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Nucleic Acids Research Advance Access originally published online on May 31, 2009
Nucleic Acids Research 2009 37(14):e96; doi:10.1093/nar/gkp448
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Nucleic Acids Research, 2009, Vol. 37, No. 14 e96
© 2009 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Gene silencing in the marine diatom Phaeodactylum tricornutum

Valentina De Riso1, Raffaella Raniello1, Florian Maumus2, Alessandra Rogato1, Chris Bowler2,3 and Angela Falciatore1,*

1Laboratory of Ecology and Evolution of Plankton, Stazione Zoologica Anton Dohrn, Villa Comunale, 80121, Napoli, Italy, 2CNRS UMR 8186, Department of Biology, Ecole Normale Supérieure, 46 rue d'Ulm, 75005, Paris, France and 3Stazione Zoologica Anton Dohrn, Villa Comunale, 80121, Napoli, Italy

*To whom correspondence should be addressed. Tel: +39 081 5833268; Fax: +39 081 7641355; Email: afalciat{at}szn.it

Received November 11, 2008. Revised May 6, 2009. Accepted May 12, 2009.

Diatoms are a major but poorly understood phytoplankton group. The recent completion of two whole genome sequences has revealed that they contain unique combinations of genes, likely recruited during their history as secondary endosymbionts, as well as by horizontal gene transfer from bacteria. A major limitation for the study of diatom biology and gene function is the lack of tools to generate targeted gene knockout or knockdown mutants. In this work, we have assessed the possibility of triggering gene silencing in Phaeodactylum tricornutum using constructs containing either anti-sense or inverted repeat sequences of selected target genes. We report the successful silencing of a GUS reporter gene expressed in transgenic lines, as well as the knockdown of endogenous phytochrome (DPH1) and cryptochrome (CPF1) genes. To highlight the utility of the approach we also report the first phenotypic characterization of a diatom mutant (cpf1). Our data open the way for reverse genetics in diatoms and represent a major advance for understanding their biology and ecology. Initial molecular analyses reveal that targeted downregulation likely occurs through transcriptional and post-transcriptional gene silencing mechanisms. Interestingly, molecular players involved in RNA silencing in other eukaryotes are only poorly conserved in diatoms.


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