Aquatic microalgae produce a variety of toxic secondary metabolites, which are a concern for public health and seafood industries, while also presenting a source of pharmacologically valuable compounds. The present study deals with the physiology and molecular genetics of saxitoxin (STX), a cyanobacterial neurotoxic alkaloid. Ecological and chemical parameters have been investigated for their effects on growth and STX production in the cyanobacterium Cylindrospermopsis raciborskii T3, in order to better understand the physiological responses of this cyanobacterium to the anthropogenic eutrophication of water bodies. The results indicated that phosphate, in particular, had an incremental effect on STX production, as well as promoting the up-regulation of transcription of the STX biosynthetic gene cluster (sxt). High temperature was found to negatively affect growth and STX production in this organism. The effects of the plant hormone, jasmonic acid, were also tested, since it has previously been shown to affect plant alkaloid production. The hypothesised similarity between cyanobacterial and plant secondary metabolism in response to this plant hormone was confirmed in the neurotoxic cyanobacterium, C. raciborskii T3, as well as the non-toxic Anabaena sp. PCC7120. Furthermore, investigation of the sxt gene cluster transcriptional map in C. raciborskii T3 was carried out, with identification of three main polycistronic and one monocistronic transcripts. Promoter regions putatively involved in the regulation of STX production in C. raciborskii T3 were also identified. Transcription factor consensus motifs, the pho boxes, were identified in the main promoter region. These conserved motifs are the binding regions for the transcriptional regulator, PhoB, to the pho regulon genes, involved in phosphate uptake during conditions of its depletion in the environment. Moreover, a genomic region adjacent to the sxt gene cluster in C. raciborskii T3 was identified and characterised, putatively encoding a regulatory two-component system. This system appears to be involved in the sensing of environmental signals, in particular depleted phosphate, while activating the transcription of genes involved in its uptake and transport. The results of this study lead to a greater understanding of the complex factors associated with the regulation of STX biosynthesis and bloom-formation, by the cyanobacterium C. raciborskii T3.

Aquatic microalgae produce a variety of toxic secondary metabolites, which are a concern for public health and seafood industries, while also presenting a source of pharmacologically valuable compounds. The present study deals with the physiology and molecular genetics of saxitoxin (STX), a cyanobacterial neurotoxic alkaloid. Ecological and chemical parameters have been investigated for their effects on growth and STX production in the cyanobacterium Cylindrospermopsis raciborskii T3, in order to better understand the physiological responses of this cyanobacterium to the anthropogenic eutrophication of water bodies. The results indicated that phosphate, in particular, had an incremental effect on STX production, as well as promoting the up-regulation of transcription of the STX biosynthetic gene cluster (sxt). High temperature was found to negatively affect growth and STX production in this organism. The effects of the plant hormone, jasmonic acid, were also tested, since it has previously been shown to affect plant alkaloid production. The hypothesised similarity between cyanobacterial and plant secondary metabolism in response to this plant hormone was confirmed in the neurotoxic cyanobacterium, C. raciborskii T3, as well as the non-toxic Anabaena sp. PCC7120. Furthermore, investigation of the sxt gene cluster transcriptional map in C. raciborskii T3 was carried out, with identification of three main polycistronic and one monocistronic transcripts. Promoter regions putatively involved in the regulation of STX production in C. raciborskii T3 were also identified. Transcription factor consensus motifs, the pho boxes, were identified in the main promoter region. These conserved motifs are the binding regions for the transcriptional regulator, PhoB, to the pho regulon genes, involved in phosphate uptake during conditions of its depletion in the environment. Moreover, a genomic region adjacent to the sxt gene cluster in C. raciborskii T3 was identified and characterised, putatively encoding a regulatory two-component system. This system appears to be involved in the sensing of environmental signals, in particular depleted phosphate, while activating the transcription of genes involved in its uptake and transport. The results of this study lead to a greater understanding of the complex factors associated with the regulation of STX biosynthesis and bloom-formation, by the cyanobacterium C. raciborskii T3.