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
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 wordsAuxin Cytokinin Root-knot nematodes Cyst nematodes Lateral root Infection assay
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.
- 3.McCarter JP (2008) Molecular approaches toward resistance to plant-parasitic nematodes. In: Berg RH, Taylor CG (eds) Cell biology of plant nematode parasitism. Springer, Berlin, pp 239–267Google Scholar
- 4.Bohlmann H (2015) Introductory chapter on the basic biology of cyst nematodes. In: Escobar C, Fenoll C (eds) Advances in botanical research, vol 73. Elsevier Academic Press, Oxford, pp 33–59Google Scholar
- 5.Escobar C, Barcala M, Cabrera J, Fenoll C (2015) Overview of root-knot nematodes and giant cells. In: Escobar C, Fenoll C (eds) Advances in botanical research, vol 73. Elsevier Academic Press, Oxford, pp 1–32Google Scholar
- 7.Cabrera J, Diaz-Manzano FE, Sanchez M, Rosso MN, Melillo T, Goh T et al (2014) A role for LATERAL ORGAN BOUNDARIES-DOMAIN 16 during the interaction Arabidopsis-Meloidogyne spp. provides a molecular link between lateral root and root-knot nematode feeding site development. New Phytol 203:632–645CrossRefPubMedGoogle Scholar
- 18.Cabrera J, Díaz-Manzano FE, Barcala M, Arganda-Carreras I, de Almeida-Engler J, Engler G et al (2015) Phenotyping nematode feeding sites: three dimensional reconstruction and volumetric measurements of giant cells induced by root-knot nematodes in Arabidopsis. New Phytol 206:868–880CrossRefPubMedGoogle Scholar