The Arabidopsis Functional Genomics Network


AtGenExpress at databases:
Developmental samples
Stress treatments
GARNET: Light treatments, Pathogen infection, Base
line experiments
Overview
- General information: Grant proposal (new proposal), Project: AtGenExpress:
A multinational coordinated effort to uncover the transcriptome of the multicellular
model organism Arabidopsis thaliana (A project coordinated by Detlef Weigel,
Thomas Altmann and Lutz Nover)
- Principal investigator for AFGN: Prof. Dr. Lutz Nover, Botanisches Institut,
Molekulare Zellbiologie, FB Biologie und Informatik der J. W. Goethe Universität
Frankfurt; Biozentrum N200 3OG, Marie-Curie-Strasse 9, 60439 Frankfurt am
Main, nover@cellbiology.uni-frankfurt.de
- Summary
The Affymetrix technology for expression profiling with the ATH1 full genome
chip allows the reliable detection of about 24,000 protein-coding genes of
Arabidopsis. It includes the RNA processing and labelling technology and the
Gene Array Scanner with workstation and data processing software. Altogether
this represents a unique combination of tools broadly used worldwide. Based
on this technology, the AFGN groups propose an international cooperation project
(AtGenExpress) to generate a gene expression profiling database covering about
230 data sets derived from 500 RNA samples with financial support by the DFG.
This part includes samples for Arabidopsis development, stress responses,
light response and pathogen interaction. It will be complemented by 80 data
sets from the NSF 2010 project (USA) on pathogen responses, by 170 data sets
from RIKEN (Japan) on hormone responses and by about 50 data sets from the
MPIs in Tübingen and Golm. The overall data base derived from more than
1300 microarrays, i.e. more than 30 million data points, will be released
to the public as part of the international Gene Expression Omnibus (GEO) and
ArraExpress data bases. Data processing and publication will be mannaged by
the NSF-supported Arabidopsis database (TAIR, USA).
AtGenExpress: A multinational coordinated effort to uncover the transcriptome
of the multicellular model organism Arabidopsis thaliana
Coordinated by Detlef Weigel(1), Thomas Altmann(2), and Lutz Nover(3)
(1) Department of Molecular Biology, MPI for Developmental Biology, Spemannstrasse
37-39, D-72076 Tübingen
(2) Institut für Biochemie und Biologie, Universität Potsdam, c/o
MPI für molekulare Pflanzenphysiologie, Am Mühlenberg 1, D-14476
Golm
(3) Molekulare Zellbiologie, Biozentrum, Goethe Universität Frankfurt,
Marie-Curie-Str. 09, D-60439 Frankfurt
- Executive Summary
In the 1990s, the genomes of several multicellular eukaryotic model organisms
were sequenced, including those of the nematode Caenorhabditis elegans, the
fruit fly Drosophila melanogaster and the crucifer Arabidopsis thaliana. However,
DNA sequencing can only be considered as the first step in understanding the
function of these genomes. The Arabidopsis community has been at the forefront
of initiating concerted actions toward functional genomics. For example, the
ambitious goal of creating knock out lines for every Arabidopsis gene has
almost been reached through the generation of sequence-defined T-DNA and transposon
lines. Arabidopsis is also the only organism among the three aforementioned
where already considerable information on natural allelic and genetic diversity
exists, and it is the only species, apart from humans, where genome-wide linkage
disequilibrium has been measured.
In contrast to the genome sequencing of Arabidopsis with significant contributions
from Germany and other European countries, there was so far no major technological
impact on the functional genomics phase. Here, we propose a large-scale service
project for genome-wide expression profiling in Arabidopsis (AtGenExpress)
designed and coordinated by the German Arabidopsis Functional Genomics Network
(AFGN), but incorporating strong elements of international cooperation. To
our knowledge, no other comparable initiative exists. Its completion and public
presentation depends on the strong support by the DFG, but it requires very
substantial contributions by RIKEN (Japan), NSF (USA; via funding of TAIR
and the 2010 program) , BBSRC (UK; via funding of the GARNET initiative),
and the Max-Planck-Society.
The expression profiles will be determined using the Affymetrix ATH1 gene
chip for analysis of the Arabidopsis thaliana wild type (var. Col-0). The
created data sets will not only serve as the main reference to the Arabidopsis
transcriptome, but will serve as the principal information resource for data
mining by Arabidopsis researchers worldwide. This will create an unprecedented
opportunity for the establishment of new hypotheses and thus will aid researchers
to rationalize detailed studies. Most notably, the functional genomics analyses
conducted in the AFGN and 2010 projects will strongly benefit by providing
the tools to specifically design studies with mutants created by T-DNA insertions
or RNAi knock-out. Finally, AtGenExpress will be an indispensable reference
database also for research with other plants, in particular with crop plants
for which only microarrays with a limited set of gene-specific probes are
available.
We propose to generate a sufficiently comprehensive gene expression profile
covering about 230 data sets from Arabidopsis development, stress responses,
light response and pathogen interaction (Tables 1-3) complemented by the above
mentioned contributions of about 290 data sets each 20 from the MPIs in Tübingen
(D. Weigel) and Golm (M. Stitt), 80 from the NSF 2010 project (pathogen responses)
and 170 from RIKEN (hormone responses). The overall database represents an
open system encouraging the addition of further, more specific data sets.
Thus, it will continue to grow in its usefulness.
The AtGenExpress project sensu stricto has three phases (for a time schedule
see 7.):
1. Selected expert groups will generate RNA samples in triplicate or duplicate
from the plant material defined in Tables 1-3.
2. To achieve jointly the necessary throughput, three sites (MPI Tübingen,
RZPD Berlin, and the GARNET microarray facility at Nottingham) will be used
for the processing of the RNA and generation of the expression data with Affymetrix
ATH1 arrays. All have long experience in Affymetrix technology and data handling
and apply the same standardized procedures.
3. All data will be released as soon as possible to the Gene Expression Omnibus
(GEO) database of NCBI. Data for each gene will be displayed on the NSF-supported
Arabidopsis database (TAIR). At this stage, a web server will be set up that
will allow data mining of researchers from all over the world. In addition,
work stations with the Affymetrix software at different places in Germany
(Tübingen, Potsdam, Halle, Frankfurt) will provide the means for data
mining by AFGN co-workers.
Coordinator: Lutz Nover,
Detlef Weigel and Thomas
Altmann