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BAC Library Development and Clone Characterization for Dormancy-Responsive DREB4A, DAM, and FT from Leafy Spurge (Euphorbia esula) Identifies Differential Splicing and Conserved Promoter Motifs

Published online by Cambridge University Press:  20 January 2017

David P. Horvath ,
David Kudrna ,
Jayson Talag ,
James V. Anderson ,
Wun S. Chao ,
Rod Wing ,
Michael E. Foley  and
Münevver Doğramacı
David P. Horvath*
Affiliation:
Sunflower and Plant Biology Research Unit, Red River Valley Agricultural Research Center, U.S. Department of Agriculture–Agricultural Research Service, Fargo, ND 58102
David Kudrna
Affiliation:
Arizona Genomics Institute, School of Plant Sciences, University of Arizona, Tucson, AZ 85721
Jayson Talag
Affiliation:
Arizona Genomics Institute, School of Plant Sciences, University of Arizona, Tucson, AZ 85721
James V. Anderson
Affiliation:
Sunflower and Plant Biology Research Unit, Red River Valley Agricultural Research Center, U.S. Department of Agriculture–Agricultural Research Service, Fargo, ND 58102
Wun S. Chao
Affiliation:
Sunflower and Plant Biology Research Unit, Red River Valley Agricultural Research Center, U.S. Department of Agriculture–Agricultural Research Service, Fargo, ND 58102
Rod Wing
Affiliation:
Arizona Genomics Institute, School of Plant Sciences, University of Arizona, Tucson, AZ 85721
Michael E. Foley
Affiliation:
Sunflower and Plant Biology Research Unit, Red River Valley Agricultural Research Center, U.S. Department of Agriculture–Agricultural Research Service, Fargo, ND 58102
Münevver Doğramacı
Affiliation:
Sunflower and Plant Biology Research Unit, Red River Valley Agricultural Research Center, U.S. Department of Agriculture–Agricultural Research Service, Fargo, ND 58102
*
Corresponding author's E-mail: [email protected]
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Abstract

We developed two leafy spurge bacterial artificial chromosome (BAC) libraries that together represent approximately 5× coverage of the leafy spurge genome. The BAC libraries have an average insert size of approximately 143 kb, and copies of the library and filters for hybridization-based screening are publicly available through the Arizona Genomics Institute. These libraries were used to clone full-length genomic copies of an AP2/ERF transcription factor of the A4 subfamily of DEHYDRATION-RESPONSIVE ELEMENT-BINDING PROTEINS (DREB) known to be differentially expressed in crown buds of leafy spurge during endodormancy, a DORMANCY ASSOCIATED MADS-BOX (DAM) gene, and several FLOWERING LOCUS T (FT) genes. Sequencing of these BAC clones revealed the presence of multiple FT genes in leafy spurge. Sequencing also provided evidence that two different DAM transcripts expressed in crown buds of leafy spurge during endo- and eco-dormancy result from alternate splicing of a single DAM gene. Sequence data from the FT promoters was used to identify several conserved elements previously recognized in Arabidopsis, as well as potential novel transcription factor binding sites that may regulate FT. These leafy spurge BAC libraries represent a new genomics-based tool that complements existing genomics resources for the study of plant growth and development in this model perennial weed. Furthermore, phylogenetic footprinting using genes identified with this resource demonstrate the usefulness of studying weedy species to further our general knowledge of agriculturally important genes.

Keywords

Arabidopsis, Arabidopsis thaliana L. ARATHleafy spurge, Euphorbia esula L. EPHESAlternate splicingfloweringphylogenetic footprintingtranscription factors
Type
Weed Biology and Ecology
Information
Weed Science , Volume 61 , Issue 2 , June 2013 , pp. 303 - 309
Copyright
Copyright © Weed Science Society of America 

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