Leaf phenomics: a systematic reverse genetic screen for Arabidopsis leaf mutants

Area of Science:

• Plant Biology
• Genetics
• Molecular Biology

Background:

• Understanding leaf organogenesis is crucial for plant development and manipulation.
• Previous forward genetic screens in Arabidopsis identified many leaf development mutants, but the genome remains unscreened.

Purpose of the Study:

• To identify novel genes involved in leaf development by screening the Arabidopsis Salk Unimutant collection.
• To develop a comprehensive resource for studying leaf development and facilitating gene discovery.

Main Methods:

• Screening of the Arabidopsis Salk Unimutant collection for leaf development phenotypes.
• Phenotypic characterization using a custom leaf phenotype ontology.
• Genotyping and T-DNA insertion mapping using whole-genome sequencing.
• Development of the public web application PhenoLeaf for data access.

Main Results:

• Identified 608 lines with full penetrance leaf phenotypes and 98 with segregating phenotypes.
• Confirmed T-DNA insertion in annotated loci for 78% of genotyped mutants.
• Developed a cost-effective whole-genome sequencing method for mapping non-annotated insertions.
• Characterized two novel genes, At1g77600 and At3g62870, involved in leaf growth and function.

Conclusions:

• The screened collection and PhenoLeaf web application provide a valuable resource for plant genetics research.
• Facilitated discovery of genes essential for proximal-distal leaf growth, cell proliferation, and chloroplast function.
• Supports research in biomass feedstocks and other traits of this fundamental photosynthetic organ.