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February 25, 2008
Source:
Genome Biology via
EurekAlert!
The pea is one of many important
crop species that is unsuited to the Agrobacterium-based genetic
modification techniques that are commonly used to work with
crops. Researchers, reporting in the open access journal Genome
Biology have now discovered the first high-throughput forward
and reverse genetics tool for the pea (Pisum sativum), could
have major benefits for crop breeders around the world..
Researchers from the INRA Plant Genomics Research Unit at Evry,
and the INRA Grain Legumes Research Unit at Bretenières, both in
France, both in France developed a high-quality genetic
reference collection of Pisum sativum mutants within the
European Grain Legumes Integrated Project. Abdelhafid Bendahmane
and colleagues used plants from an early-flowering garden pea
cultivar, Caméor, to create a mutant population, which they then
systematically phenotyped for use in both forward and reverse
genetics studies.
The team set up a pea TILLING (Targeting Induced Local Lesions
IN Genomes) platform with DNA samples from 4,704} plants. The
TILLING technique overcomes the pea’s natural unsuitability to
genetic modification techniques, and provides a powerful tool
for investigating the role of essential genes. This new tool has
implications for both basic science and for crop improvement.
TILLING is an alternative to Agrobacterium-based techniques, and
uses EMS (ethane methyl sulfonate) mutagenesis coupled with a
gene-specific detection of single-nucleotide mutations. This
reverse genetic strategy can be applied to all types of
organisms and can be automated for high-throughput approaches.
Following this study, the researchers created a database called
UTILLdb, which described each mutant plant at different
developmental stages, (from seedling through to fruit
maturation), and also incorporates digital images of the plants.
UTILLdb contains phenotypic as well as sequence information on
mutant genes, and can be searched for TILLING alleles of genes
of interest, using the ‘BLAST’ tool, and for plant traits of
interest, using keyword searches.
“By opening UTILLdb to the community, we hope to fulfil the
expectations of both crop breeders and scientists who are using
the pea as their model of study,” said research coordinator
Abdelhafid Bendahmane.
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UTILLdb, a Pisum sativum in silico forward and
reverse genetics tool |
Marion Dalmais, Julien Schmidt, Christine Le
Signor, Francoise Moussy, Judith Burstin, Vincent
Savois, Gregoire Aubert, Veronique Brunaud, Yannick
de Oliveira, Cecile Guichard, Richard Thompson and
Abdelhafid Bendahmane
Genome Biology
(in press)
Article available at the journal website:
http://genomebiology.com/
ABSTRACT
(provisional)
The systematic
characterisation of gene functions in species
recalcitrant to Agrobacterium-based
transformation, like Pisum sativum, remains a
challenge. To develop a high throughput forward
and reverse genetics tool in pea, we have
constructed a reference EMS-mutant population
and developed a database, UTILLdb, which
contains phenotypic as well as sequence
information on mutant genes. UTILLdb can be
searched online for TILLING alleles, through the
BLAST tool, or for phenotypic information about
mutants by keywords.
The complete
article is available as a
provisional PDF. |
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