Plant & Animal Genomes XIII Conference Plant & Animal Genomes XIII ConferenceJanuary 15-19, 2005
Town & Country Convention Center
San Diego, CA
Jillian Perry1 , Tracey Welham2 , Trevor Wang2 , Martin Parniske3
The Sainsbury Laboratory and John Innes Centre have become the first training centre in Europe offering a new biotechnological approach to isolating plant mutants. This technology, pioneered by Steve Henikoff and his group (Mc Callum et al. 2000) in Seattle has been adapted in a collaboration between the two Norwich institutes for discovering mutants in known genes of the model plant, Lotus japonicus, a member of the legume family . The technology, known as TILLING (Targeting Induced Local Lesions IN Genomes), is a 'reverse genetics' process that relies on the ability of a special enzyme to detect mismatches in normal and mutant DNA strands when they are annealed.
Since June 2003 the Lotus japonicus TILLING facility (Perry et al. 2003) has been available to the research community. We have three populations available for immediate screening. A TILLING population of approximately 5000 M2 plants, a second population of 616 pre-screened symbiosis-defective plants and a third population of 96 putative starch mutants.
Our initial results have shown that the main TILLING population has on average 1302 mutations per genome (haploid Lotus genome ~500 Mb Hayashi et al. 2001). From the first 17 ca. 1kb fragments, 97 mutations were identified of which 1% were truncations, 64% resulted in missense changes and 35% caused silent changes. Additionally, 29 1-1.5kb fragments from 15 genes with known or suspected role in symbiosis have been screened using the symbiosis defective population (616 plants), identifying 64 mutations. Fourteen percent of these mutations were truncations, 62.5% missense mutations and 23.5% silent mutations.