1887

Abstract

Black queen cell virus (BQCV) is a severe threat to the honeybee () worldwide. Although several BQCV strains have been reported in China, the molecular basis for BQCV pathogenicity has not been well understood. Thus, a reverse genetic system of BQCV is required for studying viral replication and its pathogenic mechanism. Here, the complete genome sequence of BQCV was obtained from honeybees using reverse transcription PCR (RT-PCR), namely a BQCV China-GS1 strain (KY741959). Then, a phylogenetic tree was built to analyse the genetic relationships among BQCV strains from different regions. Our results showed that the BQCV China-GS1 contained two ORFs, consistent with the known reference strains, except for the BQCV China-JL1 strain (KP119603). Furthermore, the infectious clone of BQCV was constructed based on BQCV China-GS1 using a low copy vector pACYC177 and gene recombination. Due to the lack of culture cells for bee viruses, we infected the healthy bees with infectious clone of BQCV, and the rescued BQCV resulted in the recovery of recombinant virus, which induced higher mortality than those of the control group. Immune response after inoculated with BQCV further confirmed that the infectious clone of BQCV caused the cellular and humoral immune response of honeybee (). In conclusion, the full nucleotide sequence of BQCV China-GS1 strain was determined, and the infectious clone of BQCV was constructed in this study. These data will improve the understanding of pathogenesis and the host immune responses to viral infection.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001770
2022-08-10
2024-04-18
Loading full text...

Full text loading...

References

  1. Fürst MA, McMahon DP, Osborne JL, Paxton RJ, Brown MJF. Disease associations between honeybees and bumblebees as a threat to wild pollinators. Nature 2014; 506:364–366 [View Article] [PubMed]
    [Google Scholar]
  2. Oldroyd BP. What’s killing American honey bees?. PLoS Biol 2007; 5:e168 [View Article] [PubMed]
    [Google Scholar]
  3. Amiri E, Meixner M, Nielsen SL, Kryger P. Four Categories of Viral Infection Describe the Health Status of Honey Bee Colonies. PLoS One 2015; 10:e0140272 [View Article] [PubMed]
    [Google Scholar]
  4. Zhang X, He SY, Evans JD, Pettis JS, Yin GF et al. New evidence that deformed wing virus and black queen cell virus are multi-host pathogens. J Invertebr Pathol 2012; 109:156–159 [View Article] [PubMed]
    [Google Scholar]
  5. Reddy KE, Noh JH, Choe SE, Kweon CH, Yoo MS et al. Analysis of the complete genome sequence and capsid region of black queen cell viruses from infected honeybees (Apis mellifera) in Korea. Virus Genes 2013; 47:126–132 [View Article] [PubMed]
    [Google Scholar]
  6. Grozinger CM, Flenniken ML. Bee Viruses: Ecology, Pathogenicity, and Impacts. Annu Rev Entomol 2019; 64:205–226 [View Article] [PubMed]
    [Google Scholar]
  7. Beaurepaire A, Piot N, Doublet V, Antunez K, Campbell E et al. Diversity and global distribution of viruses of the western honey bee, Apis mellifera. Insects 2020; 11:E239 [View Article]
    [Google Scholar]
  8. Ai H, Yan X, Han R. Occurrence and prevalence of seven bee viruses in Apis mellifera and Apis cerana apiaries in China. J Invertebr Pathol 2012; 109:160–164 [View Article] [PubMed]
    [Google Scholar]
  9. Yuan C, Jiang X, Liu M, Yang S, Deng S et al. An investigation of honey bee viruses prevalence in managed honey bees (Apis mellifera and Apis cerana) undergone colony decline. TOMICROJ 2021; 15:58–66 [View Article]
    [Google Scholar]
  10. Murray EA, Burand J, Trikoz N, Schnabel J, Grab H et al. Viral transmission in honey bees and native bees, supported by a global black queen cell virus phylogeny. Environ Microbiol 2019; 21:972–983 [View Article] [PubMed]
    [Google Scholar]
  11. Waller GD. Contemporary Queen Rearing. Bulletin of the Entomological Society of America 1980; 26:409–410 [View Article]
    [Google Scholar]
  12. Mondet F, de Miranda JR, Kretzschmar A, Le Conte Y, Mercer AR. On the front line: quantitative virus dynamics in honeybee (Apis mellifera L.) colonies along a new expansion front of the parasite Varroa destructor. PLoS Pathog 2014; 10:e1004323 [View Article] [PubMed]
    [Google Scholar]
  13. Spurny R, Přidal A, Pálková L, Kiem HKT, de Miranda JR et al. Virion Structure of Black Queen Cell Virus, a Common Honeybee Pathogen. J Virol 2017; 91:e02100-16 [View Article] [PubMed]
    [Google Scholar]
  14. Leat N, Ball B, Govan V, Davison S. Analysis of the complete genome sequence of black queen-cell virus, a picorna-like virus of honey bees. J Gen Virol 2000; 81:2111–2119 [View Article] [PubMed]
    [Google Scholar]
  15. Tapaszti Z, Forgách P, Kovágó C, Topolska G, Nowotny N et al. Genetic analysis and phylogenetic comparison of Black queen cell virus genotypes. Vet Microbiol 2009; 139:227–234 [View Article] [PubMed]
    [Google Scholar]
  16. Yang Q, Song Z-Y, Feng X, Zhang J, Zheng Y et al. Analysis of the complete genome sequence of black queen cell virus JL1 from infected honeybees in China. Bull Entomol Res 2016; 106:561–568 [View Article] [PubMed]
    [Google Scholar]
  17. Mookhploy W, Kimura K, Disayathanoowat T, Yoshiyama M, Hondo K et al. Capsid Gene Divergence of Black Queen Cell Virus Isolates in Thailand and Japan Honey Bee Species. J Econ Entomol 2015; 108:1460–1464 [View Article] [PubMed]
    [Google Scholar]
  18. Iwamoto T, Okinaka Y, Mise K, Mori K-I, Arimoto M et al. Identification of host-specificity determinants in betanodaviruses by using reassortants between striped jack nervous necrosis virus and sevenband grouper nervous necrosis virus. J Virol 2004; 78:1256–1262 [View Article] [PubMed]
    [Google Scholar]
  19. Al Naggar Y, Paxton RJ. Mode of Transmission Determines the Virulence of Black Queen Cell Virus in Adult Honey Bees, Posing a Future Threat to Bees and Apiculture. Viruses 2020; 12:E535 [View Article] [PubMed]
    [Google Scholar]
  20. Brutscher LM, Daughenbaugh KF, Flenniken ML. Antiviral Defense Mechanisms in Honey Bees. Curr Opin Insect Sci 2015; 10:71–82 [View Article] [PubMed]
    [Google Scholar]
  21. Yang S, Zhao H, Deng Y, Deng S, Wang X et al. A Reverse Genetics System for the Israeli Acute Paralysis Virus and Chronic Bee Paralysis Virus. Int J Mol Sci 2020; 21:1742 [View Article] [PubMed]
    [Google Scholar]
  22. Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol Biol Evol 2016; 33:1870–1874 [View Article] [PubMed]
    [Google Scholar]
  23. Tamura K, Nei M. Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol Biol Evol 1993; 10:512–526 [View Article] [PubMed]
    [Google Scholar]
  24. Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA et al. Clustal W and Clustal X version 2.0. Bioinformatics 2007; 23:2947–2948 [View Article] [PubMed]
    [Google Scholar]
  25. Huang S, Li J, Zhang Y, Li Z, Evans JD et al. A novel method for the detection and diagnosis of virus infections in honey bees. J Virol Methods 2021; 293:114163 [View Article] [PubMed]
    [Google Scholar]
  26. Doublet V, Paxton RJ, McDonnell CM, Dubois E, Nidelet S et al. Brain transcriptomes of honey bees (Apis mellifera) experimentally infected by two pathogens: Black queen cell virus and Nosema ceranae. Genom Data 2016; 10:79–82 [View Article] [PubMed]
    [Google Scholar]
  27. Biesmeijer JC, Roberts SPM, Reemer M, Ohlemüller R, Edwards M et al. Parallel declines in pollinators and insect-pollinated plants in Britain and the Netherlands. Science 2006; 313:351–354 [View Article] [PubMed]
    [Google Scholar]
  28. Kerr PJ, Rogers MB, Fitch A, Depasse JV, Cattadori IM et al. Genome scale evolution of myxoma virus reveals host-pathogen adaptation and rapid geographic spread. J Virol 2013; 87:12900–12915 [View Article] [PubMed]
    [Google Scholar]
  29. Dalmon A, Gayral P, Decante D, Klopp C, Bigot D et al. Viruses in the Invasive Hornet Vespa velutina. Viruses 2019; 11:10–41 [View Article]
    [Google Scholar]
  30. McMahon DP, Fürst MA, Caspar J, Theodorou P, Brown MJF et al. A sting in the spit: widespread cross-infection of multiple RNA viruses across wild and managed bees. J Anim Ecol 2015; 84:615–624 [View Article] [PubMed]
    [Google Scholar]
  31. Ye S, Xia H, Dong C, Cheng Z, Xia X et al. Identification and characterization of Iflavirus 3C-like protease processing activities. Virology 2012; 428:136–145 [View Article] [PubMed]
    [Google Scholar]
  32. Xia X, Zhou B, Wei T. Complete genome of Chinese sacbrood virus from Apis cerana and analysis of the 3C-like cysteine protease. Virus Genes 2015; 50:277–285 [View Article] [PubMed]
    [Google Scholar]
  33. Painter MM, Morrison JH, Zoecklein LJ, Rinkoski TA, Watzlawik JO et al. Antiviral Protection via RdRP-Mediated Stable Activation of Innate Immunity. PLoS Pathog 2015; 11:e1005311 [View Article] [PubMed]
    [Google Scholar]
  34. Harami GM, Gyimesi M, Kovács M. From keys to bulldozers: expanding roles for winged helix domains in nucleic-acid-binding proteins. Trends Biochem Sci 2013; 38:364–371 [View Article] [PubMed]
    [Google Scholar]
  35. Domeier ME, Morse DP, Knight SW, Portereiko M, Bass BL et al. A link between RNA interference and nonsense-mediated decay in Caenorhabditis elegans. Science 2000; 289:1928–1931 [View Article] [PubMed]
    [Google Scholar]
  36. Linder P, Daugeron MC. Are DEAD-box proteins becoming respectable helicases?. Nat Struct Biol 2000; 7:97–99 [View Article] [PubMed]
    [Google Scholar]
  37. Chen YP, Pettis JS, Corona M, Chen WP, Li CJ et al. Israeli acute paralysis virus: epidemiology, pathogenesis and implications for honey bee health. PLoS Pathog 2014; 10:e1004261 [View Article] [PubMed]
    [Google Scholar]
  38. Zotti M, Dos Santos EA, Cagliari D, Christiaens O, Taning CNT et al. RNA interference technology in crop protection against arthropod pests, pathogens and nematodes. Pest Manag Sci 2018; 74:1239–1250 [View Article] [PubMed]
    [Google Scholar]
  39. Yang D, Xu X, Zhao H, Yang S, Wang X et al. Diverse Factors Affecting Efficiency of RNAi in Honey Bee Viruses. Front Genet 2018; 9:384 [View Article] [PubMed]
    [Google Scholar]
  40. Boncristiani HF, Evans JD, Chen Y, Pettis J, Murphy C et al. In vitro infection of pupae with Israeli acute paralysis virus suggests disturbance of transcriptional homeostasis in honey bees (Apis mellifera). PLoS One 2013; 8:e73429 [View Article] [PubMed]
    [Google Scholar]
  41. Xia X, Mao Q, Wang H, Zhou B, Wei T. Replication of Chinese sacbrood virus in primary cell cultures of Asian honeybee (Apis cerana). Arch Virol 2014; 159:3435–3438 [View Article] [PubMed]
    [Google Scholar]
  42. Benjeddou M, Leat N, Allsopp M, Davison S. Development of infectious transcripts and genome manipulation of Black queen-cell virus of honey bees. J Gen Virol 2002; 83:3139–3146 [View Article] [PubMed]
    [Google Scholar]
  43. Wang G-H, Tsai T-H, Kui C-C, Cheng C-Y, Huang T-L et al. Analysis of bioavailable toluene by using recombinant luminescent bacterial biosensors with different promoters. J Biol Eng 2021; 15:2 [View Article] [PubMed]
    [Google Scholar]
  44. Yang L, Qiu LM, Fang Q, Stanley DW, Ye GY. Cellular and humoral immune interactions between Drosophila and its parasitoids. Insect Sci 2021; 28:1208–1227 [View Article] [PubMed]
    [Google Scholar]
  45. Small C, Paddibhatla I, Rajwani R, Govind S. An introduction to parasitic wasps of Drosophila and the antiparasite immune response. J Vis Exp 2012e3347 [View Article] [PubMed]
    [Google Scholar]
  46. Hillyer JF. Insect immunology and hematopoiesis. Dev Comp Immunol 2016; 58:102–118 [View Article] [PubMed]
    [Google Scholar]
  47. Wang S, Chen G, Lin Z, Wu Y, Hu F et al. Occurrence of multiple honeybee viruses in the ectoparasitic mites Varroa spp. in Apis cerana colonies. J Invertebr Pathol 2019; 166:107225 [View Article] [PubMed]
    [Google Scholar]
  48. Chen G, Wu Y, Deng J, Wen Z, Wang S et al. Seasonal variation of viral infections between the eastern honey bee (Apis cerana) and the western honey bee (Apis mellifera). Microbiologyopen 2021; 10:e1162 [View Article] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001770
Loading
/content/journal/jgv/10.1099/jgv.0.001770
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error