Plant, Animal & Microbe Genomes X Conference Plant, Animal & Microbe Genomes X ConferenceJanuary 12-16, 2002
Town & Country Convention Center
San Diego, CA
Workshop: Barley
Insertional mutagenesis using transposable elements has become a valuable tool for functional genomics in many plant species. So far, the systematic use of insertional mutagenesis in monocots has been limited to maize and rice. Barley serves as a model of the Triticeae family because it is the only true diploid self-compatible species for which a large number of genetic stocks and genome tools are available. In order to develop a two-element system for insertional mutagenesis in barley, we introduced a stabilised Ac-Transposase gene and a Ds-element from maize into barley. Crossing with Ac-expressing plants activated transpositions of the non-autonomous Ds-element and the transpositional behaviour of Ds was extensively characterised. Populations of Ds-containing plants were developed that can be used effectively for different gene-tagging approaches. Large numbers of independent single copy Ds-containing lines have been generated and can be used for targeted gene-tagging strategies after mapping the position of Ds. In order to obtain a sufficient coverage of the barley genome with launch pads for targeted tagging, the production of further independent single-copy transgenic barley lines is in progress. In a second approach a population of plants carrying multiple independent Ds inserts has been set up. Sequence analysis of isolated Ds flanking regions showed that 40 % of the insertion sites have high similarity to EST's from barley or other cereals. A tagged insertion site and a mapped insertion site database will be established and made public for researchers using reverse genetics approaches to determine gene function in barley.