title: A PCR-based forward genetics screening, using expression domain-specific markers, identifies mutants in endosperm transfer cell development.
abstract: Mutant collections are an invaluable source of material on which forward genetic approaches allow the identification of genes affecting a wide variety of biological processes. However, some particular developmental stages and morphological structures may resist analysis due to their physical inaccessibility or to deleterious effects associated to their modification. Furthermore, lethal mutations acting early in development may escape detection. We have approached the characterization of 101 maize seed mutants, selected from a collection of 27,500 visually screened Mu-insertion lines, using a molecular marker approach based on a set of genes previously ascribed to different tissue compartments within the early developing kernel. A streamlined combination of qRT-PCR assays has allowed us to preliminary pinpoint the affected compartment, establish developmental comparisons to WT siblings and select mutant lines with alterations in the different compartments. Furthermore, clusters of markers co-affected by the underlying mutation were identified. We have analyzed more extensively a set of lines presenting significant variation in transfer cell-associated expression markers, and have performed morphological observations, and immunolocalization experiments to confirm the results, validating this approach as an efficient mutant description tool.
authors.affiliation: Departamento Biomedicina and Biotecnología (Genética), Universidad de Alcalá Alcalá de Henares, Spain.
authors.forename: Luis M
authors.initials: LM
authors.lastname: Muñiz
authors.affiliation: Departamento Biomedicina and Biotecnología (Genética), Universidad de Alcalá Alcalá de Henares, Spain.
authors.forename: Elisa
authors.initials: E
authors.lastname: Gómez
authors.affiliation: GM Trait Discovery, Biogemma, Centre de Recherche de Chappes Chappes, France.
authors.forename: Virginie
authors.initials: V
authors.lastname: Guyon
authors.affiliation: Departamento Biomedicina and Biotecnología (Genética), Universidad de Alcalá Alcalá de Henares, Spain.
authors.forename: Maribel
authors.initials: M
authors.lastname: López
authors.affiliation: GM Trait Discovery, Biogemma, Centre de Recherche de Chappes Chappes, France.
authors.forename: Bouchaib
authors.initials: B
authors.lastname: Khbaya
authors.affiliation: GM Trait Discovery, Biogemma, Centre de Recherche de Chappes Chappes, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Sellam
authors.affiliation: GM Trait Discovery, Biogemma, Centre de Recherche de Chappes Chappes, France.
authors.forename: Pascual
authors.initials: P
authors.lastname: Peréz
authors.affiliation: Departamento Biomedicina and Biotecnología (Genética), Universidad de Alcalá Alcalá de Henares, Spain.
authors.forename: Gregorio
authors.initials: G
authors.lastname: Hueros
keywords: aleurone; development; gene expression; mutant; transfer cells