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Activation tagging using the En-I maize transposon system in Arabidopsis

Marsch-Martinez, N., Greco, R., Arkel, G. van., Herrera-Estrella, L., Pereira, A.
Plant physiology 2002 v.129 no.4 pp. 1544-1556
Arabidopsis thaliana, genetic transformation, Zea mays, food crops, insertional mutagenesis, transcriptional activation, transgenic plants, transfer DNA, corn, mutants, plant morphology, transposons
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.