Abstract
Totiviridae, a viral family of double-stranded RNA (dsRNA) viruses, contain a single dsRNA genome 4.6–7.0 kb in length. Totiviridae were initially only known to infect fungi and other eukaryotes as well as plants, but an increase in totiviruses has been detected in insects, mosquitoes, and bats. Here, we describe the isolation and characterization of a strain belonging to the family Totiviridae isolated from Culex tritaeniorhynchus in Kenli, China, in 2016. We isolated a totivirus from field-collected mosquitoes in China by cell culture in Aedes albopictus C6/36 cells, identified the virus by morphological observation and complete genome sequencing, and characterized it by phylogenetic analysis. Transmission electron microscopy identified icosahedral, non-enveloped virus particles with a mean diameter of 35–40 nm. The genome was 7612 bp in length, including two open reading frames (ORFs). ORF1 (5058 nt) encodes the capsid protein, while ORF2 (2216 nt) encodes the viral RNA-dependent RNA polymerase (RdRp). Nucleotide and amino acid homology analysis of isolate showed higher levels of sequence identity with isolate CTV_NJ2 (China, 2010) with 94.87% nucleic acid identity and 97.32% amino acid identity. The isolate was designated C. tritaeniorhynchus totivirus KL (CTV-KL). This is the first identification of a totivirus in a C. tritaeniorhynchus in northern China. Analysis of the virus’s morphology, characteristic and genome organization will further enrich our understanding of the molecular and biological characteristics of dsRNA Totiviridae viruses.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- ORFs:
-
Open reading frames
- RdRp:
-
RNA-dependent RNA polymerase
- CTV:
-
Culex tritaeniorhynchus totivirus
- CPEs:
-
Cytopathic effects
- dsDNA:
-
Double-stranded DNA
- ssDNA:
-
Single-stranded DNA
- dsRNA:
-
Double-stranded RNA
- ssRNA+:
-
Positive single-stranded RNA
- ssRNA−:
-
Negative single-stranded RNA
- ssRNA-RT:
-
Single-stranded retro-transcribing RNA
- dsDNA-RT:
-
Double-stranded retro-transcribing DNA
References
Baltimore D (1971) Expression of animal virus genomes. Bacteriol Rev 35:235–241
de Lima JGS, Teixeira DG, Freitas TT, Lima J, Lanza DCF (2019) Evolutionary origin of 2A-like sequences in Totiviridae genomes. Virus Res 259:1–9. https://doi.org/10.1016/j.virusres.2018.10.011
Ghabrial SA (2008) Totiviruses. In: Mahy BWJ, van Regenmortel MHV (eds) Encyclopedia of virology, vol 5, 3rd edn. Academic Press/Elsevier, San Diego, pp 163–174
Ghabrial SA, Nibert ML (2009) Victorivirus, a new genus of fungal viruses in the family Totiviridae. Adv Virol 154:373–379. https://doi.org/10.1007/s00705-008-0272-x
Goodman RP, Ghabrial SA, Fichorova RN, Nibert ML (2011) Trichomonasvirus: a new genus of protozoan viruses in the family Totiviridae. Adv Virol 156:171–179. https://doi.org/10.1007/s00705-010-0832-8
Hardy JL, Houk EJ, Kramer LD, Reeves WC (1983) Intrinsic factors affecting vector competence of mosquitoes for arboviruses. Annu Rev Entomol 28:229–262. https://doi.org/10.1146/annurev.en.28.010183.001305
Huang Y et al (2018) Discovery of two novel totiviruses from Culex tritaeniorhynchus classifiable in a distinct clade with arthropod-infecting viruses within the family Totiviridae. Adv Virol 163:2899–2902. https://doi.org/10.1007/s00705-018-3871-1
Isawa H et al (2011) Identification and molecular characterization of a new nonsegmented double-stranded RNA virus isolated from Culex mosquitoes in Japan. Virus Res 155:147–155. https://doi.org/10.1016/j.virusres.2010.09.013
Katoh K, Toh H (2008) Recent developments in the MAFFT multiple sequence alignment program. Br Bioinform 9:286–298. https://doi.org/10.1093/bib/bbn013
King AMQ et al (2018) Changes to taxonomy and the International Code of Virus Classification and Nomenclature ratified by the International Committee on Taxonomy of Viruses (2018). Adv Virol 163:2601–2631. https://doi.org/10.1007/s00705-018-3847-1
Kobayashi S, Lauwereyns J, Koizumi M, Sakagami M, Hikosaka O (2002) Influence of reward expectation on visuospatial processing in macaque lateral prefrontal cortex. J Neurophysiol 87:1488–1498. https://doi.org/10.1152/jn.00472.2001
Kondo H et al (2016) Reprint of “Sequence and phylogenetic analyses of novel totivirus-like double-stranded RNAs from field-collected powdery mildew fungi”. Virus Res 219:39–50. https://doi.org/10.1016/j.virusres.2016.05.011
Kuwata R, Isawa H, Hoshino K, Sasaki T, Kobayashi M, Maeda K, Sawabe K (2015) Analysis of mosquito-borne flavivirus superinfection in Culex tritaeniorhynchus (Diptera: Culicidae) cells persistently infected with culex flavivirus (Flaviviridae). J Med Entomol 52:222–229. https://doi.org/10.1093/jme/tju059
Li WJ et al (2010) Mosquitoes and mosquito-borne arboviruses in the Qinghai–Tibet plateau-focused on the Qinghai area, China. Am J Trop Med Hyg 82:705–711. https://doi.org/10.4269/ajtmh.2010.09-0649
Lu BL (1997a) Fauna Sinica, Insecta vol 8, Diptera: Culicidae 1. Science Press, Beijing
Lu BL (1997b) Fauna Sinica, Insecta vol 9, Diptera: Culicidae II. Science Press, Beijing
Lu Z et al (2009) Tahyna virus and human infection, China. Emerg Infect Dis 15:306–309
Poulos BT, Tang KF, Pantoja CR, Bonami JR, Lightner DV (2006) Purification and characterization of infectious myonecrosis virus of penaeid shrimp. J Gen Virol 87:987–996. https://doi.org/10.1099/vir.0.81127-0
Read DA et al (2019) Diversity and distribution of maize-associated totivirus strains from Tanzania. Virus Genes. https://doi.org/10.1007/s11262-019-01650-6
Shapiro A, Green T, Rao S, White S, Carner G, Mertens PP, Becnel JJ (2005) Morphological and molecular characterization of a Cypovirus (Reoviridae) from the mosquito Uranotaenia sapphirina (Diptera: Culicidae). J Virol 79:9430–9438. https://doi.org/10.1128/JVI.79.15.9430-9438.2005
Strauss JH, Strauss EG (1994) The alphaviruses: gene expression, replication, and evolution. Microbiol Rev 58:491–562
Sun X et al (2009) Distribution of arboviruses and mosquitoes in northwestern Yunnan Province, China. Vector Borne Zoonotic Dis 9:623–630. https://doi.org/10.1089/vbz.2008.0145
Tang KF, Pantoja CR, Poulos BT, Redman RM, Lightnere DV (2005) In situ hybridization demonstrates that Litopenaeus vannamei, L. stylirostris and Penaeus monodon are susceptible to experimental infection with infectious myonecrosis virus (IMNV). Dis Aquat Organ 63:261–265. https://doi.org/10.3354/dao063261
van Cleef KW et al (2014) Mosquito and Drosophila entomobirnaviruses suppress dsRNA- and siRNA-induced RNAi. Nucleic Acids Res 42:8732–8744. https://doi.org/10.1093/nar/gku528
Wu Q, Luo Y, Lu R, Lau N, Lai EC, Li WX, Ding SW (2010) Virus discovery by deep sequencing and assembly of virus-derived small silencing RNAs. Proc Natl Acad Sci USA 107:1606–1611. https://doi.org/10.1073/pnas.0911353107
Yang J et al (2011) Unbiased parallel detection of viral pathogens in clinical samples by use of a metagenomic approach. J Clin Microbiol 49:3463–3469. https://doi.org/10.1128/JCM.00273-11
Yang X, Zhang Y, Ge X, Yuan J, Shi Z (2012) A novel totivirus-like virus isolated from bat guano. Adv Virol 157:1093–1099. https://doi.org/10.1007/s00705-012-1278-y
Zhai Y et al (2010) Isolation and full-length sequence analysis of Armigeres subalbatustotivirus, the first totivirus isolate from mosquitoes representing a proposed novel genus (Artivirus) of the family Totiviridae. J Gen Virol 91:2836–2845. https://doi.org/10.1099/vir.0.024794-0
Acknowledgements
We thank the staff of the Kenli County Centers for Disease Control and Prevention for assistance with the mosquito collection.
Funding
This work was supported by National Science and Technology Major Project (2017ZX10104001); National Natural Science Foundation of China (31900156); National key research and development project (2016YFC1200905); the funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Author information
Authors and Affiliations
Contributions
FL and HW were major contributors in writing the manuscript. WZ and AX collected the mosquitoes sample. SF, FL, WZ, QW, YH, and WL performed virus isolation. FL sequenced the genome of CTV-KL. JS performed the virus electron microscopy. FL, JD, and ZW did the Bioinformatics analysis. HW, LZ, and GL conceived the study and drafted the manuscript. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no competing interests.
Ethics approval and consent to participate
Not applicable.
Consent for publication
Not applicable.
Availability of supporting data
Not applicable.
Additional information
Communicated by Erko Stackebrandt.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Li, F., Du, J., Wu, Z. et al. Identification and genetic analysis of a totivirus isolated from the Culex tritaeniorhynchus in northern China. Arch Microbiol 202, 807–813 (2020). https://doi.org/10.1007/s00203-019-01788-9
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00203-019-01788-9