A group of seven bacterial strains producing blue-purple pigmented colonies on R2A agar was isolated from freshwater samples collected in a deglaciated part of James Ross Island and Eagle Island, Antarctica, from 2017-2019. The isolates were psychrophilic, oligotrophic, resistant to chloramphenicol, and exhibited strong hydrolytic activities. To clarify the taxonomic position of these isolates, a polyphasic taxonomic approach was applied based on sequencing of the 16S rRNA, gyrB and lepA genes, whole-genome sequencing, rep-PCR, MALDI-TOF MS, chemotaxonomy analyses and biotyping. Phylogenetic analysis of the 16S rRNA gene sequences revealed that the entire group are representatives of the genus Massilia. The closest relatives of the reference strain P8398T were Massilia atriviolacea, Massilia violaceinigra, Massilia rubra, Massilia mucilaginosa, Massilia aquatica, Massilia frigida, Massilia glaciei and Massilia eurypsychrophila with a pairwise similarity of 98.6-100% in the 16S rRNA. The subsequent gyrB and lepA sequencing results showed the novelty of the analysed group, and the average nucleotide identity and digital DNA-DNA hybridisation values clearly proved that P8398T represents a distinct Massilia species. After all these results, we nominate a new species with the proposed name Massilia antarctica sp. nov. The type strain is P8398T (= CCM 8941T = LMG 32108T).

Central European Institute of Technology Masaryk University Kamenice 5 625 00 Brno Czech Republic

Centrum Algatech Institute of Microbiology Czech Academy of Sciences Opatovický mlýn 379 01 Třeboň Czech Republic

Department of Biomedical Engineering Faculty of Electrical Engineering and Communication Brno University of Technology Technická 12 616 00 Brno Czech Republic

Department of Experimental Biology Czech Collection of Microorganisms Faculty of Science Masaryk University Kamenice 5 625 00 Brno Czech Republic

Department of Physical Geography and Geoecology Faculty of Science Charles University Albertov 6 128 00 Praha Czech Republic

Institut für Mikrobiologie Veterinärmedizinische Universität Wien Veterinärplatz 1 A 1210 Wien Austria

Institute of Bioinformatics Department of Informatics Ludwig Maximilians Universität München Amalienstraße 17 803 33 Munich Germany

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