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Published on July 18, 2002; 10.1104/pp.003327

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Received January 29, 2002
Returned for revision February 26, 2002
Accepted April 8, 2002

Activation Tagging Using the En-I Maize Transposon System in Arabidopsis

Nayelli Marsch-Martinez , Raffaella Greco , Gert Van Arkel , Luis Herrera-Estrella , and Andy Pereira *

Plant Research International, P.O. Box 16, 6700 AA Wageningen, The Netherlands (N.M.-M., R.G., G.V.A., A.P.); and Centro de Investigación y de Estudios Avanzados-Instituto Politécnico Nacional-Irapuato, P.O. Box 629, 3500 Irapuato, Guanajuato, México (N.M.-M., L.H.-E.)

* Corresponding author; email: A.Pereira{at}plant.wag-ur.nl.

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.




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