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Flexible Tools for Gene Expression and Silencing in Tomato

Ana Fernandez Salina UGent, Nicolas Viron, Moftah Alhagdow, Mansour Karimi UGent, Matthew Jones, Ziva Amsellem, Adrien Sicard, Anna Czerednik, Gerco Angenent and Donald Grierson, et al. (2009) PLANT PHYSIOLOGY. 151(4). p.1729-1740
abstract
As a genetic platform, tomato (Solanum lycopersicum) benefits from rich germplasm collections and ease of cultivation and transformation that enable the analysis of biological processes impossible to investigate in other model species. To facilitate the assembly of an open genetic toolbox designed to study Solanaceae, we initiated a joint collection of publicly available gene manipulation tools. We focused on the characterization of promoters expressed at defined time windows during fruit development, for the regulated expression or silencing of genes of interest. Five promoter sequences were captured as entry clones compatible with the versatile MultiSite Gateway format: PPC2, PG, TPRP, and IMA from tomato and CRC from Arabidopsis (Arabidopsis thaliana). Corresponding transcriptional fusions were made with the GUS gene, a nuclear-localized GUS-GFP reporter, and the chimeric LhG4 transcription factor. The activity of the promoters during fruit development and in fruit tissues was confirmed in transgenic tomato lines. Novel Gateway destination vectors were generated for the transcription of artificial microRNA (amiRNA) precursors and hairpin RNAs under the control of these promoters, with schemes only involving Gateway BP and LR Clonase reactions. Efficient silencing of the endogenous phytoene desaturase gene was demonstrated in transgenic tomato lines producing a matching amiRNA under the cauliflower mosaic virus 35S or PPC2 promoter. Lastly, taking advantage of the pOP/LhG4 two-component system, we found that well-characterized flower-specific Arabidopsis promoters drive the expression of reporters in patterns generally compatible with heterologous expression. Tomato lines and plasmids will be distributed through a new Nottingham Arabidopsis Stock Centre service unit dedicated to Solanaceae resources.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
PLANTS, POLYGALACTURONASE, ARTIFICIAL MICRORNAS, ZINC-FINGER, FUNCTIONAL-ANALYSIS, SEQUENCE TAGS, CRABS-CLAW, ARABIDOPSIS, FRUIT, TRANSCRIPTION
journal title
PLANT PHYSIOLOGY
Plant Physiol.
volume
151
issue
4
pages
12 pages
publisher
AMER SOC PLANT BIOLOGISTS
place of publication
ROCKVILLE
Web of Science type
Article
Web of Science id
000272443500003
JCR category
PLANT SCIENCES
JCR impact factor
6.235 (2009)
JCR rank
7/172 (2009)
JCR quartile
1 (2009)
ISSN
0032-0889
DOI
10.1104/pp.109.147546
language
English
UGent publication?
yes
classification
A1
copyright statement
I don't know the status of the copyright for this publication
id
816250
handle
http://hdl.handle.net/1854/LU-816250
alternative location
http://www.plantphysiol.org/cgi/content/abstract/151/4/1729
date created
2009-12-24 17:36:33
date last changed
2010-01-04 09:42:23
@article{816250,
  abstract     = {As a genetic platform, tomato (Solanum lycopersicum) benefits from rich germplasm collections and ease of cultivation and transformation that enable the analysis of biological processes impossible to investigate in other model species. To facilitate the assembly of an open genetic toolbox designed to study Solanaceae, we initiated a joint collection of publicly available gene manipulation tools. We focused on the characterization of promoters expressed at defined time windows during fruit development, for the regulated expression or silencing of genes of interest. Five promoter sequences were captured as entry clones compatible with the versatile MultiSite Gateway format: PPC2, PG, TPRP, and IMA from tomato and CRC from Arabidopsis (Arabidopsis thaliana). Corresponding transcriptional fusions were made with the GUS gene, a nuclear-localized GUS-GFP reporter, and the chimeric LhG4 transcription factor. The activity of the promoters during fruit development and in fruit tissues was confirmed in transgenic tomato lines. Novel Gateway destination vectors were generated for the transcription of artificial microRNA (amiRNA) precursors and hairpin RNAs under the control of these promoters, with schemes only involving Gateway BP and LR Clonase reactions. Efficient silencing of the endogenous phytoene desaturase gene was demonstrated in transgenic tomato lines producing a matching amiRNA under the cauliflower mosaic virus 35S or PPC2 promoter. Lastly, taking advantage of the pOP/LhG4 two-component system, we found that well-characterized flower-specific Arabidopsis promoters drive the expression of reporters in patterns generally compatible with heterologous expression. Tomato lines and plasmids will be distributed through a new Nottingham Arabidopsis Stock Centre service unit dedicated to Solanaceae resources.},
  author       = {Fernandez Salina, Ana and Viron, Nicolas and Alhagdow, Moftah and Karimi, Mansour and Jones, Matthew and Amsellem, Ziva and Sicard, Adrien and Czerednik, Anna and Angenent, Gerco and Grierson, Donald and May, Sean and Seymour, Graham and Eshed, Yuval and Lemaire-Chamley, Martine and Rothan, Christophe and Hilson, Pierre},
  issn         = {0032-0889},
  journal      = {PLANT PHYSIOLOGY},
  keyword      = {PLANTS,POLYGALACTURONASE,ARTIFICIAL MICRORNAS,ZINC-FINGER,FUNCTIONAL-ANALYSIS,SEQUENCE TAGS,CRABS-CLAW,ARABIDOPSIS,FRUIT,TRANSCRIPTION},
  language     = {eng},
  number       = {4},
  pages        = {1729--1740},
  publisher    = {AMER SOC PLANT BIOLOGISTS},
  title        = {Flexible Tools for Gene Expression and Silencing in Tomato},
  url          = {http://dx.doi.org/10.1104/pp.109.147546},
  volume       = {151},
  year         = {2009},
}

Chicago
Fernandez Salina, Ana, Nicolas Viron, Moftah Alhagdow, Mansour Karimi, Matthew Jones, Ziva Amsellem, Adrien Sicard, et al. 2009. “Flexible Tools for Gene Expression and Silencing in Tomato.” Plant Physiology 151 (4): 1729–1740.
APA
Fernandez Salina, A., Viron, N., Alhagdow, M., Karimi, M., Jones, M., Amsellem, Z., Sicard, A., et al. (2009). Flexible Tools for Gene Expression and Silencing in Tomato. PLANT PHYSIOLOGY, 151(4), 1729–1740.
Vancouver
1.
Fernandez Salina A, Viron N, Alhagdow M, Karimi M, Jones M, Amsellem Z, et al. Flexible Tools for Gene Expression and Silencing in Tomato. PLANT PHYSIOLOGY. ROCKVILLE: AMER SOC PLANT BIOLOGISTS; 2009;151(4):1729–40.
MLA
Fernandez Salina, Ana, Nicolas Viron, Moftah Alhagdow, et al. “Flexible Tools for Gene Expression and Silencing in Tomato.” PLANT PHYSIOLOGY 151.4 (2009): 1729–1740. Print.