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 F1 plants 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 10-4 and 10-5 per F1 plant. 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 F1 plants, 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.
May
Bruce P
BP
Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA.
Liu
Hong
H
Vollbrecht
Erik
E
Senior
Lynn
L
Rabinowicz
Pablo D
PD
Roh
Donna
D
Pan
Xiaokang
X
Stein
Lincoln
L
Freeling
Mike
M
Alexander
Danny
D
Martienssen
Rob
R
D001483:Base Sequence; D017931:DNA Primers; D004251:DNA Transposable Elements; D016254:Mutagenesis, Insertional; D010641:Phenotype; D016133:Polymerase Chain Reaction; D003313:Zea mays / Q000235:genetics*
D017931:DNA Primers; D004251:DNA Transposable Elements