Abstract
Arabidopsis thaliana research relies heavily on the use of in vitro culture media for phenotypic characterization of seedling responses to intrinsic and extrinsic signals. For this, the most favorable growth conditions must be established and used as a reference, especially in those studies aimed at characterizing responses to abiotic and biotic stresses. Standard in vitro protocols commonly used for the growth and characterization of A. thaliana plants show suboptimal composition due to excessive nutrient content, representing an abiotic stress per se. We study here the nutritional factors that affect in vitro growth of A. thaliana seedlings and describe an optimized and nutritionally balanced culture medium. We show that this medium is appropriate for the growth and propagation of many A. thaliana mutants, including those that are unable to complete the life cycle because they lack the root system. The described method avoids bias in phenotypic characterization during abiotic/biotic stress experiments. This protocol makes it possible to complete the life cycle in 40–45 days and obtain sufficient seeds without the need for seed sterilization, avoiding the use of soil and saving space and time.
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Data Availability
The data supporting the findings of this study are available from the corresponding author, T. Pasternak, upon request.
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Acknowledgements
The authors thank Katja Rapp for many stimulating ideas and excellent technical assistance.
Funding
This work was supported by Bundesministerium für Bildung und Forschung (BMBF Microsystems), the Excellence Initiative of the German Federal and State Governments (EXC 294), and SFB746 an Deutsches Zentrum für Luft und Raumfahrt (DLR 50WB1022). TP holds a “María Zambrano” distinguished research contract at the UMH.
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TP performed the experiments; TP, JMP-P, TA, BR, and KP interpreted and discussed the results; TP, JMP-P, and TA wrote the manuscript.
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Pasternak, T., Pérez-Pérez, J.M., Ruperti, B. et al. A New In Vitro Growth System for Phenotypic Characterization and Seed Propagation of Arabidopsis thaliana. J Plant Growth Regul 43, 652–658 (2024). https://doi.org/10.1007/s00344-023-11093-x
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DOI: https://doi.org/10.1007/s00344-023-11093-x