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Gene silencing in F ucus embryos: developmental consequences of RNA i‐mediated cytoskeletal disruption
Author(s) -
Farnham Garry,
Strittmatter Martina,
Coelho Susana,
Cock Jeremy Mark,
Brownlee Colin
Publication year - 2013
Publication title -
journal of phycology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.85
H-Index - 127
eISSN - 1529-8817
pISSN - 0022-3646
DOI - 10.1111/jpy.12096
Subject(s) - biology , fucales , rna interference , microbiology and biotechnology , tip growth , gene knockdown , gene expression , gene silencing , rhizoid , rna , gene , rna silencing , actin , cytoskeleton , cell polarity , microtubule , genetics , cell , algae , botany , pollen , pollination , pollen tube
Brown algae ( P haeophyceae) are an important algal class that play a range of key ecological roles. They are often important components of rocky shore communities. A number of members of the Fucales and Ectocarpales have provided models for the study of multicellular evolution, reproductive biology and polarized development. Indeed the fucoid algae exhibit the unusual feature of inducible embryo polarization, allowing many classical studies of polarity induction. The potential of further studies of brown algae in these important areas has been increasingly hindered by the absence of tools for manipulation of gene expression that would facilitate further mechanistic analysis and gene function studies at a molecular level. The aim of this study was to establish a method that would allow the analysis of gene function through RNA i‐mediated gene knockdown. We show that injection of double‐stranded RNA (ds RNA ) corresponding to an α‐tubulin gene into F ucus serratus Linnaeus zygotes induces the loss of a large proportion of the microtubule cytoskeleton, leading to growth arrest and disruption of cell division. Injection of ds RNA targeting β‐actin led to reduced rhizoid growth, enlarged cells and the failure to develop apical hair cells. The silencing effect on actin expression was maintained for 3 months. These results indicate that the F ucus embryo possesses a functional RNA interference system that can be exploited to investigate gene function during embryogenesis.