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Published on March 9, 2007; 10.1104/pp.106.090365

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Received September 26, 2006
Accepted January 30, 2007

Successful Gene Tagging in Lettuce (Lactuca sativa) using the Tnt1 Retrotransposon from Tobacco

Marianne Mazier , Emmanuel Botton , Fabrice Flamain , Jean-Paul Bouchet , Béatrice Courtial , Marie-Christine Chupeau , Yves Chupeau , Brigitte Maisonneuve , and Hélène Lucas *

Unité de Génétique et d'Amélioration des Fruits et Légumes, UR1502, INRA, Domaine St Maurice, BP94, F-84143 Montfavet Cedex, France; Laboratoire de Biologie Cellulaire, UR501, INRA, Route de St Cyr, F-78026 Versailles Cedex, France

* Corresponding author; email: mazier{at}avignon.inra.fr.

The tobacco element Tnt1 is one of the few identified active retrotransposons in plants. These elements possess unique properties that make them ideal genetic tools for gene tagging. Here we demonstrate the feasibility of gene tagging using the retrotransposon Tnt1 in lettuce, which is the largest genome tested for retrotransposon mutagenesis so far. Out of 10 different transgenic bushes carrying a complete Tnt1 containing T-DNA, eight contained multiple transposed copies of Tnt1. The number of transposed copies of the element per plant was particularly high, the smallest number being 28. Tnt1 transposition in lettuce can be induced by a very simple in vitro culture protocol. Tnt1 insertions were stable in the progeny of the primary transformants and could be segregated genetically. Characterization of the sequences flanking some insertion sites revealed that Tnt1 often inserted into genes. The progeny of some primary transformants showed phenotypic alterations due to recessive mutations. One of these mutations was due to Tnt1 insertion in the gibberellin 3{beta}-hydroxylase gene. Taken together, these results indicate that Tnt1 is a powerful tool for insertion mutagenesis especially in plants with a large genome.






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