Infection study of Bombyx mori macula-like virus (BmMLV) using a BmMLV-negative cell line and an infectious cDNA clone

Iwanaga, Masako; Hitotsuyama, Tomoyuki; Katsuma, Susumu; Ishihara, Genki; Daimon, Takaaki; Shimada, Toru; Imanishi, Shigeo; Kawasaki, Hideki

doi:10.1016/j.jviromet.2011.11.016

Cited by 22 publications

(47 citation statements)

References 20 publications

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“…The vectors of Maculavirus are unknown [1]. Recently, a macula-like virus BmMLV was reported which is infectious to and replicable in B. mori-derived cells [19], [22]. So far no mosquitoes have been reported as vectors of plant viruses.…”

Section: Discussionmentioning

confidence: 99%

Genomic Characterization of a Novel Virus of the Family Tymoviridae Isolated from Mosquitoes

Wang¹,

Lv²,

Zhai³

et al. 2012

PLoS ONE

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BackgroundThe family Tymoviridae comprises three plant virus genera, including Tymovirus, Marafivirus, and Maculavirus, which are found in most parts of the world and cause severe agricultural losses. We describe a putatively novel member of the family Tymoviridae, which is isolated from mosquitoes (Culex spp.), referred to as CuTLV.Methods and ResultsThe CuTLV was isolated by cell culture, which replicates and causes cytopathic effects in Aedes albopictus C6/36 cells, but not in mammalian BHK-21 or Vero cells. The complete 6471 nucleotide sequence of CuTLV was determined. The genome of CuTLV is predicted to contain three open reading frames (ORFs). The largest ORF1 is 5307 nucleotides (nt) in length and encodes a putative polypeptide of 1769 amino acids (aa), which contains the conserved motifs for the methyltransferase (MTR), Tymovirus endopeptidase (PRO), helicase (HEL), and RNA-dependent RNA polymerase (RdRp) of the replication-associated proteins (RPs) of positive-stranded RNA viruses. In contrast, the ORF1 sequence does not contain the so-called “tymobox” or “marafibox”, the conserved subgenomic RNA promoter present in all tymoviruses or marafiviruses, respectively. ORF2 and ORF3 putatively encode a 248-aa coat protein (CP) and a proline-rich 149-aa polypeptide. The whole genomic nucleotide identity of CuTLV with other members of family Tymoviridae ranged from 46.2% (ChiYMV) to 52.4% (GFkV). Phylogenetic analysis based on the putative RP and CP genes of CuTLV demonstrated that the virus is most closely related to viruses in the genus Maculavirus.ConclusionsThe CuTLV is a novel virus related to members of the family Tymoviridae, with molecular characters that are distinct from those of tymoviruses, marafiviruses, and other maculaviruses or macula-like viruses. This is the first report of the isolation of a Tymoviridae-like virus from mosquitoes. Further investigations are required to clarify the origin, replication strategy, and the public health or agricultural importance of the CuTLV.

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Section: Discussionmentioning

confidence: 99%

Genomic Characterization of a Novel Virus of the Family Tymoviridae Isolated from Mosquitoes

Wang¹,

Lv²,

Zhai³

et al. 2012

PLoS ONE

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“…The use of these cell lines has led to the discovery of persistent infections of RNA viruses [7] [8] [9]. For example, a nodavirus was found in High Five cells [7] that were infected with a recombinant baculovirus containing a hepatitis E capsid gene and enhanced nodavirus proliferation was observed in infected cells.…”

Section: Introductionmentioning

confidence: 99%

Complete study demonstrating the absence of rhabdovirus in a distinct Sf9 cell line

Hashimoto¹,

Macri²,

Srivastava³

et al. 2017

PLoS ONE

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A putative novel rhabdovirus (SfRV) was previously identified in a Spodoptera frugiperda cell line (Sf9 cells [ATCC CRL-1711 lot 58078522]) by next generation sequencing and extensive bioinformatic analysis. We performed an extensive analysis of our Sf9 cell bank (ATCC CRL-1711 lot 5814 [Sf9L5814]) to determine whether this virus was already present in cells obtained from ATCC in 1987. Inverse PCR of DNA isolated from Sf9 L5814 cellular DNA revealed integration of SfRV sequences in the cellular genome. RT-PCR of total RNA showed a deletion of 320 nucleotides in the SfRV RNA that includes the transcriptional motifs for genes X and L. Concentrated cell culture supernatant was analyzed by sucrose density gradient centrifugation and revealed a single band at a density of 1.14 g/ml. This fraction was further analysed by electron microscopy and showed amorphous and particulate debris that did not resemble a rhabdovirus in morphology or size. SDS-PAGE analysis confirmed that the protein composition did not contain the typical five rhabdovirus structural proteins and LC-MS/MS analysis revealed primarily of exosomal marker proteins, the SfRV N protein, and truncated forms of SfRV N, P, and G proteins. The SfRV L gene fragment RNA sequence was recovered from the supernatant after ultracentrifugation of the 1.14 g/ml fraction treated with diethyl ether suggesting that the SfRV L gene fragment sequence is not associated with a diethyl ether resistant nucleocapsid. Interestingly, the 1.14 g/ml fraction was able to transfer baculovirus DNA into Sf9L5814 cells, consistent with the presence of functional exosomes. Our results demonstrate the absence of viral particles in ATCC CRL-1711 lot 5814 Sf9 cells in contrast to a previous study that suggested the presence of infectious rhabdoviral particles in Sf9 cells from a different lot. This study highlights how cell lines with different lineages may present different virosomes and therefore no general conclusions can be drawn across Sf9 cells from different laboratories.

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“…Tymoviridae are positive-strand RNA viruses without an envelope. Tymoviruses infect mainly plants but they have also been found in insects, such as, bees (NCBI txid:1,682,186) [24,25], silkworms (288,456 and 2,065,033) [26], mites (1,682,187 and 1,005,993) [25], and mosquitoes (1,236,047) [27].…”

Section: Lampyris Noctiluca Macula-like Virusmentioning

confidence: 99%

Identification and characterisation of common glow-worm RNA viruses

et al. 2020

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The common glow-worms (Lampyris noctiluca) are best known for emission of green light by their larvae and sexually active adult females. However, both their DNA and RNA viruses remain unknown. Glow-worms are virologically interesting, as they are non-social and do not feed as adults, and hence their viral transmission may be limited. We identified viral sequences from 11 different virus taxa by the RNA-sequencing of two Finnish populations of adult glow-worms. The viruses represent nine different virus families and have negative, positive, or double-stranded RNA genomes. We also found a complete retroviral genome. Similar viral sequences were found from the sequencing data of common eastern firefly of North America, a species belonging to the same family (Lampyridae) as that of the common glow-worm. On average, an individual glow-worm had seven different RNA virus types and most of them appeared to establish a stable infection since they were found from glow-worms during two consecutive years. Here we present the characterization of load, prevalence, and interactions for each virus. Most of the glow-worm RNA viruses seem to be transmitted vertically, which may reflect the biology of glow-worms as non-social capital breeders, i.e., they invest stored resources in reproduction.

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Infection study of Bombyx mori macula-like virus (BmMLV) using a BmMLV-negative cell line and an infectious cDNA clone

Cited by 22 publications

References 20 publications

Genomic Characterization of a Novel Virus of the Family Tymoviridae Isolated from Mosquitoes

Genomic Characterization of a Novel Virus of the Family Tymoviridae Isolated from Mosquitoes

Complete study demonstrating the absence of rhabdovirus in a distinct Sf9 cell line

Identification and characterisation of common glow-worm RNA viruses

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