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Gene silencing in Fucus embryos: developmental consequences of RNAi-mediated cytoskeletal disruption

Gene silencing in Fucus embryos: developmental consequences of RNAi-mediated cytoskeletal disruption
Gene silencing in Fucus embryos: developmental consequences of RNAi-mediated cytoskeletal disruption
Brown algae (Phaeophyceae) 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 RNAi-mediated gene knockdown. We show that injection of double-stranded RNA (dsRNA) corresponding to an ?-tubulin gene into Fucus 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 dsRNA targeting ?-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 Fucus embryo possesses a functional RNA interference system that can be exploited to investigate gene function during embryogenesis.
actin, cytoskeleton, embryo, fucus, RNAi, tubulin
1529-8817
819-829
Farnham, Garry
f38067a4-8d13-4183-aa55-541ebc14aee9
Strittmatter, Martina
f0bc9d45-b2c7-44a4-8f2b-54f7fffd0ce1
Coelho, Susana
1103f9e3-7e59-467e-8e59-d7b46ff0a302
Cock, Jeremy Mark
eabaadd2-929d-419b-a548-c7138e969dc2
Brownlee, Colin
2af37c1c-b2bf-4832-8370-d9c35e7b3385
Farnham, Garry
f38067a4-8d13-4183-aa55-541ebc14aee9
Strittmatter, Martina
f0bc9d45-b2c7-44a4-8f2b-54f7fffd0ce1
Coelho, Susana
1103f9e3-7e59-467e-8e59-d7b46ff0a302
Cock, Jeremy Mark
eabaadd2-929d-419b-a548-c7138e969dc2
Brownlee, Colin
2af37c1c-b2bf-4832-8370-d9c35e7b3385

Farnham, Garry, Strittmatter, Martina, Coelho, Susana, Cock, Jeremy Mark and Brownlee, Colin (2013) Gene silencing in Fucus embryos: developmental consequences of RNAi-mediated cytoskeletal disruption. Journal of Phycology, 49 (5), 819-829. (doi:10.1111/jpy.12096).

Record type: Article

Abstract

Brown algae (Phaeophyceae) 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 RNAi-mediated gene knockdown. We show that injection of double-stranded RNA (dsRNA) corresponding to an ?-tubulin gene into Fucus 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 dsRNA targeting ?-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 Fucus embryo possesses a functional RNA interference system that can be exploited to investigate gene function during embryogenesis.

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More information

Published date: October 2013
Keywords: actin, cytoskeleton, embryo, fucus, RNAi, tubulin
Organisations: Ocean and Earth Science

Identifiers

Local EPrints ID: 361125
URI: http://eprints.soton.ac.uk/id/eprint/361125
ISSN: 1529-8817
PURE UUID: 8bf2cd3c-841b-4368-ad7f-771e9d3e2dd8

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Date deposited: 13 Jan 2014 16:50
Last modified: 09 Jan 2022 02:18

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Contributors

Author: Garry Farnham
Author: Martina Strittmatter
Author: Susana Coelho
Author: Jeremy Mark Cock
Author: Colin Brownlee

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