Maize-Targeted Mutagenesis: A Knockout Resource for Maize

We describe an efficient system for site-selected transposon mutagenesis in maize. A total of 43,776 F1plants were generated by using Robertson's Mutator (Mu) pollen parents and self-pollinated to establish a library of transposon-mutagenized seed. The frequency of new seed mutants was between...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 100; no. 20; pp. 11541 - 11546
Main Authors May, Bruce P., Liu, Hong, Vollbrecht, Erik, Senior, Lynn, Rabinowicz, Pablo D., Roh, Donna, Pan, Xiaokang, Stein, Lincoln, Freeling, Mike, Alexander, Danny, Martienssen, Rob
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 30.09.2003
National Acad Sciences
Subjects
Base Sequence
Biological Sciences
Corn
Data analysis
DNA
DNA Primers
DNA Transposable Elements
Genetic mutation
Genetic screening
Genetics
Genomics
Mutagenesis, Insertional
Mutation
Phenotype
Phenotypes
Polymerase Chain Reaction
Reverse genetics
Tissues
Transposons
Zea mays - genetics
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Summary:We describe an efficient system for site-selected transposon mutagenesis in maize. A total of 43,776 F1plants were generated by using Robertson's Mutator (Mu) pollen parents and self-pollinated to establish a library of transposon-mutagenized seed. The frequency of new seed mutants was between 10-4and 10-5per F1plant. As a service to the maize community, maize-targeted mutagenesis selects insertions in genes of interest from this library by using the PCR. Pedigree, knockout, sequence, phenotype, and other information is stored in a powerful interactive database (maize-targeted mutagenesis database) that enables analysis of the entire population and the handling of knockout requests. By inhibiting Mu activity in most F1plants, we sought to reduce somatic insertions that may cause false positives selected from pooled tissue. By monitoring the remaining Mu activity in the F2, however, we demonstrate that seed phenotypes depend on it, and false positives occur in lines that appear to lack it. We conclude that more than half of all mutations arising in this population are suppressed on losing Mu activity. These results have implications for epigenetic models of inbreeding and for functional genomics.
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This paper was submitted directly (Track II) to the PNAS office.
Abbreviations: MTM, maize-targeted mutagenesis; Mu, Mutator; mtmDB, MTM database.
To whom correspondence should be addressed. E-mail: martiens@cshl.org.
B.P.M., H.L., and E.V. contributed equally to this work.
Edited by Ronald L. Phillips, University of Minnesota, St. Paul, MN, and approved July 14, 2003
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1831119100