JXB Advance Access originally published online on April 2, 2007
Journal of Experimental Botany 2007 58(7):1717-1728; doi:10.1093/jxb/erm025
© 2007 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)
RESEARCH PAPER |
Expression and functional analysis of metal transporter genes in two contrasting ecotypes of the hyperaccumulator Thlaspi caerulescens
1Crop Performance and Improvement Division, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, UK
2Agriculture and Environment Division, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, UK
To whom correspondence should be addressed. E-mail: malcolm.hawkesford{at}bbsrc.ac.uk
Zinc (Zn) hyperaccumulation is a constitutive property of Thlaspi caerulescens, whereas cadmium (Cd) hyperaccumulation varies greatly among different ecotypes. The molecular basis of this variation is unknown. Ecotypic differences in the sequences and expression of four representative ZIP family transporter genes were investigated. Genome analysis indicated the presence of at least two closely related copies of the TcIRT1 gene in both Ganges (high Cd accumulating) and Prayon (low Cd accumulating) ecotypes, with different copies being expressed in each, and, furthermore, the two genes potentially encode different length transcripts. The predominant transcript in Prayon was truncated, missing sequence coding for the putative metal-binding site and the five C-terminal transmembrane helices. The two ecotypes were grown hydroponically ±Fe and Cd, and mRNA abundance determined for four ZIP genes. The four ZIP genes studied (TcIRT1, TcIRT2, TcZNT1, and TcZNT5) were expressed in roots only. TcIRT1 expression (full-length in Ganges, TcIRT1-1G; truncated in Prayon, TcIRT1-2P) was enhanced by Fe deficiency or by exposure to Cd. TcIRT2 expression was induced by Fe deficiency, but was unaffected by Cd exposure. TcZNT5-G showed greater expression in Prayon compared with Ganges. The functions of TcIRT1 from Ganges and Prayon and the Arabidopsis homologue were analysed by heterologous expression in yeast. All three IRT1 genes were able to facilitate growth on low Fe concentrations. Cd sensitivity of yeast was conferred in the order AtIRT1>TcIRT1-1G>TcIRT1-2P (truncated). Cd uptake after 4 h was only detectable following complementation by AtIRT1. The results suggest that although TcIRT1-G may be involved in Cd hyperaccumulation in the Ganges ecotype of T. caerulescens, the transporter expressed in yeast does not have an enhanced ability to transport Cd compared with AtIRT1. Therefore, the unique Cd-accumulating ability of the T. caerulescens Ganges ecotype must be due to the levels of expression of the protein or to other factors such as interacting proteins.
Key words: Cadmium, hyperaccumulation, IRT1, Thlaspi caerulescens, zinc, ZIP transporters
* Present address: Plant Biology, University of Fribourg, Fribourg, CH-1700, Switzerland.
Present address: Business and Innovation Unit, BBSRC, Swindon SN2 1UH, UK.
These authors contributed equally to this work.
Received 6 September 2006; Revised 10 January 2007 Accepted 24 January 2007
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