Workflow to characterize mutants with reproductive defects

Jennifer A. Noble, Ravishankar Palanivelu

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages109-128
Number of pages20
DOIs
StatePublished - 2020

Publication series

NameMethods in Molecular Biology
Volume2160
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Aniline blue staining
  • Fertility
  • Male gametophyte and sporophyte
  • Ovules
  • Pistil
  • Pollen
  • Pollen tube
  • Pollen tube–pistil interactions
  • Seed set
  • Transmission efficiency
  • Transmission efficiency

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Noble, J. A., & Palanivelu, R. (2020). Workflow to characterize mutants with reproductive defects. In Methods in Molecular Biology (pp. 109-128). (Methods in Molecular Biology; Vol. 2160). Humana Press Inc.. https://doi.org/10.1007/978-1-0716-0672-8_8