TY - CHAP
T1 - Workflow to characterize mutants with reproductive defects
AU - Noble, Jennifer A.
AU - Palanivelu, Ravishankar
N1 - Funding Information:
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).
Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2020.
PY - 2020
Y1 - 2020
N2 - 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.
AB - 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.
KW - Aniline blue staining
KW - Fertility
KW - Male gametophyte and sporophyte
KW - Ovules
KW - Pistil
KW - Pollen
KW - Pollen tube
KW - Pollen tube–pistil interactions
KW - Seed set
KW - Transmission efficiency
KW - Transmission efficiency
UR - http://www.scopus.com/inward/record.url?scp=85086355189&partnerID=8YFLogxK
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U2 - 10.1007/978-1-0716-0672-8_8
DO - 10.1007/978-1-0716-0672-8_8
M3 - Chapter
C2 - 32529432
AN - SCOPUS:85086355189
T3 - Methods in Molecular Biology
SP - 109
EP - 128
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -