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Analysis of Expression Gradients of Developmental Regulators in Arabidopsis thaliana Roots

  • María Florencia Ercoli
  • Rodrigo Vena
  • Camila Goldy
  • Javier F. Palatnik
  • Ramiro E. Rodríguez
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1863)

Abstract

The regulatory mechanisms involved in plant development include many signals, some of them acting as graded positional cues regulating gene expression in a concentration-dependent manner. These regulatory molecules, that can be considered similar to animal morphogens, control cell behavior in developing organs. A suitable experimental approach to study expression gradients in plants is quantitative laser scanning confocal microscopy (LSCM) using Arabidopsis thaliana root tips as a model system. In this chapter, we outline a detailed method for image acquisition using LSCM, including detailed microscope settings and image analysis using FIJI as software platform.

Key words

miR396 PLETHORA Laser scanning confocal microscopy Root Arabidopsis thaliana 

Notes

Acknowledgments

We thank Philip Benfey for the AGL42-GFP reporter, Moritz Nowack for the pSMB-H2B-GFP reporter, Lieven de Veylder for the pGL2:H2B-YFP, pCO:H2B-YFP, and the pEND:H2B-YFP reporters, and, finally, Ben Scheres for the PLT1 and PLT2 transcriptional and translational reporters. We thank Carla Schommer for critical reading of the manuscript. MFE and CG were supported by fellowships from CONICET. RV is a technician of CONICET. JP and RER are members of CONICET and are supported by grants from ANPCyT.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • María Florencia Ercoli
    • 1
  • Rodrigo Vena
    • 1
  • Camila Goldy
    • 1
  • Javier F. Palatnik
    • 1
    • 2
  • Ramiro E. Rodríguez
    • 1
    • 2
  1. 1.IBR (Instituto de Biología Molecular y Celular de Rosario), CONICET, Universidad Nacional de RosarioRosarioArgentina
  2. 2.Centro de Estudios InterdisciplinariosUniversidad Nacional de RosarioRosarioArgentina

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