T-DNA tagging in the model legume Medicago truncatula allows efficient gene discovery

The annual legume Medicago truncatula has been proposed as a model plant to study various aspects of legume biology including rhizobial and mycorrhizal symbiosis because it is well suited for the genetic analysis of these processes . To facilitate the characterization of M. truncatula genes particip...

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Bibliographic Details
Published in:Molecular breeding 2002-01, Vol.10 (4), p.203-215
Main Authors: Scholte, Marije, d'Erfurth, Isabelle, Rippa, Sonia, Mondy, Samuel, Cosson, Viviane, Durand, Patricia, Breda, Colette, Trinh, Hanh, Rodriguez-Llorente, Ignacio, Kondorosi, Eva, Schultze, Michael, Kondorosi, Adam, Ratet, Pascal
Format: Article
Language:English
Subjects:
Alfalfa
Copy number
Deoxyribonucleic acid
DNA
DNA structure
Gene sequencing
Genes
Genetic analysis
Insertion
Inserts
Kanamycin
Legumes
Life Sciences
Marking
Medicago truncatula
Molecular biology
Mutagenesis
Nucleotide sequence
Plant biology
Symbiosis
T-DNA
Transgenic plants
Vegetal Biology
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Summary:The annual legume Medicago truncatula has been proposed as a model plant to study various aspects of legume biology including rhizobial and mycorrhizal symbiosis because it is well suited for the genetic analysis of these processes . To facilitate the characterization of M. truncatula genes participating in various developmental processes we have initiated an insertion mutagenesis program in this plant using three different T-DNAs as tags. To investigate which type of vector is the most suitable for mutagenesis we compared the behavior of these T-DNAs. One T-DNA vector was a derivative of pBin19 and plant selection was based on kanamycin resistance. The two other vectors carried T-DNA conferring Basta resistance in the transgenic plants. For each T-DNA type, we determined the copy number in the transgenic lines, the structure of the T-DNA loci and the sequences of the integration sites. The T-DNA derived from pBin19 generated complex T-DNA insertion patterns. The two others generally gave single copy T-DNA inserts that could result in gene fusions for the pGKB5 T-DNA. Analysis of the T-DNA borders revealed that several M. truncatula genes were tagged in these transgenic lines and in vivo gus fusions were also obtained. These results demonstrate that T-DNA tagging can efficiently be used in M. truncatula for gene discovery.
ISSN:1380-3743
1572-9788
DOI:10.1023/A:1020564612093