NSF Award
0211732
There exist numerous genome-scale methods for obtaining mutations useful for functional analysis. In traditional genetics, random mutagenesis is followed by phenotypic analysis. However, the availability of large-scale genomic and cDNA sequence data encourages reverse genetic approaches. In plants, reverse genetics based on transposon mutagenesis or directed anti-sense inhibition have succeeded in some cases, but the methods can be labor-intensive and unreliable.<br/><br/>TILLING is a new reverse genetic strategy that combines random chemical mutagenesis with PCR-based screening of gene regions of interest. This provides a range of allele types, including missense and knock-out mutations, which are potentially useful in a variety of gene function and interaction studies. TILLING is especially suitable for plants, even for those that lack well-developed genetic tools. A low-cost high-throughput TILLING method has been developed and applied to screening mutagenized Arabidopsis populations. This effort has resulted in the Arabidopsis TILLING Project, which provides allelic series of point mutations in genes of interest to members of the general Arabidopsis community. Concomitantly, the TILLING team also introduced interactive web-based tools needed to identify gene regions most suitable for TILLING, to design optimal PCR primers for mutational screening, and to analyze the resulting mutations. <br/><br/>The successful delivery of several hundred sequenced mutations in Arabidopsis genes encourages the application of TILLING technology to crop plants. One goal of this proposal is to establish a high-throughput rice TILLING service utilizing mutagenized rice populations from the International Rice Research Institute and others. A second goal is to utilize the TILLING facility and resources for dissemination of TILLING technology to the broader plant biology community.<br/>Accomplishment of these goals should contribute to the understanding of gene functions for both crop plants and model organisms, and to the improvement of the plant genomics infrastructure for research and education. TILLING technology has potential benefits for agriculture, where methods are needed for crop improvement that avoid the expensive regulatory process required for transgenics and that are widely acceptable to consumers.
