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First published online January 9, 2008; 10.1104/pp.107.109371

Plant Physiology 146:1128-1141 (2008)
© 2008 American Society of Plant Biologists

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DEVELOPMENT AND HORMONE ACTION

Functional Characterization of PaLAX1, a Putative Auxin Permease, in Heterologous Plant Systems1,[W],[OA]

Klára Hoyerová2, Lucie Perry2,*, Paul Hand, Martina Lanková, Tomás Kocábek, Sean May3, Jana Kottová, Jan Paces, Richard Napier and Eva Zazímalová

Institute of Experimental Botany (K.H., L.P., M.L., J.K., E.Z.) and Institute of Molecular Genetics (J.P.), Academy of Sciences of the Czech Republic, CZ–142 20 Prague 4, Czech Republic; HRI/University of Warwick, Wellesbourne, Warwick CV35 9EF, United Kingdom (P.H., S.M., R.N.); Biological Centre of the Academy of Sciences of the Czech Republic, v.v.i., Institute of Plant Molecular Biology, CZ–370 05 Ceské Budejovice, Czech Republic (T.K.); and Department of Biochemistry (M.L.) and Department of Plant Physiology (E.Z.), Faculty of Science, Charles University, CZ–128 00 Prague 2, Czech Republic

We have isolated the cDNA of the gene PaLAX1 from a wild cherry tree (Prunus avium). The gene and its product are highly similar in sequences to both the cDNAs and the corresponding protein products of AUX/LAX-type genes, coding for putative auxin influx carriers. We have prepared and characterized transformed Nicotiana tabacum and Arabidopsis thaliana plants carrying the gene PaLAX1. We have proved that constitutive overexpression of PaLAX1 is accompanied by changes in the content and distribution of free indole-3-acetic acid, the major endogenous auxin. The increase in free indole-3-acetic acid content in transgenic plants resulted in various phenotype changes, typical for the auxin-overproducing plants. The uptake of synthetic auxin, 2,4-dichlorophenoxyacetic acid, was 3 times higher in transgenic lines compared to the wild-type lines and the treatment with the auxin uptake inhibitor 1-naphthoxyacetic acid reverted the changes caused by the expression of PaLAX1. Moreover, the agravitropic response could be restored by expression of PaLAX1 in the mutant aux1 plants, which are deficient in auxin influx carrier activity. Based on our data, we have concluded that the product of the gene PaLAX1 promotes the uptake of auxin into cells, and, as a putative auxin influx carrier, it affects the content and distribution of free endogenous auxin in transgenic plants.


1 This work was supported by the Grant Agency of the Czech Republic (project nos. 206/02/P106 and 206/02/0967), the Grant Agency of the Academy of Sciences of the Czech Republic (project nos. B6038203, A6038303, and KJB600380702), the Ministry of Education, Youth and Sports (project nos. LN06034 and 1M0520), institutional supports (AV0Z50380511 and AV0Z50510513), and the Biotechnology and Biological Sciences Research Council.

2 These authors contributed equally to the article.

3 Present address: Nottingham Arabidopsis Stock Centre, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicestershire LE12 5RD, UK.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Lucie Perry (perry{at}ueb.cas.cz).

[W] The online version of this article contains Web-only data.

[OA] Open Access articles can be viewed online without a subscription.

www.plantphysiol.org/cgi/doi/10.1104/pp.107.109371

* Corresponding author; e-mail perry{at}ueb.cas.cz.

Received September 18, 2007; accepted December 21, 2007; published January 9, 2008.




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