Activation tagging using the En-I maize transposon system in Arabidopsis.
A method for the generation of stable activation tag inserts was developed in Arabidopsis using the maize (Zea mays) En-I transposon system. The method employs greenhouse selectable marker genes that are useful to efficiently generate large populations of insertions. A population of about 8,300 independent stable activation tag inserts has been produced. Greenhouse-based screens for mutants in a group of plants containing about 2,900 insertions revealed about 31 dominant mutants, suggesting a dominant mutant frequency of about 1%. From the first batch of about 400 stable insertions screened in the greenhouse, four gain-in-function, dominant activation-tagged, morphological mutants were identified. A novel gain-in-function mutant called thread is described, in which the target gene belongs to the same family as the YUCCA flavin-mono-oxygenase that was identified by T-DNA activation tagging. The high frequency of identified gain-in-function mutants in the population suggests that the En-I system described here is an efficient strategy to saturate plant genomes with activation tag inserts. Because only a small number of primary transformants are required to generate an activation tag population, the En-I system appears to be an attractive alternative to study plant species where the present transformation methods have low efficiencies.
Marsch-Martinez
Nayelli
N
Plant Research International, P.O. Box 16, 6700 AA Wageningen, The Netherlands.
Greco
Raffaella
R
Van Arkel
Gert
G
Herrera-Estrella
Luis
L
Pereira
Andy
A
D024509:3' Flanking Region / Q000235:genetics; D024506:5' Flanking Region / Q000235:genetics; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins; D004251:DNA Transposable Elements / Q000235:genetics*; D020869:Gene Expression Profiling; D016254:Mutagenesis, Insertional / Q000379:methods; D009154:Mutation; D010105:Oxygenases / Q000235:genetics; D030821:Plants, Genetically Modified; D019895:Transposases / Q000378:metabolism; D003313:Zea mays / Q000235:genetics*
D029681:Arabidopsis Proteins; D004251:DNA Transposable Elements; D010105:Oxygenases; C422746:YUC protein, Arabidopsis; D019895:Transposases