Acknowledgments
We are grateful to R. Moscatiello and S. Sello, previous members of
our laboratory (Padova, Italy), for their contribution to the establishment of the described experimental procedures. We also thank
I. Szabo (Padova, Italy) for useful discussions and T. Morosinotto
(Padova, Italy) for the availability of the PAM imaging apparatus.
This work was supported by Progetti di Ricerca di Interesse Dipartimentale (PRID 2018) (prot. BIRD180317) and Dotazione Ordinaria della Ricerca 2018-2019 to LN.
References
1. Loewenberg JR (1965) Callus cultures of Arabidopsis. Arabidopsis Inf Serv 2:34
2. Negrutiu I, Jacobs M (1975) Arabidopsis thaliana as a model system in somatic cell genetics
II. Cell suspension culture. Plant Sci Lett
8:7–15
3. Moscatiello R, Baldan B, Navazio L (2013)
Plant cell suspension cultures. Methods Mol
Biol 953:77–93
4. Barkla BJ, Vera-Estrella R, Pantoja O (2014)
Growing Arabidopsis in vitro: cell suspensions,
in vitro culture, and regeneration. Methods
Mol Biol 1062:53–62
5. Hampp C, Richter A, Osorio S et al (2012)
Establishment of a photoautotrophic cell suspension culture of Arabidopsis thaliana for
photosynthetic, metabolic, and signaling studies. Mol Plant 5:524–527
6. Sello S, Moscatiello R, La Rocca N et al (2017)
A rapid and efficient method to obtain photosynthetic cell suspension cultures of Arabidopsis thaliana. Front Plant Sci 8:1444
7. Gutie ´rrez J, Gonza ´lez-Pe ´rez S, Garcı ´a-Garcı ´a F
et al (2014) Programmed cell death activated
by Rose Bengal in Arabidopsis thaliana cell
suspension cultures requires functional chloroplasts. J Exp Bot 65:3081–3095
8. Sello S, Perotto J, Carraretto L et al (2016)
Dissecting stimulus-specific Ca
2+ signals in
amyloplasts and chloroplasts of Arabidopsis
thaliana cell suspension cultures. J Exp Bot
67:3965–3974
9. Sello S, Moscatiello R, Mehlmer N et al (2018)
Chloroplast Ca
2+ fluxes into and across thylakoids revealed by thylakoid-targeted aequorin
probes. Plant Physiol 177:38–51
10. Choi WG, Miller G, Wallace I et al (2017)
Orchestrating rapid long-distance signaling in
plants with Ca
2+ , ROS and electrical signals.
Plant J 90:698–707
11. Hilleary R, Gilroy S (2018) Systemic signaling
in response to wounding and pathogens. Curr
Opin Plant Biol 43:57–62
12. Zeng Y, Ji C, Lin Y, Jiang L (2020) Transient
expression of fluorescent fusion proteins in
Arabidopsis protoplasts. In: Sa ´nchez-Serrano
JJ and Salinas J (eds) Arabidopsis Protocols.
Methods in Molecular Biology, vol 2200.
Springer
13. Teardo E, Carraretto L, Moscatiello R et al
(2019) A chloroplast-localized mitochondrial
calcium uniporter transduces osmotic stress in
Arabidopsis. Nat Plants 5:581–588
14. Jarvis P, Lo ´ pez-Juez E (2013) Biogenesis and
homeostasis of chloroplasts and other plastids.
Nat Rev Mol Cell Biol 14:787–802
15. Sadali NM, Sowden RG, Ling Q, Jarvis RP
(2019) Differentiation of chromoplasts and
other plastids in plants. Plant Cell Rep
38:803–818
16. Otegui MS (2020) Electron microscopy for
ultrastructure determination. In: Sa ´nchez-Serrano JJ and Salinas J (eds) Arabidopsis Protocols. Methods in Molecular Biology, vol 2200.
Springer
17. Calhoun C, Crist D, Knee E et al (2020)
Handling Arabidopsis and other Brassicaceae:
growth, preservation of seeds, transformation
and genetic crosses. In: Sa ´nchez-Serrano JJ and
Salinas J (eds) Arabidopsis Protocols. Methods
in Molecular Biology, vol 2200. Springer
Cell Suspension Cultures
185
We are grateful to R. Moscatiello and S. Sello, previous members of
our laboratory (Padova, Italy), for their contribution to the establishment of the described experimental procedures. We also thank
I. Szabo (Padova, Italy) for useful discussions and T. Morosinotto
(Padova, Italy) for the availability of the PAM imaging apparatus.
This work was supported by Progetti di Ricerca di Interesse Dipartimentale (PRID 2018) (prot. BIRD180317) and Dotazione Ordinaria della Ricerca 2018-2019 to LN.
References
1. Loewenberg JR (1965) Callus cultures of Arabidopsis. Arabidopsis Inf Serv 2:34
2. Negrutiu I, Jacobs M (1975) Arabidopsis thaliana as a model system in somatic cell genetics
II. Cell suspension culture. Plant Sci Lett
8:7–15
3. Moscatiello R, Baldan B, Navazio L (2013)
Plant cell suspension cultures. Methods Mol
Biol 953:77–93
4. Barkla BJ, Vera-Estrella R, Pantoja O (2014)
Growing Arabidopsis in vitro: cell suspensions,
in vitro culture, and regeneration. Methods
Mol Biol 1062:53–62
5. Hampp C, Richter A, Osorio S et al (2012)
Establishment of a photoautotrophic cell suspension culture of Arabidopsis thaliana for
photosynthetic, metabolic, and signaling studies. Mol Plant 5:524–527
6. Sello S, Moscatiello R, La Rocca N et al (2017)
A rapid and efficient method to obtain photosynthetic cell suspension cultures of Arabidopsis thaliana. Front Plant Sci 8:1444
7. Gutie ´rrez J, Gonza ´lez-Pe ´rez S, Garcı ´a-Garcı ´a F
et al (2014) Programmed cell death activated
by Rose Bengal in Arabidopsis thaliana cell
suspension cultures requires functional chloroplasts. J Exp Bot 65:3081–3095
8. Sello S, Perotto J, Carraretto L et al (2016)
Dissecting stimulus-specific Ca
2+ signals in
amyloplasts and chloroplasts of Arabidopsis
thaliana cell suspension cultures. J Exp Bot
67:3965–3974
9. Sello S, Moscatiello R, Mehlmer N et al (2018)
Chloroplast Ca
2+ fluxes into and across thylakoids revealed by thylakoid-targeted aequorin
probes. Plant Physiol 177:38–51
10. Choi WG, Miller G, Wallace I et al (2017)
Orchestrating rapid long-distance signaling in
plants with Ca
2+ , ROS and electrical signals.
Plant J 90:698–707
11. Hilleary R, Gilroy S (2018) Systemic signaling
in response to wounding and pathogens. Curr
Opin Plant Biol 43:57–62
12. Zeng Y, Ji C, Lin Y, Jiang L (2020) Transient
expression of fluorescent fusion proteins in
Arabidopsis protoplasts. In: Sa ´nchez-Serrano
JJ and Salinas J (eds) Arabidopsis Protocols.
Methods in Molecular Biology, vol 2200.
Springer
13. Teardo E, Carraretto L, Moscatiello R et al
(2019) A chloroplast-localized mitochondrial
calcium uniporter transduces osmotic stress in
Arabidopsis. Nat Plants 5:581–588
14. Jarvis P, Lo ´ pez-Juez E (2013) Biogenesis and
homeostasis of chloroplasts and other plastids.
Nat Rev Mol Cell Biol 14:787–802
15. Sadali NM, Sowden RG, Ling Q, Jarvis RP
(2019) Differentiation of chromoplasts and
other plastids in plants. Plant Cell Rep
38:803–818
16. Otegui MS (2020) Electron microscopy for
ultrastructure determination. In: Sa ´nchez-Serrano JJ and Salinas J (eds) Arabidopsis Protocols. Methods in Molecular Biology, vol 2200.
Springer
17. Calhoun C, Crist D, Knee E et al (2020)
Handling Arabidopsis and other Brassicaceae:
growth, preservation of seeds, transformation
and genetic crosses. In: Sa ´nchez-Serrano JJ and
Salinas J (eds) Arabidopsis Protocols. Methods
in Molecular Biology, vol 2200. Springer
Cell Suspension Cultures
185