus detection [7,8]. Nevertheless, it tends not to be able to identify viruses up to species level as its detection is based on the particle morphology, and often fails to detect low titre viruses or viruses that are phloem-limited and disease-causing entities without protein shells such as viroids or satellite RNAs [9,8,10,11].…”

Section: Introductionmentioning

confidence: 99%

Comparative study on three viral enrichment approaches based on RNA extraction for plant virus/viroid detection using high-throughput sequencing

Gaafar

Ziebell

2020

PLoS ONE

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High-throughput sequencing (HTS) has become increasingly popular as virus diagnostic tool. It has been used to detect and identify plant viruses and viroids in different types of matrices and tissues. A viral sequence enrichment method prior to HTS is required to increase the viral reads in the generated data to ease the bioinformatic analysis of generated sequences. In this study, we compared the sensitivity of three viral enrichment approaches, i.e. double stranded RNA (dsRNA), ribosomal RNA depleted total RNA (ribodepleted totRNA) and small RNA (sRNA) for plant virus/viroid detection, followed by sequencing on MiSeq and NextSeq Illumina platforms. The three viral enrichment approaches used here enabled the detection of all viruses/viroid used in this study. When the data was normalised, the recovered viral/viroid nucleotides and depths were depending on the viral genome and the enrichment method used. Both dsRNA and ribo-depleted totRNA approaches detected a divergent strain of Wuhan aphid virus 2 that was not expected in this sample. Additionally, Vicia cryptic virus was detected in the data of dsRNA and sRNA approaches only. The results suggest that dsRNA enrichment has the highest potential to detect and identify plant viruses and viroids. The dsRNA approach used here detected all viruses/viroid, consumed less time, was lower in cost, and required less starting material. Therefore, this approach appears to be suitable for diagnostics laboratories.

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Section: Introductionmentioning

confidence: 99%

Comparative study on three viral enrichment approaches based on RNA extraction for plant virus/viroid detection using high-throughput sequencing

Gaafar

Ziebell

2020

PLoS ONE

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“…This virus has been regarded as a mild pathogen, and very few studies have focused on its potential economic significance [25]. Several field studies have nevertheless reported association of BSBV with plants showing symptoms [26, 27] and mechanical inoculations of beet resulted in significant reductions of taproot weight [28], advocating altogether for a possible deleterious effect on its host.…”

Section: Introductionmentioning

confidence: 99%

RNA silencing machinery contributes to inability of BSBV to establish infection in Nicotiana benthamiana: evidence from characterization of agroinfectious clones of Beet soil-borne virus

Mahillon

Decroës

Peduzzi

et al. 2021

Journal of General Virology

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Beet soil-borne virus (BSBV) is a sugar beet pomovirus frequently associated with Beet necrotic yellow veins virus, the causal agent of the rhizomania disease. BSBV has been detected in most of the major beet-growing regions worldwide, yet its impact on this crop remains unclear. With the aim to understand the life cycle of this virus and clarify its putative pathogenicity, agroinfectious clones have been engineered for each segment of its tripartite genome. The biological properties of these clones were then studied on different plant species. Local infection was obtained on agroinfiltrated leaves of Beta macrocarpa. On leaves of Nicotiana benthamiana, similar results were obtained, but only when heterologous viral suppressors of RNA silencing were co-expressed or in a transgenic line down regulated for both dicer-like protein 2 and 4. On sugar beet, local infection following agroinoculation was obtained on cotyledons, but not on other tested plant parts. Nevertheless, leaf symptoms were observed on this host via sap inoculation. Likewise, roots were efficiently mechanically infected, highlighting low frequency of root necrosis and constriction, and enabling the demonstration of transmission by the vector Polymyxa betae. Altogether, the entire viral cycle was reproduced, validating the constructed agroclones as efficient inoculation tools, paving the way for further studies on BSBV and its related pathosystem.

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“…While sample EPV_18_002 was asymptomatic apart from slight yellowing, samples EPV_18_003 and EPV_18_004 showed stunting, yellowing, necrosis and severe crinkling (Figs. [1][2][3]. It was possible to mechanically transmit the suspected virus from all three samples to Nicotiana benthamiana; the same systemic symptoms of leaf crinkling, rolling and yellowing appeared seven days post infection on all plants, and the plants died in the second week.…”

Section: Chapter 2: Plant Disease Aetiologymentioning

confidence: 90%

“…Affected plants displayed leaf-rolling, yellowing, and symptoms of severe stunting (Figs. [1][2]. The samples reacted positively when analysed with ELISA using antibodies against PNYDV .…”

Section: Chapter 2: Plant Disease Aetiologymentioning

confidence: 98%

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Plant virus identification and virus-vector-host interactions

Gaafar¹

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BackgroundNucleorhabdoviruses possess bacilliform particles which contain a single-stranded negative-sense RNA genome. They replicate and mature in the nucleus of infected cells. Together with viruses of three other genera of the family Rhabdoviridae, they are known to infect plants and can be transmitted by arthropod vectors, during vegetative propagation, or by mechanical means. In 2010, an alfalfa (Medicago sativa) plant showing virus-like symptoms was collected from Stadl-Paura, Austria and sent to Julius Kühn Institute for analysis. MethodsElectron microscopy (EM) of leaf extracts from infected plants revealed the presence of rhabdovirus-like particles and was further used for ultrastructural analyses of infected plant tissue. Partially-purified preparations of rhabdovirus nucleocapsids were used for raising an antiserum. To determine the virus genome sequence, high throughput sequencing (HTS) was performed. RT-PCR primers were designed to confirm virus infection and to be used as a diagnostic tool. ResultsEM revealed bacilliform virions resembling those of plant-infecting rhabdoviruses. HTS of ribosomal RNA-depleted total RNA extracts revealed a consensus sequence consisting of 13,875 nucleotides (nt) and containing seven open reading frames (ORFs). Homology and phylogenetic analyses suggest that this virus isolate represents a new species of the genus Nucleorhabdovirus (family Rhabdoviridae). Since the virus originated from an alfalfa plant in Austria, the name alfalfa-associated nucleorhabdovirus (AaNV) is proposed. Viroplasms (Vp) and budding virions were observed in the nuclei of infected cells by EM, thus confirming its taxonomic assignment based on sequence data. ConclusionsIn this study, we identified and characterised a new nucleorhabdovirus from alfalfa. It shared only 39.8% nucleotide sequence identity with its closest known relative, black currant-associated rhabdovirus 1. The virus contains an additional open reading frame (accessory gene) with unknown function, located between the matrix protein and the glycoprotein genes. Serological and molecular diagnostic assays were designed for future screening of field samples. Further studies are needed to identify other natural hosts and potential vectors.

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First report of natural infection of beetroot with Beet soil‐borne virus

Cited by 7 publications

References 4 publications

Comparative study on three viral enrichment approaches based on RNA extraction for plant virus/viroid detection using high-throughput sequencing

Comparative study on three viral enrichment approaches based on RNA extraction for plant virus/viroid detection using high-throughput sequencing

RNA silencing machinery contributes to inability of BSBV to establish infection in Nicotiana benthamiana: evidence from characterization of agroinfectious clones of Beet soil-borne virus

Plant virus identification and virus-vector-host interactions

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