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Genetic and Phenotypic Analysis of Lateral Root Development in Arabidopsis thaliana

  • Selene Napsucialy-Mendivil
  • Joseph G. Dubrovsky
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1761)

Abstract

Root system formation to a great extent depends on lateral root (LR) formation. In Arabidopsis thaliana, LRs are initiated within a parent root in pericycle that is an external tissue of the stele. LR initiation takes place in a strictly acropetal pattern, whereas posterior lateral root primordium (LRP) formation is asynchronous. In this chapter, we focus on methods of genetic and phenotypic analysis of LR initiation, LRP morphogenesis, and LR emergence in Arabidopsis. We provide details on how to make cleared root preparations and how to identify the LRP stages. We also pay attention to the categorization of the LRP developmental stages and their variations and to the normalization of the number of LRs and LRPs formed, per length of the primary root, and per number of cells produced within a root. Hormonal misbalances and mutations affect LRP morphogenesis significantly, and the evaluation of LRP abnormalities is addressed as well. Finally, we deal with various molecular markers that can be used for genetic and phenotypic analyses of LR development.

Key words

Root branching Root system architecture Root primordium Lateral root Pericycle Root initiation Developmental stages Founder cells Quantitative analysis Morphogenesis 

Notes

Acknowledgments

We thank K. Nikoumanesh and H. H. Torres-Martínez for the critical reading of the manuscript, N. Doktor for the help with illustrations, and S. E. Ainsworth, J. M. Hurtado-Ramírez, and A. Ocádiz-Ramírez for their help with logistics. The work in the corresponding author laboratory is supported by the Mexican Consejo Nacional de Ciencia y Tecnología (CONACyT: Grant 237430) and Universidad Nacional Autónoma de México-DGAPA-PAPIIT (IN205315).

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Selene Napsucialy-Mendivil
    • 1
  • Joseph G. Dubrovsky
    • 1
  1. 1.Departamento de Biología Molecular de Plantas, Instituto de BiotecnologíaUniversidad Nacional Autónoma de México (UNAM)CuernavacaMexico

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