JXB Advance Access originally published online on January 24, 2005
Journal of Experimental Botany 2005 56(413):799-806; doi:10.1093/jxb/eri074
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RESEARCH PAPER |
The promoter of the leghaemoglobin gene VfLb29: functional analysis and identification of modules necessary for its activation in the infected cells of root nodules and in the arbuscule-containing cells of mycorrhizal roots
1Universität Bielefeld, Fakultät für Biologie, Lehrstuhl für Genetik, PO Box 100131, D-33501 Bielefeld, Germany
2International NRW Graduate School in Bioinformatics and Genome Research, Center for Biotechnology, Universität Bielefeld, D-33594 Bielefeld, Germany
To whom correspondence should be addressed. Fax: +49 (0)521 1065626. E-mail: Helge.Kuester{at}Genetik.Uni-Bielefeld.DE
In this study the further characterization of the Vicia faba leghaemoglobin promoter pVfLb29 is presented that was previously shown to be specifically active in the infected cells of root nodules and in arbuscule-containing cells of mycorrhizal roots. Using promoter studies in transgenic hairy roots of the Pisum sativum mutant RisNod24, disabled in the formation of functional arbuscules, VfLb29 promoter activity is assigned to later stages of arbuscule development. In order to narrow down the regions containing cis-acting elements of pVfLb29, the activity of five VfLb29 promoter deletions (–797/–31 to –175/–31 in relation to the start codon) fused to the gusAint coding region were tested in transgenic V. hirsuta hairy roots. The results specify a promoter region ranging from position –410 to –326 (85 bp) as necessary for gus expression in arbuscule-containing cells, whereas this segment is not involved in the nodule-specific activity. Sequence analysis of the pVfLb29 fragment –410/–326 (85 bp) revealed sequence motifs previously shown to be cis-acting elements of diverse promoters. To investigate the autonomous function of pVfLb29 regions for activation in arbuscule-containing cells, different regions of pVfLb29 from positions –410 to –198 were used to prepare chimeric promoter constructs for trans-activation studies. These fragments alone did not activate the mycorrhiza inactive promoter of the Vicia faba leghaemoglobin gene VfLb3, showing that the activation of pVfLb29 in arbuscule-containing cells is governed by a complex regulatory system that requires at least two modules located between position –410 and –31 of the VfLb29 gene.
Key words: Arbuscules, leghaemoglobin promoter, mycorrhizal roots, root nodules, Vicia faba
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