Abstract
Diatoms are single-celled eukaryotic microalgae that are ubiquitously found in almost all aquatic ecosystems, and are characterized by their intricately structured SiO2 (silica)-based cell walls. Diatoms with a benthic life style are capable of attaching to any natural or man-made submerged surface, thus contributing substantially to both microbial biofilm communities and economic losses through biofouling. Surface attachment of diatoms is mediated by a carbohydrate- and protein- based glue, yet no protein involved in diatom underwater adhesion has been identified so far. In the present work, we have generated a normalized transcriptome database from the model adhesion diatom Amphora coffeaeformis. Using an unconventional bioinformatics analysis we have identified five proteins that exhibit unique amino acid sequences resembling the amino acid composition of the tyrosine-rich adhesion proteins from mussel footpads. Establishing the first method for the molecular genetic transformation of A. coffeaeformis has enabled investigations into the function of one of these proteins, AC3362, through expression as YFP fusion protein. Biochemical analysis and imaging by fluorescence microscopy revealed that AC3362 is not involved in adhesion, but rather plays a role in biosynthesis and/or structural stability of the cell wall. The methods established in the present study have paved the way for further molecular studies on the mechanisms of underwater adhesion and biological silica formation in the diatom A. coffeaeformis.
Highlights
Diatoms are a large group of single-celled microalgae that are ubiquitously present in water habitats, and are among the most prolific biological primary producers in the oceans [1]
AC3362 was identified by screening a normalized A. coffeaeformis transcriptome database, which has been established in the present study
Studies on the functional characterization of AC3362 relied on a genetic transformation system for A. coffeaeformis that has been established in the present study
Summary
Diatoms are a large group of single-celled microalgae that are ubiquitously present in water habitats, and are among the most prolific biological primary producers in the oceans [1]. Diatoms are widely studied due to their importance for ocean ecosystems, their physiological capabilities, their complex evolutionary history, and their ability to adhere to any natural or man-made surface underwater. The colonization of submerged surfaces (‘‘biofouling’’) by bacteria, microalgae (including diatoms), and multicellular organisms (e.g., barnacles, mussels and macroalgae) can lead to the development of biofilms that are several centimeters thick [2]. The adhesive components produced by diatoms provide a paradigm for the development of underwater glues for numerous applications in technology and medicine [7]. The prevention of diatom adhesion and the development of underwater glues, it is necessary to identify the adhesive biomolecules of diatoms and understand their molecular mechanism of adhesion
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