Chapter 1
Handling Arabidopsis and Other Brassicaceae: Growth,
Preservation of Seeds, Transformation, and Genetic Crosses
Christopher Calhoun, Deborah Crist, Emma Knee, Julie Miller, Eva Nagy,
and David E. Somers
Abstract
Achieving optimal plant growth is essential for the advancement of Arabidopsis thaliana (Arabidopsis)
research. Over the last 20 years, the Arabidopsis Biological Resource Center (ABRC) has collected and
developed a series of best-practice protocols, some of which are presented in this chapter. Arabidopsis can
be grown in a variety of locations, growth media, and environmental conditions. Some mutant genotypes,
natural accessions, and Arabidopsis relatives require strictly controlled growth conditions best provided by
growth rooms, chambers, or incubators. Other lines can be grown in less-controlled greenhouse settings.
Although the majority of lines can be grown in soil, certain experimental purposes require utilization of
sterile solid or liquid growth media. These include the selection of primary transformants, identification of
homozygous lethal individuals in a segregating population, or bulking of a large amount of plant material.
The importance of controlling, observing, and recording growth conditions is emphasized and appropriate
equipment for monitoring these conditions is listed. Proper conditions for seed harvest and preservation, as
well as seed quality control procedures, are also described. In addition, plant transformation and genetic
crosses, two of the methods that revolutionized Arabidopsis genetics, are discussed.
Key words Arabidopsis, Growth conditions, Environmental conditions, Natural accession, Seed
germination, Seed quality, Plant transformation, Genetic crosses
1 Introduction
Healthy growth and development of plants is a prerequisite for
accurate and reproducible plant research and Arabidopsis thaliana
(Arabidopsis) is no exception. Proper handling and maintenance of
Arabidopsis plants is also required for optimal seed production. In
this chapter, we describe basic, best-practice protocols for handling
Arabidopsis. The reader should be aware that common growth
conditions, particularly in greenhouses, may not be similar to
those of the native habitats of natural accessions. This is especially
important when interpreting phenotypic differences in traits that
are known to be strongly influenced by environmental conditions,
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_1, © Springer Science+Business Media, LLC, part of Springer Nature 2021
3
Handling Arabidopsis and Other Brassicaceae: Growth,
Preservation of Seeds, Transformation, and Genetic Crosses
Christopher Calhoun, Deborah Crist, Emma Knee, Julie Miller, Eva Nagy,
and David E. Somers
Abstract
Achieving optimal plant growth is essential for the advancement of Arabidopsis thaliana (Arabidopsis)
research. Over the last 20 years, the Arabidopsis Biological Resource Center (ABRC) has collected and
developed a series of best-practice protocols, some of which are presented in this chapter. Arabidopsis can
be grown in a variety of locations, growth media, and environmental conditions. Some mutant genotypes,
natural accessions, and Arabidopsis relatives require strictly controlled growth conditions best provided by
growth rooms, chambers, or incubators. Other lines can be grown in less-controlled greenhouse settings.
Although the majority of lines can be grown in soil, certain experimental purposes require utilization of
sterile solid or liquid growth media. These include the selection of primary transformants, identification of
homozygous lethal individuals in a segregating population, or bulking of a large amount of plant material.
The importance of controlling, observing, and recording growth conditions is emphasized and appropriate
equipment for monitoring these conditions is listed. Proper conditions for seed harvest and preservation, as
well as seed quality control procedures, are also described. In addition, plant transformation and genetic
crosses, two of the methods that revolutionized Arabidopsis genetics, are discussed.
Key words Arabidopsis, Growth conditions, Environmental conditions, Natural accession, Seed
germination, Seed quality, Plant transformation, Genetic crosses
1 Introduction
Healthy growth and development of plants is a prerequisite for
accurate and reproducible plant research and Arabidopsis thaliana
(Arabidopsis) is no exception. Proper handling and maintenance of
Arabidopsis plants is also required for optimal seed production. In
this chapter, we describe basic, best-practice protocols for handling
Arabidopsis. The reader should be aware that common growth
conditions, particularly in greenhouses, may not be similar to
those of the native habitats of natural accessions. This is especially
important when interpreting phenotypic differences in traits that
are known to be strongly influenced by environmental conditions,
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_1, © Springer Science+Business Media, LLC, part of Springer Nature 2021
3