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Occurrence of aetokthonotoxin producer in natural samples-A PCR protocol for easy detection
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SYSNO ASEP 0583932 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Occurrence of aetokthonotoxin producer in natural samples-A PCR protocol for easy detection Author(s) Štenclová, Lenka (BC-A) ORCID, RID
Wilde, S. B. (US)
Schwark, M. (DE)
Cullen, J. L. (US)
McWhorter, S. A. (US)
Niedermeyer, T. H. J. (DE)
Henderson, W. M. (US)
Mareš, Jan (BC-A) RID, ORCIDNumber of authors 8 Article number 102425 Source Title Harmful Algae. - : Elsevier - ISSN 1568-9883
Roč. 125, Mar (2023)Number of pages 9 s. Language eng - English Country NL - Netherlands Keywords avian vacuolar myelinopathy ; amyotrophic-lateral-sclerosis ; harmful algal blooms ; cyanobacteria ; Aetokthonos ; Cyanotoxin ; Vacuolar myelinopathy Subject RIV DA - Hydrology ; Limnology OECD category Marine biology, freshwater biology, limnology R&D Projects GC19-21649J GA ČR - Czech Science Foundation (CSF) Method of publishing Limited access Institutional support BC-A - RVO:60077344 UT WOS 000967131700001 EID SCOPUS 85151036490 DOI 10.1016/j.hal.2023.102425 Annotation 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 ampli-fied 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. Workplace Biology Centre (since 2006) Contact Dana Hypšová, eje@eje.cz, Tel.: 387 775 214 Year of Publishing 2024 Electronic address https://doi.org/10.1016/j.hal.2023.102425
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