Plant, Animal & Microbe Genomes X Conference Plant, Animal & Microbe Genomes X ConferenceJanuary 12-16, 2002
Town & Country Convention Center
San Diego, CA
Workshop: Barley
Progress in DNA sequencing has propelled a shift in genome analysis from structural to functional genomics (ie. the genome-driven systematic study of gene function). Rapid methods for ascertaining the function of large numbers of genes are therefore highly desirable. With the development of sensitive methods such as chemical or enzymatic cleavage of mismatched DNAs, critical extension PCR and dHPLC-HA for the detection of 'aberrant' DNA fragments in pooled samples, the use of chemical or physical mutagens to facilitate targeted gene inactivation has significant potential. Strategies deploying mutagens which induce random point or small deletion mutations coupled with a PCR-based DNA mismatch screen theoretically allow the detection of mutations in any specified target region. Inherent attractions include the ability to manipulate the mutagen and its dose (influencing mutation type and frequency) and to scale up or down easily. We are investigating the potential of chemical / physical mutagenesis to facilitate reverse genetics in barley. Four M1 test populations for three mutagens (Gamma, DEB, EMS) have been grown and seed from M1 plants from 4 different dose rates of each of the 3 mutagens (600 families) carried forward to the M2 generation (c. 36,000 plants). Mutation frequency (in M2 plants) has been calculated based on lethality, visible phenotypes and genome-wide mutagen-induced DNA sequence polymorphism. In parallel, mutation detection protocols are being evaluated at several pooling depths. The potential of the system for high throughput gene function analysis in barley will be presented.