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JXB Advance Access first published online on April 18, 2005
This version published online on May 6, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri152
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved.
Received December 6, 2004
Accepted February 28, 2005

RESEARCH PAPER

Using a suppression subtractive library-based approach to identify tobacco genes regulated in response to short-term sulphur deficit

Anna Wawrzynska 1, Malgorzata Lewandowska 1, Malcolm J. Hawkesford 2, and Agnieszka Sirko 1*

1 Institute of Biochemistry and Biophysics, Polish Academy of Sciences, ul. Pawinskiego 5A, 02-106 Warsaw, Poland
2 Rothamsted Research, Harpenden AL5 2JQ, UK

* To whom correspondence should be addressed.
Agnieszka Sirko, E-mail: asirko{at}ibb.waw.pl


  Abstract

Monitoring expression at the transcriptional level is an essential first step for the functional analysis of plant genes. Genes encoding proteins directly involved in sulphur metabolism constitute only a small fraction of all the genes affected by sulphur deficiency stress. Transcriptional responses to various periods of sulphur deprivation have been extensively studied in Arabidopsis thaliana; however, no corresponding data are available for Solanaceae sp. To address this problem, a subtractive library-based approach to search for tobacco genes regulated by a short-term sulphur starvation has been adopted. In this work, 38 genes were identified, of which 22 were regulated positively and 16 were regulated negatively. The transcript levels of the representative genes were monitored in four parts of the plants (mature and immature leaves, stems, and roots), which exhibited differential sulphur deficiency. Interestingly, some genes exhibit different regulation of expression in different parts of the plants. Database analysis allowed assignment of the potential function for many of the identified genes; however, the functions of a small number of genes strongly regulated by sulphur starvation remain unknown. The genes were grouped into nine functional categories, each including both up- and down-regulated genes. The possible links between the identified regulated genes and sulphur metabolism are considered, and compared where possible with expression patterns in Arabidopsis thaliana. Although no obvious regulatory genes were identified, the genes encoding proteins of unknown function remain as potential components of the regulatory processes.

Keywords: Starvation; stress signaling; sulphur metabolism; suppression subtractive hybridization; tobacco.

This is a new version of this article as there were errors in the GenBank accession numbers in Table 1 in the first version.


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