Efficient insertional mutagenesis in rice using the maize En/Spm elements

doi:10.1111/j.1365-313X.2005.02570.x

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Wiley InterScience

The Plant Journal

Volume 44 Issue 5, Pages 879 - 892

Published Online: 21 Oct 2005

Published in association with the Society for Experimental Biology

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TECHNICAL ADVANCE

Efficient insertional mutagenesis in rice using the maize En/Spm elements

Chellian Santhosh Kumar ¹ , Rod A. Wing ² and Venkatesan Sundaresan ^1,3,*

¹ Department of Plant Sciences, Life Sciences Addition 1002, University of California, One Shields Ave, Davis, CA 95616, USA,
² Arizona Genomics Institute, 303 Forbes Building, University of Arizona, Tucson, AZ 85721, USA, and
³ Section of Plant Biology, Life Sciences Addition 1002, University of California, One Shields Ave, Davis, CA 95616 USA

Correspondence to *(fax +1 530 752 5410; e-mail sundar@ucdavis.edu).

KEYWORDS

Oryza sativa • En/Spm-I/dSpm • transposon tagging • unlinked transposition • dSpm flanking sequence tags • functional genomics

Summary

We have developed a novel system for insertional mutagenesis in rice (Oryza sativa) based on the maize (Zea mays) enhancer/suppressor mutator (En/Spm) element. In this system, a single T-DNA construct with Spm-transposase and the non-autonomous defective suppressor mutator (dSpm) element is used in conjunction with green fluorescent protein (GFP) and Discosoma sp. Red Fluorescence Protein (DsRed) fluorescent markers to select unlinked stable transpositions of dSpm. Using this system, we could demonstrate high frequencies of unlinked germinal transposition of dSpm in rice. Analysis of dSpm flanking sequences from 353 stable insertion lines revealed that the dSpm insertions appear to be widely distributed on rice chromosomes with a preference for genic regions (70%). The dSpm insertions appear to differ from Activator-Dissociation (Ac-Ds) elements in genomic distribution and exhibit a greater fraction of unlinked transpositions when compared with Ds elements. The results obtained in this study demonstrate that the maize En/Spm element can be used as an effective tool for functional genomics in rice and can complement efforts using other insertional mutagens. Further, the efficacy of the non-invasive fluorescence-based selection system is promising for its application to other crops.