CYANOPHAGE-INDUCED TRANSCRIPTIONAL CHANGES IN RAPHIDIOPSIS RACIBORSKII

Vaiva Slatkevičiūtė1, 2, 3, Barbara Klimczak4, Gediminas Azlbutas2, Adam Antosiak4, Petras Venckus5, Hanna Zkukava1, Natalija Kazlauskienė1, Sarit Avrani6, Dariusz Dziga4, Sigitas Šulčius1, 2

1 Laboratory of Algology and Microbial Ecology, Nature Research Centre, Lithuania

2 Bioinformatics Department, Nature Research Centre, Lithuania

3 Institute of Biotechnology, Life Science Center, Vilnius University, Lithuania

4 Laboratory of Metabolomics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Poland

5 Institute of Biosciences, Life Sciences Center, Vilnius University, Lithuania

6 Aquatic Microbial Interactions Lab, Department of Evolutionary and Environmental Biology, Faculty of Natural Sciences, University of Haifa, Israel

[email protected]

Raphidiopsis raciborskii is a nitrogen-fixing freshwater cyanobacterium that recently gained attention due to its high invasiveness potential, toxicity, and harmful blooms, which can negatively impact biodiversity, ecosystem functions, and water quality. Despite its ecological significance, factors controlling this species’ occurrence and bloom dynamics remain poorly understood. Cyanophages, viruses that infect cyanobacteria, may play a crucial role in microbial population and community dynamics through host lysis, horizontal gene transfer, and metabolic reprogramming [1,2]. Understanding these interactions in freshwater ecosystems is essential for advancing our knowledge of cyanophage ecology and may provide insights into the mechanisms by which viruses regulate cyanobacterial activity, growth, and transcription. Investigations into transcriptional regulation of host cyanobacteria by viral infection are also crucial for the assessment and development of phage-based technologies for the mitigation of harmful algal blooms.

To investigate the transcriptional response of R. raciborskii to cyanophage infection as well as to establish a transcriptional scheme for cyanophage, we used R. raciborskii strain KLL07 and its lytic cyanophage Cr-LKS4 as a model virus-host system. In this study, control and infection treatments were established by adding either filter-sterilized growth medium or cyanophage suspensions at the multiplicity of infection of three (MOI = 3) in four biological replicates to unialgal yet non-axenic cultures. The experiment lasted 120 hours, with samples collected every 4-8 hours for cell and phage abundance measurements, DNA and RNA sequencing, and proteomic analysis.

We will present results on cyanobacterial and cyanophage dynamics during the infection experiment along with differential gene expression analysis during different infection stages (adsorption, DNA replication and lysis) of R. raciborskii. Additionally, changes in R. raciborskii microbiome composition induced by the cyanophage-mediated lysis of the photosynthetic host will be presented. These findings will contribute to a better understanding of the ecological significance of freshwater cyanophages and their impact on cyanobacteria growth and succession.


[1] D. Kolan et al., “Tradeoffs between phage resistance and nitrogen fixation drive the evolution of genes essential for cyanobacterial heterocyst functionality,” The ISME Journal, vol. 18, no. 1, Jan. 2024, doi: 10.1093/ismejo/wrad008.

[2] D. Morimoto, S. Šulčius, and T. Yoshida, “Viruses of freshwater bloom‐forming cyanobacteria: genomic features, infection strategies and coexistence with the host,” Environmental Microbiology Reports, vol. 12, no. 5, pp. 486–502, Aug. 2020, doi: 10.1111/1758-2229.12872.