Journal
TOXINS
Volume 4, Issue 10, Pages 900-912Publisher
MDPI
DOI: 10.3390/toxins4100900
Keywords
16S rRNA gene sequence; anaF gene; anatoxin-a; benthic mats; cyanobacteria; homoanatoxin-a; Phormidium
Categories
Funding
- New Zealand Ministry for Business, Innovation and Employment Cumulative Effects programme [CO1X0803]
- Environment Bay of Plenty
- Hawke's Bay Regional Council
- Horizons Regional Council
- Greater Wellington Regional Council
- Environment Canterbury
- Environment Southland
- Institute of Environmental Science and Research Limited
- Biomolecular Interaction Centre
- Evans Fund
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Benthic Phormidium mats can contain high concentrations of the neurotoxins anatoxin-a and homoanatoxin-a. However, little is known about the co-occurrence of anatoxin-producing and non-anatoxin-producing strains within mats. There is also no data on variation in anatoxin content among toxic genotypes isolated from the same mat. In this study, 30 Phormidium strains were isolated from 1 cm(2) sections of Phormidium-dominated mats collected from three different sites. Strains were grown to stationary phase and their anatoxin-a, homoanatoxin-a, dihydroanatoxin-a and dihydrohomoanatoxin-a concentrations determined using liquid chromatography-mass spectrometry. Each strain was characterized using morphological and molecular (16S rRNA gene sequences) techniques. Eighteen strains produced anatoxin-a, dihydroanatoxin-a or homoanatoxin-a. Strains isolated from each mat either all produced toxins, or were a mixture of anatoxin and non-anatoxin-producing genotypes. Based on morphology these genotypes could not be separated. The 16S rRNA gene sequence comparisons showed a difference of at least 17 nucleotides among anatoxin and non-anatoxin-producing strains and these formed two separate sub-clades during phylogenetic analysis. The total anatoxin concentration among toxic strains varied from 2.21 to 211.88 mg kg(-1) ( freeze dried weight), representing a 100 fold variation in toxin content. These data indicate that both the relative abundance of anatoxin and non-anatoxin-producing genotypes, and variations in anatoxin producing capability, can influence the overall toxin concentration of benthic Phormidium mat samples.
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