Abstract
Seeds of rapid-cycling Brassica oleracea were mutagenized with the chemical mutagen, ethylmethane sulfonate. The reverse genetics technique, TILLING, was used on a sample population of 1,000 plants, to determine the mutation profile. The spectrum and frequency of mutations induced by ethylmethane sulfonate was similar to that seen in other diploid species such as Arabidopsis thaliana. These data indicate that the mutagenesis was effective and demonstrate that TILLING represents an efficient reverse genetic technique in B. oleracea that will become more valuable as increasing genomic sequence data become available for this species. The extensive duplication in the B. oleracea genome is believed to result in the genetic redundancy that has been important for the evolution of morphological diversity seen in today’s B. oleracea crops (broccoli, Brussels sprouts, cauliflower, cabbage, kale and kohlrabi). However, our forward genetic screens identified 120 mutants in which some aspect of development was affected. Some of these lines have been characterized genetically and in the majority of these, the mutant trait segregates as a recessive allele affecting a single locus. One dominant mutation (curly leaves) and one semi-dominant mutation (dwarf-like) were also identified. Allelism tests of two groups of mutants (glossy and dwarf) revealed that for some loci, multiple independent alleles have been identified. These data indicate that, despite genetic redundancy, mutation of many individual loci in B. oleracea results in distinct phenotypes.
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Acknowledgments
We are grateful to Quentin Cronk and the Faculty of Land and Food Systems at UBC for providing laboratory space for CAN-TILL. Funding for the TILLING part of this project was from a Genome Prairie/Genome Canada Functional Genomics of Abiotic Stress grant to GWH and IAPP. Funding for the generation of mutants and forward genetic screening was from US Department of Agriculture Initiative for Future Agriculture and Food Systems. Himelblau and Mentzer are supported by the NSF Plant Genome Program (DBI-0077774).
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Communicated by C. F. Quiros.
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Himelblau, E., Gilchrist, E.J., Buono, K. et al. Forward and reverse genetics of rapid-cycling Brassica oleracea . Theor Appl Genet 118, 953–961 (2009). https://doi.org/10.1007/s00122-008-0952-7
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DOI: https://doi.org/10.1007/s00122-008-0952-7