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A Standardized Method to Assess Infection Rates of Root-Knot and Cyst Nematodes in Arabidopsis thaliana Mutants with Alterations in Root Development Related to Auxin and Cytokinin Signaling

  • Rocío Olmo
  • Ana Cláudia Silva
  • Fernando E. Díaz-Manzano
  • Javier Cabrera
  • Carmen Fenoll
  • Carolina Escobar
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1569)

Abstract

Plant parasitic nematodes cause a great impact in agricultural systems. The search for effective control methods is partly based on the understanding of underlying molecular mechanisms leading to the formation of nematode feeding sites. In this respect, crosstalk of hormones such as auxins and cytokinins (IAA, CK) between the plant and the nematode seems to be crucial. Thence, the study of loss of function or overexpressing lines with altered IAA and CK functioning is entailed. Those lines frequently show developmental defects in the number, position and/or length of the lateral roots what could generate a bias in the interpretation of the nematode infection parameters. Here we present a protocol to assess differences in nematode infectivity with the lowest interference of root architecture phenotypes in the results. Thus, tailored growth conditions and normalization parameters facilitate the standardized phenotyping of nematode infection.

Key words

Auxin Cytokinin Root-knot nematodes Cyst nematodes Lateral root Infection assay 

Notes

Acknowledgments

This work was supported by the Spanish Government (AGL2013-48787 to CE; and PCIN-2013-053 to CF) and by the Castilla-La Mancha Government (PEII-2014-020-P to CF). FEDM, RO, JC, and ACS acknowledge the European Social Fund, the University of Castilla La Mancha and the MINECO for cofinancing this work.

Supplementary material

332994_1_En_5_MOESM1_ESM.docx (603 kb)
Fig. S1 Plates within the growth chamber. (a) Plates with plants sown in a single row before covering them with the gauze; average light intensity received by the plants is 104 μmol/m2 s. (b) Plates covered with a gauze (plants) from fluorescent light exposure; average light intensity received by the plants is 48 μmol/m2 s. Scale bars: 10 cm (DOCX 602 kb)

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Rocío Olmo
    • 1
  • Ana Cláudia Silva
    • 1
  • Fernando E. Díaz-Manzano
    • 1
  • Javier Cabrera
    • 1
  • Carmen Fenoll
    • 1
  • Carolina Escobar
    • 1
  1. 1.Facultad de Ciencias Ambientales y BioquímicaUniversidad de Castilla-La ManchaToledoSpain

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