Rapid inactivation of the maize transposable element En/Spm in Medicago truncatula.
Transposable elements have been widely used as mutagens in many organisms. Among them, the maize transposable element En/Spm has been shown to transpose efficiently in several plant species including the model plant Arabidopsis, where it has been used for large-scale mutagenesis. To determine whether we could use this transposon as a mutagen in the model legume plant Medicago truncatula, we tested the activity of the autonomous element, as well as two defective elements, in this plant, and in Arabidopsis as a positive control. In agreement with previous reports, we observed efficient excision of the autonomous En/Spm element in A. thaliana. This element was also active in M. truncatula, but the transposition activity was low and was apparently restricted to the tissue culture step necessary for the production of transgenic plants. The activity of one of the defective transposable elements, dSpm, was very low in A. thaliana and even lower in M. truncatula. The use of different sources of transposases suggested that this defect in transposition was associated with the dSpm element itself. Transposition of the other defective element, I6078, was also detected in M. truncatula, but, as observed with the autonomous element, transposition events were very rare and occurred during tissue culture. These results suggest that the En/Spm element is rapidly inactivated in the regenerated plants and their progeny, and therefore is not suitable for routine insertion mutagenesis in M. truncatula.
d'Erfurth
I
I
Institut des Sciences du Végétal, CNRS, Avenue de la Terrasse, 91198 Gif sur Yvette, France.
Cosson
V
V
Eschstruth
A
A
Rippa
S
S
Messinese
E
E
Durand
P
P
Trinh
H
H
Kondorosi
A
A
Ratet
P
P
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D017931:DNA Primers / Q000737:chemistry; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D007612:Kanamycin / Q000494:pharmacology; D029902:Medicago / Q000235:genetics* / Q000254:growth & development; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010641:Phenotype; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D011995:Recombination, Genetic; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D014170:Transformation, Genetic; D019895:Transposases / Q000378:metabolism; D003313:Zea mays / Q000235:genetics*
D017931:DNA Primers; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D018744:DNA, Plant; D012333:RNA, Messenger; C036483:T-DNA; D007612:Kanamycin; D019895:Transposases; D005966:Glucuronidase