Is it possible to find a method that shows a correspondence between a known mutation and the phenotype in barley wax less mutants?

Sammanfattning: The wax on the stem and leafs of the barley (Hordeum vulgare L.) plant has a protective function and is produced by the cells on the plant's outer surfaces. There are several available barley mutants where the wax production has been stopped. Four mutants are the single-site mutants cer-c.36, cer-q.42, ceru. 21 and the deletion mutant cer-cqu.724. We phenotyped 636 barley plants and collected leaf samples from all of them for DNA extraction. We used PCR with pre-designed primers to confirm that DNA had been extracted from all plants. Since the mutations in cer-c.36, cer-q.42 and cer-u.21 are already known we tried to identify the mutant plants by using restriction enzymes to cleave a PCR product on the mutant site. Then we designed specific nucleotide primers for PCR that only amplifies a region of either mutated plants or the wild type plants. This method gave some interesting results for the wild type cer-u.21 mutant. Finally, we looked at the triple mutant cer-cqu.724 and since it had not been sequenced yet we used primers for cer-c.36 and cer-u.21 to try to amplify the wild type DNA and then confirm the results by amplifying a separate sequence in the barley genome to make sure that there is DNA extracted from all plants. In the end we only obtained a result for the cer-u.21 wild typeHowever, it should be possible to get results for the other mutants if the PRC program is optimized.