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Published on November 9, 2007; 10.1104/pp.107.111427

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Received October 22, 2007
Accepted October 30, 2007

A Versatile Transposon-Based Activation Tag Vector System for Functional Genomics in Cereals and Other Monocot Plants

Shaohong Qu , Aparna Desai , Rod Wing , and Venkatesan Sundaresan *

Section of Plant Biology, University of California, Davis, California 95616; Department of Plant Sciences, University of California, Davis, California 95616; Arizona Genomics Institute, University of Arizona, Tucson, Arizona 85721

* Corresponding author; email: sundar{at}ucdavis.edu.

Transposon insertional mutagenesis is an effective alternative to T-DNA mutagenesis when transformation through tissue culture is inefficient as is the case for many crop species. When used as activation tags, transposons can be exploited to generate novel gain-of-function phenotypes without transformation, and are of particular value in the study of polyploid plants where gene knockouts will not have phenotypes. We have developed an in cis activation tagging Ac-Ds system in which a T-DNA vector carries a Ds element containing 4x CaMV enhancers along with the Ac transposase gene. Stable Ds insertions were selected using dual GFP/RFP fluorescence marker genes driven by promoters that are functional in maize (Zea mays) and rice (Oryza sativa). The system has been tested in rice, where 638 stable Ds insertions were selected from an initial set of 26 primary transformants. By analysis of 311 flanking sequences mapped to the rice genome, we could demonstrate the wide distribution of the elements over the rice chromosomes. Enhanced expression of rice genes adjacent to Ds insertions was detected in the insertion lines using semi-quantitative RT-PCR method. The in cis 2-element vector system requires minimal number of primary transformants and eliminates the need for crossing, while the use of fluorescent markers instead of antibiotic or herbicide resistance increases the applicability to other plants and eliminates problems with escapes. Since Ac-Ds has been shown to transpose widely in the plant kingdom, the activation vector system developed in this study should be of utility more generally to other monocots.




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