Project Goal

Targeted mutagenesis with the maize transposable element Ds in barley is made possible by a tendency for the element to reinsert near the original site of excision. The aim of this project is the generation of populations of individual barley lines, each with an independently transposed, mapped Ds element, such that each region of the barley genome (bin) is represented by at least one line containing a Ds insertion. To exploit localized transposition for gene tagging, transgenic barley stocks with insertions located near genes of interest will be candidates for secondary activation of the elements to permit efficient tagging based on short-range transposition of Ds.

Project Results

January 2006: To date 137 lines with transposed Ds-bar elements have been identified in barley. Of those 137, flanking sequences have been determined for ~80 lines and currently 35 have been mapped (19 map locations were published (see below); Cooper et al., 2004.

Publications:

• Cooper LD, Marquez-Cedillo L, Singh J, Sturbaum AK, Zhang S, Edwards V, Johnson K, Kleinhofs A, Rangel S, Carollo V, Bregitzer P, Lemaux PG, Hayes PM (2004) Mapping Ds insertions in barley using a sequence based approach. Molecular Genetics and Genomics 272: 181-193. (Pubmed Link)

• Singh J, Zhang S, Lemaux PG, Bregitzer P, Sturbaum AK, Edwards V, Hayes PM, Cooper LD, Marquez-Cedillo L, Carollo V (2005) Maize Transposable Elements and Barley: A New Population for Genetic Research. Barley Genetics Newsletter 35: 1-2

• Zhang S, Chen C, Lei L, Meng L, Singh J, Jiang N, Deng X-W, He Z-H, Lemaux PG (2005) Evolutionary Expansion, Gene Structure, and Expression of the Rice Wall-Associated Kinase Gene Family. Plant Physiology 139:1107-1124. (Pubmed Link)

• Singh J, Zhang S, Chen C, Cooper L, Bregitzer P, Sturbaum AK, Hayes PM, Lemaux PG (2006) High-frequency Reactivation of Maize Ds Favors Saturation Mutagenesis in Barley. Plant Molecular Biology DOI 10.1007/s11103-006-9067-1.

• Meng L, Ziv M, Lemaux PG (2006) Nature of Stress and Transgene Locus Influences Transgene Expression Stability in Barley. Plant Molecular Biology 62: 15-28 Pubmed link

• Bregitzer, P., L. Cooper, P. Hayes, P.G. Lemaux, J. Singh, and A. Sturbaum. (2006) Viability and bar expression are negatively correlated in Oregon Wolfe Barley dominant hybrids. Plant Biotechnology Journal PubMed

• Randhawa, P.G., Singh, J., H. Lemaux, and K.S. Gill. (2009) Mapping barley Ds insertions using wheat deletion lines reveals high insertion frequencies in gene-rich regions with high to moderate recombination rates. Genome PubMed

The Interactive Oregon Wolfe Barley Maps (IMap) provide an interactive resource to view barley bin maps with links to information on the individual TNP lines, naked-eye phenotypes (NEPs) and provides links to the GrainGenes database from all other loci.

Lemaux Lab, University of California, Berkeley

Peggy G. Lemaux   (lemauxpg@nature.Berkeley.edu)

Jaswinder Singh

Shibo Zhang

Hayes Lab, Oregon State University

Patrick  Hayes 

Lol Cooper

Luis Marquez-Cedillo

Bregitzer Lab, National Small Grains Germplasm Research Facility, USDA-ARS, Aberdeen, Idaho

Phil Bregitzer (Phil.Bregitzer@ars.usda.gov )

Anne Sturbaum

GrainGenes Project, USDA-ARS-WRRC, Albany, CA

Victoria Carollo Blake (vcarolloblake@gmail.com)

Instructions for Requesting TNP lines and Ac Transposase lines

APHIS Notification for Interstate Transfer form

• Word document (.doc)
  

Example of a Standard Operating Procedure (SOP) for Growing and Agronomic Testing of Transgenic Barley
(Hordeum vulgare) and Wheat (Triticum aestivum) Plants.

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• Word document (.doc)