Identification of Transposon Insertion Sites in Maize Mu-Tagged Mutants Using Mu-Seq

Area of Science:

• Plant genetics
• Genomics
• Molecular biology

Background:

• Mutator (Mu) transposons are mobile genetic elements used for insertional mutagenesis in maize.
• Resources like the BonnMu population facilitate functional genomics studies by providing Mu-tagged maize lines.
• Identifying transposon insertion sites is crucial for understanding gene function and genotype-specific mutations.

Purpose of the Study:

• To provide a detailed protocol for the Mutant-Seq (Mu-Seq) method.
• To enable high-throughput identification and mapping of Mu insertion sites in maize.
• To support the development of customized Mu-tagged mutant collections for genetic research.

Main Methods:

• Generation of multiplexed sequencing libraries (Mu-Seq libraries) from Mu-tagged maize populations.
• High-throughput sequencing of the constructed libraries.
• Data processing using the Mu-Seq Workflow Utility (MuWU) tool for precise mapping of Mu insertion sites.

Main Results:

• A comprehensive protocol for Mu-Seq, covering mutant population generation to data analysis.
• Demonstration of Mu-Seq as an effective high-throughput method for mapping Mu insertion sites.
• Establishment of a workflow for identifying genotype-specific mutations and candidate genes.

Conclusions:

• The Mu-Seq method provides a powerful tool for high-throughput mapping of Mu insertion sites in maize.
• This protocol facilitates the creation of tailored mutant collections for diverse genetic backgrounds.
• Mu-Seq aids in characterizing genotype-specific mutations and identifying genes associated with mutant phenotypes, advancing maize functional genomics.