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JXB Advance Access originally published online on April 8, 2004
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Journal of Experimental Botany, Vol. 55, No. 399, pp. 983-992, May 1, 2004
© 2004 Oxford University Press

Cell and Molecular Biology, Biochemistry and Molecular Physiology

RNA interference in Agrobacterium rhizogenes-transformed roots of Arabidopsis and Medicago truncatula

Received 31 October 2003; Accepted 2 February 2004

Erik Limpens, Javier Ramos, Carolien Franken, Vered Raz, Bert Compaan, Henk Franssen, Ton Bisseling and René Geurts*

Laboratory of Molecular Biology, Wageningen University, Dreijenlaan 3, 6703HA Wageningen, The Netherlands

* To whom correspondence should be addressed. Fax: +31 (0)317 483584. E-mail: rene.geurts{at}wur.nl

RNA interference (RNAi) is a powerful reverse genetic tool to study gene function. The data presented here show that Agrobacterium rhizogenes-mediated RNAi is a fast and effective tool to study genes involved in root biology. The Arabidopsis gene KOJAK, involved in root hair development, was efficiently knocked down. A. rhizogenes-mediated root transformation is a fast method to generate adventitious, genetically transformed roots. In order to select for co-transformed roots a binary vector was developed that enables selection based on DsRED1 expression, with the additional benefit that chimaeric roots can be discriminated. The identification of chimaeric roots provided the opportunity to examine the extent of systemic spread of the silencing signal in the composite plants of both Arabidopsis and Medicago truncatula. It is shown that RNA silencing does not spread systemically to non-co-transformed (lateral) roots and only inefficiently to the non-transgenic shoot. Furthermore, evidence is presented which shows that RNAi is cell autonomous in the root epidermis.

Key words: A. rhizogenes, Arabidopsis, Medicago truncatula, RNAi, roots, silencing, systemic spreading.


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