A mutant Brassica napus (canola) population for the identification of new genetic diversity via TILLING and next generation sequencing.

ABSTRACT: We have generated a Brassica napus (canola) population of 3,158 EMS-mutagenised lines and used TILLING to demonstrate that the population has a high enough mutation density that it will be useful for identification of mutations in genes of interest in this important crop species. TILLING is a reverse genetics technique that has been successfully used in many plant and animal species. Classical TILLING involves the generation of a mutagenised population, followed by screening of DNA samples using a mismatch-specific endonuclease that cleaves only those PCR products that carry a mutation. Polyacrylamide gel detection is then used to visualise the mutations in any gene of interest. We have used this TILLING technique to identify 432 unique mutations in 26 different genes in B. napus (canola c

SUBMITTER: Gilchrist EJ 

PROVIDER: S-EPMC3869819 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature