Over 400 cyanobacterial genera have been described up to the present. Since the Cambridge Rules (https://www.iapt-taxon.org/historic/1935.htm: Rendle 1935), a type species (generitype) must be specified at the time of description for a new genus to be validly described. Even though we have entered a time in which sequencing has become practical and widespread, the basic molecular characterization (e.g., 16S rRNA gene sequence) of most cyanobacterial generitypes is still lacking. About 15 cyanobacterial genera were originally described from Scandinavia. Following a field excursion in which the type or syntype localities for the type species of these genera were visited and sampled, we succeeded in finding three type species from their type or syntype localities: Capsosira brebissonii, Stigonema mamillosum, and Paracapsa siderophila. Epitypes for all three generitypes are herein established. Cells or filaments of C. brebissonii and S. mamillosum were isolated and used for single-cell/filament PCR amplification of the 16S rRNA gene and subsequent cloning and sequencing of the PCR amplicons. This allows a firm establishment of reference sequences of these two genera, to which morphologically similar taxa can now be compared. Stigonema and Capsosira are shown herein to be sister to Aetokthonos hydrillicola, a cyanobacterium known to cause avian vacuolar myelinopathy in birds, including bald eagles.

• Klíčová slova
• Capsosira brebissonii, Paracapsa siderophila, Stigonema mamillosum, 16S rRNA gene, 16S–23S ITS, avian vacuolar myelinopathy, cyanobacteria, generitypes, type locality,
• MeSH
• DNA bakterií genetika   MeSH
• fylogeneze MeSH
• RNA ribozomální 16S genetika   analýza   MeSH
• sekvenční analýza DNA MeSH
• sinice * genetika   klasifikace   MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Skandinávie a severské státy MeSH
• Názvy látek
• DNA bakterií MeSH
• RNA ribozomální 16S MeSH

Cyanobacteria are infamous producers of toxins. While the toxic potential of planktonic cyanobacterial blooms is well documented, the ecosystem level effects of toxigenic benthic and epiphytic cyanobacteria are an understudied threat. The freshwater epiphytic cyanobacterium Aetokthonos hydrillicola has recently been shown to produce the "eagle killer" neurotoxin aetokthonotoxin (AETX) causing the fatal neurological disease vacuolar myelinopathy. The disease affects a wide array of wildlife in the southeastern United States, most notably waterfowl and birds of prey, including the bald eagle. In an assay for cytotoxicity, we found the crude extract of the cyanobacterium to be much more potent than pure AETX, prompting further investigation. Here, we describe the isolation and structure elucidation of the aetokthonostatins (AESTs), linear peptides belonging to the dolastatin compound family, featuring a unique modification of the C-terminal phenylalanine-derived moiety. Using immunofluorescence microscopy and molecular modeling, we confirmed that AEST potently impacts microtubule dynamics and can bind to tubulin in a similar matter as dolastatin 10. We also show that AEST inhibits reproduction of the nematode Caenorhabditis elegans. Bioinformatic analysis revealed the AEST biosynthetic gene cluster encoding a nonribosomal peptide synthetase/polyketide synthase accompanied by a unique tailoring machinery. The biosynthetic activity of a specific N-terminal methyltransferase was confirmed by in vitro biochemical studies, establishing a mechanistic link between the gene cluster and its product.

• Klíčová slova
• aetokthonostatin, biosynthesis, cyanotoxin, cytotoxicity, dolastatin,
• MeSH
• Accipitridae * MeSH
• Caenorhabditis elegans MeSH
• ekosystém MeSH
• sinice * genetika   MeSH
• sladká voda MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aetokthonotoxin MeSH Prohlížeč

Cyanobacteria are well known producers of bioactive metabolites, including harmful substances. The recently discovered "eagle killer" neurotoxin aetokthonotoxin (AETX) is produced by the epiphytic cyanobacterium Aetokthonos hydrillicola growing on invasive water thyme (Hydrilla verticillata). The biosynthetic gene cluster of AETX was previously identified from an Aetokthonos strain isolated from the J. Strom Thurmond Reservoir, Georgia, USA. Here, a PCR protocol for easy detection of AETX-producers in environmental samples of plant-cyanobacterium consortia was designed and tested. Three different loci of the AETX gene cluster were amplified to confirm the genetic potential for AETX production, along with two variable types of rRNA ITS regions to confirm the homogeneity of the producer´s taxonomic identity. In samples of Hydrilla from three Aetokthonos-positive reservoirs and one Aetokthonos-negative lake, the PCR of all four loci provided results congruent with the Aetokthonos presence/absence detected by light and fluorescence microscopy. The production of AETX in the Aetokthonos-positive samples was confirmed using LC-MS. Intriguingly, in J. Strom Thurmond Reservoir, recently Hydrilla free, an Aetokthonos-like cyanobacterium was found growing on American water-willow (Justicia americana). Those specimens were positive for all three aet markers but contained only minute amounts of AETX. The obtained genetic information (ITS rRNA sequence) and morphology of the novel Aetokthonos distinguished it from all the Hydrilla-hosted A. hydrillicola, likely at the species level. Our results suggest that the toxigenic Aetokthonos spp. can colonize a broader array of aquatic plants, however the level of accumulation of the toxin may be driven by host-specific interactions such as the locally hyper-accumulated bromide in Hydrilla.

• Klíčová slova
• Aetokthonos, Cyanotoxin, Hydrilla, Justicia, Vacuolar myelinopathy, rRNA ITS,
• MeSH
• chromatografie kapalinová MeSH
• hmotnostní spektrometrie MeSH
• jezera * MeSH
• polymerázová řetězová reakce MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aetokthonotoxin MeSH Prohlížeč