Chapter 8
Arabidopsis Photosynthetic and Heterotrophic Cell
Suspension Cultures
Enrico Cortese, Luca Carraretto, Barbara Baldan, and Lorella Navazio
Abstract
Cell suspension cultures represent a widely used experimental tool suitable to perform a variety of structural
and physiological studies in a more simplified system compared to the organism in toto. In this chapter we
describe the methods routinely used in our laboratory to establish and maintain Arabidopsis photosynthetic
and heterotrophic cell suspension cultures, containing either chloroplasts or amyloplasts, respectively. The
use of these in vitro systems may allow to obtain insights into the unique features of chloroplasts versus
non-green plastids, as well as their integration in the structural and metabolic compartmentalization of the
plant cell.
Key words Arabidopsis, Seeds, Culture media, Phytohormones, In vitro growth, Calli, Cell suspension cultures, Chloroplasts, Amyloplasts, Protoplasts
1 Introduction
Arabidopsis in vitro cultures have been established a long time ago,
with the first reports dating back to the 60s and 70s of the last
century [1, 2]. Methodologies for the production of calli—clusters
of dedifferentiated cells growing on solid culture media—as well as
the setup of Arabidopsis cell suspension cultures—cells growing in
liquid culture media under constant shaking—have been described
in detail [e.g., [3, 4]]. In vitro technologies, allowing for the
dissection of the entire Arabidopsis plant into its elementary
units, represent an invaluable tool in basic research, and have
greatly contributed to advancing our knowledge of the cellular
and molecular biology, biochemistry, and physiology of this
model plant.
For a long time most techniques have allowed only for the
obtainment of Arabidopsis heterotrophic cell suspension cultures—i.e., containing amyloplasts—, whereas the production of
photosynthetic cell suspension cultures—i.e., containing chloroplasts—has proven more difficult to obtain. More recently,
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_8, © Springer Science+Business Media, LLC, part of Springer Nature 2021
167
Arabidopsis Photosynthetic and Heterotrophic Cell
Suspension Cultures
Enrico Cortese, Luca Carraretto, Barbara Baldan, and Lorella Navazio
Abstract
Cell suspension cultures represent a widely used experimental tool suitable to perform a variety of structural
and physiological studies in a more simplified system compared to the organism in toto. In this chapter we
describe the methods routinely used in our laboratory to establish and maintain Arabidopsis photosynthetic
and heterotrophic cell suspension cultures, containing either chloroplasts or amyloplasts, respectively. The
use of these in vitro systems may allow to obtain insights into the unique features of chloroplasts versus
non-green plastids, as well as their integration in the structural and metabolic compartmentalization of the
plant cell.
Key words Arabidopsis, Seeds, Culture media, Phytohormones, In vitro growth, Calli, Cell suspension cultures, Chloroplasts, Amyloplasts, Protoplasts
1 Introduction
Arabidopsis in vitro cultures have been established a long time ago,
with the first reports dating back to the 60s and 70s of the last
century [1, 2]. Methodologies for the production of calli—clusters
of dedifferentiated cells growing on solid culture media—as well as
the setup of Arabidopsis cell suspension cultures—cells growing in
liquid culture media under constant shaking—have been described
in detail [e.g., [3, 4]]. In vitro technologies, allowing for the
dissection of the entire Arabidopsis plant into its elementary
units, represent an invaluable tool in basic research, and have
greatly contributed to advancing our knowledge of the cellular
and molecular biology, biochemistry, and physiology of this
model plant.
For a long time most techniques have allowed only for the
obtainment of Arabidopsis heterotrophic cell suspension cultures—i.e., containing amyloplasts—, whereas the production of
photosynthetic cell suspension cultures—i.e., containing chloroplasts—has proven more difficult to obtain. More recently,
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_8, © Springer Science+Business Media, LLC, part of Springer Nature 2021
167