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JXB Advance Access originally published online on August 8, 2003
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Journal of Experimental Botany, Vol. 54, No. 391, pp. 2245-2250, October 1, 2003
© 2003 Oxford University Press

A method for the isolation of root hairs from the model legume Medicago truncatula

Received 12 December 2002; Accepted 13 June 2003

Javier Ramos*,1,2 and Ton Bisseling2

1 Departamento de Nutrición Vegetal, Estación Experimental de Aula Dei, Consejo Superior de Investigaciones Científicas, Apdo 202, 50080 Zaragoza, Spain
2 Department of Molecular Biology, University of Wageningen, Dreijenlaan 3, 6703 HA Wageningen, The Netherlands

* To whom correspondence should be addressed. Fax: +34 976 716145. E-mail: jramos@ eead.csic.es

A new method for the isolation of root hairs from the model legume, Medicago truncatula, was developed. The procedure involves the propagation of detached roots on agar plates and the collection of root hairs by immersion in liquid nitrogen. Yields of up to 40 µg of root hair protein were obtained from 50–100 root tips grown for 3 weeks on a single plate. The high purity of the root hair fraction was monitored by western blot analysis using an antibody to the pea epidermis specific protein PsRH2. Sequence analyses revealed that the protein homologous to PsRH2 in M. truncatula, MtRH2, is identical to the root protein MtPR10-1. The MtRH2 protein proved to be a useful endogenous marker to monitor root hair isolation since it is also specifically expressed in the root epidermis.

Key words: Green fluorescent protein (GFP), legume, Medicago truncatula, PR10 defence protein, root hairs.


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