caa a22 4500 46789504X CHVBK 20180406152800.0 cr unu---uuuuu 170328e20061201xx s 000 0 eng 10.1007/s11103-006-9067-1 doi (NATIONALLICENCE)springer-10.1007/s11103-006-9067-1 High-frequency Ds remobilization over multiple generations in barley facilitates gene tagging in large genome cereals [Elektronische Daten] [Jaswinder Singh, Shibo Zhang, Calvin Chen, Laurel Cooper, Phil Bregitzer, Anne Sturbaum, Patrick Hayes, Peggy Lemaux] Transposable elements have certain advantages over other approaches for identifying and determining gene function in large genome cereals. Different strategies have been used to exploit the maize Activator/dissociation (Ac/Ds) transposon system for functional genomics in heterologous species. Either large numbers of independent Ds insertion lines or transposants (TNPs) are generated and screened phenotypically, or smaller numbers of TNPs are produced, Ds locations mapped and remobilized for localized gene targeting. It is imperative to characterize key features of the system in order to utilize the latter strategy, which is more feasible in large genome cereals like barley and wheat. In barley, we generated greater than 100 single-copy Ds TNPs and determined remobilization frequencies of primary, secondary, and tertiary TNPs with intact terminal inverted repeats (TIRs); frequencies ranged from 11.8 to 17.1%. In 16% of TNPs that had damaged TIRs no transposition was detected among progeny of crosses using those TNPs as parental lines. In half of the greater than 100 TNP lines, the nature of flanking sequences and status of the 11bp TIRs and 8-bp direct repeats were determined. BLAST searches using a gene prediction program revealed that 86% of TNP flanking sequences matched either known or putative genes, indicating preferential Ds insertion into genic regions, critical in large genome species. Observed remobilization frequencies of primary, secondary, tertiary, and quaternary TNPs, coupled with the tendency for localized Ds transposition, validates a saturation mutagenesis approach using Ds to tag and characterize genes linked to Ds in large genome cereals like barley and wheat. Springer Science+Business Media B.V., 2006 Ac /Ds nationallicence Barley nationallicence Remobilization nationallicence Transposon tagging nationallicence Ac/Ds : Activator/dissociation nationallicence EST : Expressed sequence tag nationallicence TIR : Terminal inverted repeat nationallicence TNP : Transposon insertion line nationallicence Singh Jaswinder Department of Plant and Microbial Biology, University of California, 94720, Berkeley, CA, USA aut Zhang Shibo Department of Plant and Microbial Biology, University of California, 94720, Berkeley, CA, USA aut Chen Calvin Department of Plant and Microbial Biology, University of California, 94720, Berkeley, CA, USA aut Cooper Laurel Department of Crop and Soil Sciences, Oregon State University, 97331, Corvallis, OR, USA aut Bregitzer Phil USDA-ARS, 83210, Aberdeen, ID, USA aut Sturbaum Anne USDA-ARS, 83210, Aberdeen, ID, USA aut Hayes Patrick Department of Crop and Soil Sciences, Oregon State University, 97331, Corvallis, OR, USA aut Lemaux Peggy Department of Plant and Microbial Biology, University of California, 94720, Berkeley, CA, USA aut Plant Molecular Biology Kluwer Academic Publishers 62/6(2006-12-01), 937-950 0167-4412 62:6<937 2006 62 11103 https://doi.org/10.1007/s11103-006-9067-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11103-006-9067-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Singh Jaswinder Department of Plant and Microbial Biology, University of California, 94720, Berkeley, CA, USA aut NATIONALLICENCE 700 1- Zhang Shibo Department of Plant and Microbial Biology, University of California, 94720, Berkeley, CA, USA aut NATIONALLICENCE 700 1- Chen Calvin Department of Plant and Microbial Biology, University of California, 94720, Berkeley, CA, USA aut NATIONALLICENCE 700 1- Cooper Laurel Department of Crop and Soil Sciences, Oregon State University, 97331, Corvallis, OR, USA aut NATIONALLICENCE 700 1- Bregitzer Phil USDA-ARS, 83210, Aberdeen, ID, USA aut NATIONALLICENCE 700 1- Sturbaum Anne USDA-ARS, 83210, Aberdeen, ID, USA aut NATIONALLICENCE 700 1- Hayes Patrick Department of Crop and Soil Sciences, Oregon State University, 97331, Corvallis, OR, USA aut NATIONALLICENCE 700 1- Lemaux Peggy Department of Plant and Microbial Biology, University of California, 94720, Berkeley, CA, USA aut NATIONALLICENCE 773 0- Plant Molecular Biology Kluwer Academic Publishers 62/6(2006-12-01), 937-950 0167-4412 62:6<937 2006 62 11103 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer