Journal Article

Osmotic shock improves Tnt1 transposition frequency in Medicago truncatula cv Jemalong during in vitro regeneration

Ratet, Pascal, Primard-Brisset, Catherine, Barascud, Marielle, Cosson, Viviane, Chabaud, Mireille, Durand, Patricia, Iantcheva, Anelia, Unité mixte de recherche interactions plantes-microorganismes ; Institut National de la Recherche Agronomique (INRA)-Université Toulouse III - Paul Sabatier (UT3) ; Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Schutz, Bernadette, Vlahova, Mariana, European FP6 : FOOD-CT-2004-506223 GRAIN LEGUMES IP, AgroBioInstitute ; AgroBioInstitute, Barker, David, Institut des Sciences Végétales ; Centre National de la Recherche Scientifique (CNRS)

2009

Overview

Insertion mutant collections are powerful tools for genetic studies in plants. Although large-scale insertional mutagenesis using T-DNA is not feasible in legumes, the Tnt1 tobacco retrotransposon can be used as a very efficient mutagen in the Medicago truncatula R108 genotype. In this article, we show that Tnt1 can also be exploited to create insertional mutants via transformation and/or regeneration in the reference cultivar Jemalong. Tnt1 insertional mutagenesis in Jemalong following Agrobacterium tumefaciens-mediated transformation was found to be very efficient, with an average of greater than 15 insertions/line. In contrast, regeneration using low-copy transgenic starter lines resulted in a highly variable rate of new Tnt1 insertions. With the goal of increasing the number of additional Tnt1 insertions during regeneration of starter lines, we have compared the insertion frequencies for a number of different regeneration protocols. In addition, we have been able to show that sucrose-mediated osmotic shock preceding regeneration significantly increases the transposition frequency. Under optimal conditions, 95% of the regenerated Jemalong plants possess new insertions.

• Publisher: Springer Verlag,CCSD,Springer-Verlag,Springer,Springer Nature B.V
• Subject: Agrobacterium tumefaciens - genetics / Alfalfa / Biological and medical sciences / Biomedical and Life Sciences / Biotechnology / Cell Biology / Cultivars / Culture Media / DNA, Plant - genetics / Fundamental and applied biological sciences. Psychology / Gene Expression Regulation, Plant / Genetics / Life Sciences / Medicago truncatula - genetics / Medicago truncatula - metabolism / Mutagenesis, Insertional - methods / Original Paper / Osmotic Pressure / Plant Biochemistry / Plant Sciences / Plants genetics / Plants, Genetically Modified - genetics / Plants, Genetically Modified - metabolism / Regeneration / Retroelements / Sucrose - pharmacology / Transformation, Genetic
• ISBN: 0002701867000