Biochemistry

Abstract

CS-16-4 - Mutagenomics: a high throughput strategy for screening of mutant lines

Monday, July 16
4:18 PM - 4:38 PM

In a mutagenetic screen, often a cumbersome task is the identification of causal genes. Generally, this necessitates the creation of a mapping population where the co-segregation of the mutant genotype with genetic markers is assessed. The mapping process has been greatly accelerated with the use of mapping-by-sequencing methods. However, these methods require the user to generate a segregating population, requiring two generations of growth and the selection of plants with the mutant phenotype from the F2 population. Further, such approaches focus on small set of target genes at a time. We present a strategy, dubbed “mutagenomics,” for screening through uncharacterized, uncrossed mutant lines in a parallel fashion. This strategy has the potential to reduce the number of candidate genes from mutant lines without requiring the researcher to perform back-crosses. Mutagenomics is a two-stage, two-prong strategy. After resequencing the genome of mutant lines, the first stage is the removal of lines with mutations in known genes. The second stage is a dual strategy which assesses whether any genes are mutated more often than expected by chance (i.e. identification of multiple alleles) and whether any genes are found to be mutated in multiple lines of common descent. This method does not preclude the use of mapping-by-sequencing technologies or traditional marker-based mapping. Either method can be used to map mutations in lines where mutagenomics did not identify a candidate mutation.


          We have applied mutagenomics to mutant lines from a screen for hyposensitivity to the hormone cytokinin, a critical regulator of diverse set of biological processes in plants. A pool of 28 mutant lines have been sequenced and mutations in ahk4 and hy5 were identified as very strong candidates for causal mutations. We anticipate this strategy will accelerate the pace and utility of genetic screens.


 

Co-Authors

Joseph Kieber – UNC Chapel Hill

Charles Hodgens

UNC Chapel Hill

Presentation(s):

Send Email for Charles Hodgens


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CS-16-4 - Mutagenomics: a high throughput strategy for screening of mutant lines



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