Objective: To develop and apply molecular tools to gain comprehensive insight into mechanisms regulating growth, toxicity, and death in the Florida red tide dinoflagellate, Karenia brevis
Background & Results: Our approach is to first to establish a database of expressed genes by high throughput sequencing of cDNA libraries to Karenia brevis under different growth conditions. The expressed sequence database provides the source of sequence information needed to develop DNA microarrays for gene expression profiling, translated peptide libraries for proteomic analysis, and antibodies for characterization of specific gene products. DNA microarrays will be used to identify changes in gene expression under conditions that alter growth or toxicity, characterize the genetic networks controlling these responses, and identify the function of novel genes based on their responses to known stressors. Proteomic analysis is used to identify processes regulated at the post-transcriptional level. Genes involved in key regulatory processes will be fully characterized using in silico and biochemical approaches. A sub-component of this project will attempt to develop transformation and gene silencing (RNAi) protocols to facilitate rigorous analysis of K. brevis gene function. In the final phase of this project, key molecular targets will also assessed for their potential as biomarkers suitable for deployment in remote detection. This project addresses the need for improved understanding of the biological mechanisms controlling HABs as identified in “HARRNESS, 2005. Harmful Algal Research and Response: A National Environmental Science Strategy 2005–2015”. Modeling the dynamics of an individual species is the first step towards integrating quantitative species-specific biological information into ecosystem models that take into account communities of organisms and environmental forcing. Project results are disseminated through NCCOS reports, national and international conferences, and peer reviewed publications. This project is scheduled for completion in the ten year timeframe of HARRNESS. The project is coordinated and conducted primarily at CCEHBR.