Abstract
Laser capture microdissection (LCM) has become a powerful technique that allows analyzing gene expression in specific target cells from complex tissues. Widely used in animal research, still few studies on plants have been carried out. We have applied this technique to the plant–nematode interaction by isolating feeding cells (giant cells; GCs) immersed inside complex swelled root structures (galls) induced by root-knot nematodes. For this purpose, a protocol that combines good morphology preservation with RNA integrity maintenance was developed, and successfully applied to Arabidopsis and tomato galls. Specifically, early developing GCs at 3 and 7 days post-infection (dpi) were analyzed; RNA from LCM GCs was amplified and used successfully for microarray assays.
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Acknowledgements
The authors thank K. Lindsey and J. Topping (University of Durham, UK) for the use of their equipment and for their advice during the development of the microdissection protocol. Work in the laboratory was supported by grants from the Ministry of Education (AGL2016-75287-R) and by the Castilla-La Mancha Government (SBPLY/17/180501/000287) to CE. M.B. was a recipient of a technologist contract from the University of Castilla-La Mancha and the Fondo Europeo de Desarrollo Regional (FEDER).
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Barcala, M., Fenoll, C., Escobar, C. (2021). Laser Microdissection of Cells and Isolation of High-Quality RNA After Cryosectioning. In: Jin, H., Kaloshian, I. (eds) RNA Abundance Analysis . Methods in Molecular Biology, vol 2170. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0743-5_3
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DOI: https://doi.org/10.1007/978-1-0716-0743-5_3
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