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Статья: First evaluation of the Vairimorpha (Nosema) ceranae genome’s DNA replication genes as targets for RNA interference-mediated suppression of the honey bee Apis mellifera nosemosis (2025)

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In recent decades, infection of the European honey bee Apis mellifera with the highly virulent microsporidium Vairimorpha (Nosema) ceranae has become globally prevalent. It causes serious losses in beekeeping worldwide and requires new approaches to control bee nosemosis. Since this intracellular parasite has retained components of the RNA interference pathway, double-stranded RNA (dsRNA) treatment of insects may be effective in controlling V. ceranae infection. Inhibition of microsporidia growth in bees fed with 1.8 or even 0.04 μg dsRNA per ml of sugar syrup has been reported in the literature. Considering the crucial role of the genome’s DNA replication machinery for any cell, we synthetized in vitro dsRNA fragments of four V. ceranae genes encoding two subunits (delta and epsilon) of DNA polymerase, helicase and topoisomerase II and fed them to the infected bees with a relatively low dose of 1 µg per ml of sugar syrup. Surprisingly, PCR and qPCR analyses of V. ceranae growth in the midgut of dsRNA-treated insects at 7- and 12-days post-infection revealed neither inhibition of microsporidia growth nor downregulation of target genes. It is worth noting that we collected worker bees of different ages directly from a hive, to simulate the conditions during colony treatment in apiaries. At the same time, newly emerged insects reared in the laboratory were used in the successful experiments mentioned above. This suggests that bee housing conditions as well as gut content may affect the efficiency of RNA interference and requires further increase in dsRNA doses or their mixing with nanoparticle carriers.

Ключевые фразы: honey bee, apis mellifera, microsporidia, vairimorpha ceranae, nosemosis control, rna interference, double-stranded rna, dna replication genes, gene silencing

Автор (ы): Долгих Владимир Валентинович, IGNATIEVA A.N., RUMIANTSEVA A.S., TIMOFEEV S.A., FADEEV R.R., BAYAZYT K.D.K.

Журнал: PROTISTOLOGY

Предпросмотр статьи

Идентификаторы и классификаторы

SCI: Биология
УДК: 57. Биологические науки

Для цитирования:

ДОЛГИХ В. В., IGNATIEVA A.N., RUMIANTSEVA A.S., TIMOFEEV S.A., FADEEV R.R., BAYAZYT K.D.K. FIRST EVALUATION OF THE VAIRIMORPHA (NOSEMA) CERANAE GENOME’S DNA REPLICATION GENES AS TARGETS FOR RNA INTERFERENCE-MEDIATED SUPPRESSION OF THE HONEY BEE APIS MELLIFERA NOSEMOSIS // PROTISTOLOGY. 2025. № 1, ТОМ 19

Текстовый фрагмент статьи

Список литературы

1. Ahmad S., Jamil M., Jaworski C.C. and Luo Y.P. 2023. Double-stranded RNA degrading nuclease affects RNAi efficiency in the melon fly. J. Pest. Sci. 97: 397-409. DOI: 10.1007/s10340-023-01637-1
2. Arjunan N., Thiruvengadam V. and Sushil S.N. 2024. Nanoparticle-mediated dsRNA delivery for precision insect pest control: a comprehensive review. Mol. Biol. Rep. 51: 355. DOI: 10.1007/s11033-023-09187-6
3. Bernhardt H.S. and Tate W.P. 2012. Primordial soup or vinaigrette: did the RNA world evolve at acidic pH? Biol. Direct. 7: 4. DOI: 10.1186/1745-6150-7-4
4. Bjornson S. and Oi D. 2014. Microsporidia biological control agents and pathogens of beneficial insects. In: Microsporidia: pathogens of opportunity (Eds: Weiss L.M. and Becnel J.J.). Wiley-Blackwell, pp. 635-670.
5. Cantwell G.E. 1970. Standard methods for counting Nosema Spores. Am. Bee J. 110: 222-223.
6. Desai S.D., Eu Y.J., Whyard S. and Currie R.W. 2012. Reduction in deformed wing virus infection in larval and adult honey bees (Apis mellifera L.) by double-stranded RNA ingestion. Insect. Mol. Biol. 21: 446-455. DOI: 10.1111/j.1365-2583.2012.01150.x
7. Fadeev R.R., Kudryavtseva Yu.S., Bayazyt K-D.K., Shuhalova A.G. and Dolgikh V.V. 2024. The optimized method to isolate dsRNA synthesized in Escherichia coli HT115(DE3). Agricultural Biology. 59: 460-472. DOI: 10.15389/agrobiology.2024.3.460eng
8. Fries I., Feng F., da Silva A., Slemenda S.B. and Pieniazek N.J. 1996. Nosema ceranae n. sp. (Microspora, Nosematidae), morphological and molecular characterization of a microsporidian parasite of the Asian honey bee Apis cerana (Hymenoptera, Apidae). Eur. J. Protistol. 32: 356-365.
9. Gisder S. and Genersch E. 2015. Identification of candidate agents active against N. ceranae infection in honey bees: establishment of a medium throughput screening assay based on N. ceranae infected cultured cells. PLoS One. 10: e0117200. DOI: 10.1371/journal.pone.0117200
10. Guan R., Chu D., Han X., Miao X. and Li H. 2021. Advances in the development of microbial double-stranded RNA production systems for application of RNA interference in agricultural Pest Control. Review Front. Bioeng. Biotechnol. 9: 753790. DOI: 10.3389/fbioe.2021.753790
11. Guan R.B, Li H.C., Fan Y.J., Hu S.R. et al. 2018. A nuclease specific to lepidopteran insects suppresses RNAi. J. Biol. Chem. 293: 6011-6021. DOI: 10.1074/jbc.RA117.001553
12. He N., Zhang Y., Duan X.L., Li J.H. et al. 2021. RNA interference-mediated knockdown of genes encoding spore wall proteins confers protection against Nosema ceranae infection in the European honey bee, Apis mellifera. Microorganisms. 9: 505. DOI: 10.3390/microorganisms9030505 EDN: CYZPFJ
13. Higes M., Martín-Hernández R., Botías C., Bailón E.G., González-Porto A.V. et al. 2009. Honeybee colony collapse due to Nosema ceranae in professional apiaries. Environ. Microbiol. Rep. 1: 110-113. DOI: 10.1111/j.1758-2229.2009.00014.x
14. Higes M., Martín-Hernández R., Garrido- Bailón E. et al. 2008. How natural infection by Nosema ceranae causes honeybee colony collapse. Environ. Microbiol. 10: 2659-2669. DOI: 10.1111/j.1462-2920.2008.01687.x
15. James R.R. and Li Z. 2012. From silkworms to bees: diseases of beneficial Insects. In: Insect Pathology (Eds: Vega F.E. and Kaya H.K.). Elsevier Inc., pp. 429-459.
16. Ignatieva A.N., Timofeev S.A., Tokarev Y.S. and Dolgikh V.V. 2022. Laboratory cultivation of Vairimorpha (Nosema) ceranae (Microsporidia: Nosematidae) in artificially infected worker bees. Insects. 13: 1092. DOI: 10.3390/insects13121092
17. Lang H.Y., Wang H., Wang H.Q., Zhong Z.P. et al. 2023. Engineered symbiotic bacteria interfering redox system inhibit microsporidia parasitism in honeybees. Nat.Commun. 14: 2778. DOI: 10.1038/s41467-023-38498-2
18. Lin-Ling W., Ke-Ping C., Ze Z., Qin Y. et al. 2006. Phylogenetic analysis of Nosema antheraeae (Microsporidia) isolated from Chinese oak silkworm, Antheraea pernyi. J. Eukaryot. Microbiol. 53: 310-313. DOI: 10.1111/j.1550-7408.2006.00106.x
19. Maori E., Paldi N., Shafir S., Kalev H. et al. 2009. IAPV, a bee-affecting virus associated with colony collapse disorder can be silenced by dsRNA ingestion. Insect. Mol. Biol. 18: 55-60. DOI: 10.1111/j.1365-2583.2009.00847.x
20. McGruddy R.A., Smeele Z.E., Manley B., Masucci J.D. et al. 2024. RNA interference as a next-generation control method for suppressing Varroa destructor reproduction in honey bee (Apis mellifera) hives. Pest. Manag. Sci. 80: 4770-4778. DOI: 10.1002/ps.8193
21. Martín-Hernández R., Bartolomé C., Chejanovsky N., Le Conte Y. et al. 2018. Nosema ceranae in Apis mellifera: a 12 years postdetection perspective. Environ. Microbiol. 20: 1302-1329. DOI: 10.1111/1462-2920.14103
22. Ndikumana S., Pelin A., Williot A., Sanders J. L. et al. 2017. Genome analysis of Pseudoloma neurophilia: a microsporidian parasite of zebrafish (Danio rerio). J. Eukaryot. Microbiol. 64: 18-30. DOI: 10.1111/jeu.12331
23. Paldi N., Glick E., Oliva M., Zilberberg Y. et al. 2010. Effective gene silencing in a microsporidian parasite associated with honeybee (Apis mellifera) colony declines. Appl. Environ. Microbiol. 76: 5960- 5964. DOI: 10.1128/AEM.01067-10
24. Pereira T.C. and Lopes-Cendes I. 2013. Medical applications of RNA interference (RNAi). BMC Proc. 7: K21. DOI: 10.1186/1753-6561-7-S2-K21
25. Qi Y., Wang C., Lang H., Wang Y. et al. 2024. Liposome-based RNAi delivery in honeybee for inhibiting parasite Nosema ceranae. Synth. Syst. Biotechnol. 9: 853-860. DOI: 10.1016/j.synbio.2024.07.003
26. Rodríguez-García C., Evans J.D., Li W., Branchiccela B. et al. 2018. Nosemosis control in European honey bees, Apis mellifera, by silencing the gene encoding Nosema ceranae polar tube protein 3. J. Exp. Biol. 221: jeb184606. DOI: 10.1242/jeb.184606
27. Rodrigues T.B., Mishra S.K., Sridharan K., Barnes E.R. et al. 2021. First sprayable double-stranded RNA-based biopesticide product targets proteasome subunit beta type-5 in Colorado potato beetle (Leptinotarsa decemlineata). Front. Plant. Sci. 12: 728652. DOI: 10.3389/fpls.2021.728652
28. Romeis J. and Widmer F. 2020. Assessing the risks of topically applied dsRNA-based products to non-target arthropods. Front. Plant Sci. 11: 679. DOI: 10.3389/fpls.2020.00679
29. Schüler V., Liu Y.C., Gisder S., Horchler L. et al. 2023. Significant, but not biologically relevant: Nosema ceranae infections and winter losses of honey bee colonies.Commun. Biol. 6: 229. DOI: 10.1038/s42003-023-04587-7
30. Timofeev S., Tsarev A. Senderskiy I., Rogozhin E. et al. 2019. Efficient transformation of the entomopathogenic fungus Lecanicillium muscarium by electroporation of germinated conidia. Mycoscience. 60: 197-200. DOI: 10.1016/j.myc.2019.02.010
31. Tokarev Y.S., Timofeev S.A., Malysh J.M., Tsarev A.A. et al. 2018. Hexokinase as a versatile molecular genetic marker for microsporidia. Parasitology. 15: 1-7. DOI: 10.1017/S0031182018001737
32. Wang Y., Yan Q., Lan C., Tang T. et al. 2023. Nanoparticle carriers enhance RNA stability and uptake efficiency and prolong the protection against Rhizoctonia solani. Phytopathology Research. 5: 2. DOI: 10.1186/s42483-023-00157-1
33. Zheng H., Powell J.E., Steele M.I., Dietrich C. and Moran N.A. 2017. Honeybee gut microbiota promotes host weight gain via bacterial metabolism and hormonal signaling. Proc. Natl. Acad. Sci. USA. 114: 4775-4780.

Выпуск

№ 1, Том 19 (2025)

Кол-во страниц: 65 страниц

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Characterization of the unusual PII protein with elongated T-loop from Micromonas pusilla (2025)

Авторы: VLASOVA V., LAPINA T., ZALUTSKAYA ZH., CHENG QI., ERMILOVA E.

The PII superfamily consists of widespread signal transduction proteins found in all domains of life. The most conserved PII-interactor across oxygenic phototrophs from cyanobacteria to Archaeplastida is the key enzyme of the ornithine/arginine synthesis pathway, N-acetyl-L-glutamate kinase (NAGK). T-loops represent the major PII-receptor binding element and are involved in the interaction with NAGK. Within the class Mamiellophyceae, only the genus Micromonas contains species with the PII protein. Bioinformatic analysis revealed that the PII protein of Micromonas pusilla (MpPII) has an unusually prolonged T-loop. Here, we performed the coupled enzyme assay and showed that MpPII has no remarkable influence on NAGK activity. An engineered variant of MpPII with deletion of four additional amino acids (AATD) in the T-loop restored the ability of this protein to relieve NAGK from feedback inhibition by arginine in a glutamine-dependent manner. The findings are discussed in the context of unusual plasticity of the PII protein family during the evolution of Archaeplastida.

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