Chapter 18
Identification and Characterization of Reproductive
Mutations in Arabidopsis
Marı ´a Flores-Tornero, Stefanie Sprunck, and Thomas Dresselhaus
Abstract
Mutations in numerous genes affect reproduction in Arabidopsis leading to sterility and abortion of seed
development, respectively. These include mutations in regulators of reproductive development and fertilization, but also in house-keeping genes lacking mutant phenotypes during vegetative development.
However, during the haploid phase of germline development or during seed development, lethality or
failures become visible when gene activity is needed. Plant reproduction is complex and includes many
processes from flowering and flower organ development toward the formation of seeds after a double
fertilization process. For those who are less familiar with the various reproductive processes in Arabidopsis
and who aim to study the cause of reproductive defects during germline development and function,
fertilization, or embryogenesis in a given mutant, we provide here a step-by-step guideline and basic
protocols to elucidate the reproductive process affected.
Key words Reproduction, Embryo sac, Pollen, Gamete, Fertilization, Embryogenesis, Arabidopsis
1 Introduction
Numerous mutations affect reproduction leading to defects in
flowering, flower organ and germline development, fertilization
mechanisms and seed development. In the model plant Arabidopsis
thaliana, failures in any of the above processes can be easily recognized by the formation of less and shorter siliques as well as by
reduced seed set. However, a detailed study is necessary to elucidate
the specific reproductive process being affected.
If shorter siliques are observed (Fig. 1a), we recommend to
initially study flower morphology. Arabidopsis flowers consist of
four sepals, four petals, two short and four long stamens, and two
fused female carpels forming the gynoecium that contains about
50–60 ovules (Fig. 1b). Defects in flower organ development (e.g.,
short stamens like in Fig. 1c) likely prevent self-pollination and thus
lead to reduced silique length indicating sterility (Fig. 1d). At
optimal growing conditions siliques reach a length of up to
Jose J. Sanchez-Serrano and Julio Salinas (eds.), Arabidopsis Protocols, Methods in Molecular Biology, vol. 2200,
https://doi.org/10.1007/978-1-0716-0880-7_18, © Springer Science+Business Media, LLC, part of Springer Nature 2021
371
Identification and Characterization of Reproductive
Mutations in Arabidopsis
Marı ´a Flores-Tornero, Stefanie Sprunck, and Thomas Dresselhaus
Abstract
Mutations in numerous genes affect reproduction in Arabidopsis leading to sterility and abortion of seed
development, respectively. These include mutations in regulators of reproductive development and fertilization, but also in house-keeping genes lacking mutant phenotypes during vegetative development.
However, during the haploid phase of germline development or during seed development, lethality or
failures become visible when gene activity is needed. Plant reproduction is complex and includes many
processes from flowering and flower organ development toward the formation of seeds after a double
fertilization process. For those who are less familiar with the various reproductive processes in Arabidopsis
and who aim to study the cause of reproductive defects during germline development and function,
fertilization, or embryogenesis in a given mutant, we provide here a step-by-step guideline and basic
protocols to elucidate the reproductive process affected.
Key words Reproduction, Embryo sac, Pollen, Gamete, Fertilization, Embryogenesis, Arabidopsis
1 Introduction
Numerous mutations affect reproduction leading to defects in
flowering, flower organ and germline development, fertilization
mechanisms and seed development. In the model plant Arabidopsis
thaliana, failures in any of the above processes can be easily recognized by the formation of less and shorter siliques as well as by
reduced seed set. However, a detailed study is necessary to elucidate
the specific reproductive process being affected.
If shorter siliques are observed (Fig. 1a), we recommend to
initially study flower morphology. Arabidopsis flowers consist of
four sepals, four petals, two short and four long stamens, and two
fused female carpels forming the gynoecium that contains about
50–60 ovules (Fig. 1b). Defects in flower organ development (e.g.,
short stamens like in Fig. 1c) likely prevent self-pollination and thus
lead to reduced silique length indicating sterility (Fig. 1d). At
optimal growing conditions siliques reach a length of up to
Jose J. Sanchez-Serrano and Julio Salinas (eds.), Arabidopsis Protocols, Methods in Molecular Biology, vol. 2200,
https://doi.org/10.1007/978-1-0716-0880-7_18, © Springer Science+Business Media, LLC, part of Springer Nature 2021
371