Abstract

BackgroundCylindrospermopsis raciborskii is an invasive filamentous freshwater cyanobacterium, some strains of which produce toxins. Sporadic toxicity may be the result of gene deletion events, the horizontal transfer of toxin biosynthesis gene clusters, or other genomic variables, yet the evolutionary drivers for cyanotoxin production remain a mystery. Through examining the genomes of toxic and non-toxic strains of C. raciborskii, we hoped to gain a better understanding of the degree of similarity between these strains of common geographical origin, and what the primary differences between these strains might be. Additionally, we hoped to ascertain why some cyanobacteria possess the cylindrospermopsin biosynthesis (cyr) gene cluster and produce toxin, while others do not. It has been hypothesised that toxicity or lack thereof might confer a selective advantage to cyanobacteria under certain environmental conditions.ResultsIn order to examine the fundamental differences between toxic and non-toxic C. raciborskii strains, we sequenced the genomes of two closely related isolates, CS-506 (CYN+) and CS-509 (CYN-) sourced from different lakes in tropical Queensland, Australia. These genomes were then compared to a third (reference) genome from C. raciborskii CS-505 (CYN+). Genome sizes were similar across all three strains and their G + C contents were almost identical. At least 2,767 genes were shared among all three strains, including the taxonomically important rpoc1, ssuRNA, lsuRNA, cpcA, cpcB, nifB and nifH, which exhibited 99.8-100% nucleotide identity. Strains CS-506 and CS-509 contained at least 176 and 101 strain-specific (or non-homologous) genes, respectively, most of which were associated with DNA repair and modification, nutrient uptake and transport, or adaptive measures such as osmoregulation. However, the only significant genetic difference observed between the two strains was the presence or absence of the cylindrospermopsin biosynthesis gene cluster. Interestingly, we also identified a cryptic secondary metabolite gene cluster in strain CS-509 (CYN-) and a second cryptic cluster common to CS-509 and the reference strain, CS-505 (CYN+).ConclusionsOur results confirm that the most important factor contributing to toxicity in C. raciborskii is the presence or absence of the cyr gene cluster. We did not identify any other distally encoded genes or gene clusters that correlate with CYN production. The fact that the additional genomic differences between toxic and non-toxic strains were primarily associated with stress and adaptation genes suggests that CYN production may be linked to these physiological processes.

Highlights

  • Cylindrospermopsis raciborskii is an invasive filamentous freshwater cyanobacterium, some strains of which produce toxins

  • The data sets supporting the results presented in this manuscript are available in the following repository: Integrated Microbial Genomes [IMG] repository, unique persistent identifier 12992 and 12991 and NCBI short read archive under accession numbers: SRR1042336 and SRR1041118 for C. raciborskii strains CS-506 and CS509, respectively

  • Genomic structure The genomes of C. raciborskii strains CS-506 and CS509 were sequenced via the bridge amplification method on an Illumina genome analyzer

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Summary

Introduction

Cylindrospermopsis raciborskii is an invasive filamentous freshwater cyanobacterium, some strains of which produce toxins. Some of the toxins produced include cyclic hepatotoxic peptides such as microcystin and nodularin [3], alkaloids such as the cytotoxic cylindrospermopsin (CYN) [4] and the neurotoxic saxitoxin (STX) [5], and organophosphates, such as anatoxin-a(s) [6]. These cyanotoxins, produced by over 40 species from 20 genera of cyanobacteria [7], have adverse health effects on humans and animals, and are a public health and environmental concern [3]. PCC 6506 [12], Anabaena lapponica [13], Lyngbya wollei [14], Umezakia natans [15], Raphidiopsis mediterranea [16], Anabaena bergii [17] and Cylindrospermopsis raciborskii [18]

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