Abstract
Reverse genetics approaches for characterizing phenotypes of mutants in a gene of interest (GOI) require thorough genotyping and phenotypic analysis. However, special challenges are encountered when a GOI is expressed in reproductive tissues: a variety of assays are required to characterize the phenotype and a mutant may show sporophytic and/or gametophytic defects in male and/or female reproductive tissues, which are structurally and functionally intertwined. Here, we present a streamlined workflow to characterize mutants with reproductive defects, primarily using Arabidopsis as a model, which can also be adapted to characterize mutants in other flowering plants. Procedures described here can be used to distinguish different kinds of reproductive defects and pinpoint the defective reproductive step(s) in a mutant. Although our procedures emphasize the characterization of mutants with male reproductive defects, they can nevertheless be used to identify female reproductive defects, as those defects could manifest alongside, and sometimes require, male reproductive tissues.
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Acknowledgements
J.A. Noble was supported by the following: IGERT Comparative Genomics Program at the University of Arizona (Award ID: 0654435); NSF Graduate Research Fellowship: Grant DGE-1143953; the Boynton Graduate Fellowship from the School of Plant Sciences, University of Arizona; and the University of Arizona Graduate College Office of Diversity and Inclusion. Additional support for this work was provided by an NSF grant to R. Palanivelu (IOS-1146090).
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Noble, J.A., Palanivelu, R. (2020). Workflow to Characterize Mutants with Reproductive Defects. In: Geitmann, A. (eds) Pollen and Pollen Tube Biology. Methods in Molecular Biology, vol 2160. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0672-8_8
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DOI: https://doi.org/10.1007/978-1-0716-0672-8_8
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