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The WiscDsLox T-DNA collection: an arabidopsis community resource generated by using an improved high-throughput T-DNA sequencing pipeline

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Abstract

We have developed a new community resource, called the WiscDsLox collection, for performing reverse-genetic analysis in arabidopsis. This resource is composed of 10,459 T-DNA lines generated using the Arabidopsis thaliana ecotype Columbia. The flanking sequence tag for each T-DNA insertion has been deposited in public databases, and seed for each line is currently available from the Arabidopsis Biological Resource Center. The pDsLox vector used to create this new population contains a Ds transposon and Cre/Lox recombination sites. Each WiscDsLox line therefore has the potential to serve as a launch-pad for performing local saturation mutagenesis by mobilization of the Ds element. In addition, Cre-Lox recombination between the T-DNA and a transposed Ds element should enable targeted deletion of specific genomic regions. We generated the WiscDsLox collection using an improved high-throughput pipeline that streamlines analysis of large numbers of independent Arabidopsis thaliana (L.) Hyenh. lines. In this paper we describe the details of this novel method and also provide potential users of WiscDsLox T-DNA lines with useful background information about this collection. Experiments to characterize the utility of the Ds transposon and Cre/Lox elements present in the WiscDsLox lines are in progress and will be reported in the future.

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Acknowledgments

We acknowledge the valuable contributions of John Holt and Santoosh Raman in developing informatics resources; Sean Monson, Pete Jester, Kelly Buono, Ben Jenkins, and the many undergraduate researchers that grew and harvested WiscDsLox lines; and Pam Ziegelhoffer, Arabidopsis Knockout facility, for managing line distribution. Special thanks are extended to Huaming Chen and the Ecker group at the Salk Institute for integration of WiscDsLox lines via T-DNA Express and to Chris Day for useful comments on the manuscript. This work was supported by the National Science Foundation Arabidopsis 2010 Project No. 0116945 and National Science Foundation Arabidopsis 2010 Project No. DBI-0514171.

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Authors and Affiliations

  1. Horticulture Department and Genome Center of Wisconsin, University of Wisconsin-Madison, Madison, WI, 53706, USA

    Scott T. Woody & Patrick J. Krysan

  2. Biotechnology Center, University of Wisconsin-Madison, Madison, WI, 53706, USA

    Scott T. Woody & Sandra Austin-Phillips

  3. Biochemistry Department, University of Wisconsin-Madison, Madison, WI, 53706, USA

    Richard M. Amasino

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  1. Scott T. Woody
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  2. Sandra Austin-Phillips
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  3. Richard M. Amasino
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  4. Patrick J. Krysan
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Correspondence to Patrick J. Krysan.

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Woody, S.T., Austin-Phillips, S., Amasino, R.M. et al. The WiscDsLox T-DNA collection: an arabidopsis community resource generated by using an improved high-throughput T-DNA sequencing pipeline. J Plant Res 120, 157–165 (2007). https://doi.org/10.1007/s10265-006-0048-x

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  • DOI: https://doi.org/10.1007/s10265-006-0048-x

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