Generating novel allelic variation through Activator insertional mutagenesis in maize

Area of Science:

• Plant genetics
• Molecular biology
• Genomics

Background:

• The Activator (Ac) transposable element is a valuable tool for insertional mutagenesis in plants.
• Developing efficient mutagenesis platforms is crucial for gene discovery and characterization in maize.

Purpose of the Study:

• To establish an Ac-based mutagenesis platform for maize.
• To generate and characterize novel alleles of the pink scutellum1 (ps1) gene.

Main Methods:

• Large-scale mutagenesis targeting the maize ps1 locus using a donor Ac element.
• Genetic screens to identify heritable ps1 excision alleles.
• Molecular characterization of Ac transposition and excision events.

Main Results:

• 1092 Ac transposition events were recovered, yielding 17 novel ps1 alleles.
• 83 germinally heritable ps1 excision alleles were identified.
• Analysis revealed a position-dependent bias in excision and predominant 7- and 8-bp footprints.

Conclusions:

• The developed Ac-based platform effectively generates diverse ps1 allelic variation.
• The methods are adaptable for creating novel alleles at any Ac-tagged gene in maize.
• Footprint analysis provides insights into Ac excision mechanisms in maize.