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JXB Advance Access published online on July 16, 2004
Journal of Experimental Botany, doi:10.1093/jxb/erh195
© 2004 by Oxford University Press
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Received July 24, 2003
Accepted May 7, 2004

RESEARCH PAPER

NtWBC1, an ABC transporter gene specifically expressed in tobacco reproductive organs

Claudia Tomiko Otsu 1 *, Idalete daSilva 2 *, Jeanne Blanco de Molfetta 3, Lara Regina da Silva 3, Janice de Almeida-Engler 4, Gilbert Engler 4, Paulo Cesar Torraca 3, Gustavo Henrique Goldman 5, Maria Helena S. Goldman 3*

1 Depto. Biologia FFCLRP/Universidade de São Paulo, Av. Bandeirantes, 3900 Ribeirão Preto, SP 14040-901, Brazil; Depto. Genética FMRP/ Universidade de São Paulo, Av. Bandeirantes, 3900 Ribeirão Preto, SP 14040-900, Brazil
2 Depto. Biologia FFCLRP/Universidade de São Paulo, Av. Bandeirantes, 3900 Ribeirão Preto, SP 14040-901, Brazil; FCAV/Universidade Estadual Paulista, Rod. Prof. Paulo Donato Castellane, s/no. Jaboticabal, SP 14884-900, Brazil
3 Depto. Biologia FFCLRP/Universidade de São Paulo, Av. Bandeirantes, 3900 Ribeirão Preto, SP 14040-901, Brazil
4 Department of Molecular Genetics, Flanders Interuniversity Institute for Biotechnology (VIB), Ghent University, K.L. Ledeganckstraat 35, Gent B-9000, Belgium; Present address: UMR-Interactions Plantes-Microorganismes et Sante Vegetale, INRA/CNRS/UNSA, BP 167, 400 Route des Chappes, 06903 Sophia Antipolis Cedex, France
5 Depto. C. Farmacêuticas FCFRP/Universidade de São Paulo, Av. do Café s/no. Ribeirão Preto, SP 14040-903, Brazil

* To whom correspondence should be addressed. E-mail: mgoldman{at}ffclrp.usp.br.


  Abstract

To identify genes specifically or predominantly expressed in the stigmas/styles and to establish their possible function in the reproductive process of plants, a tobacco stigma/style cDNA library was constructed and differentially screened, resulting in the isolation of several cDNA clones. The molecular characterization of one of these clones is described here. After sequencing the cDNA and the isolated genomic clone, it was determined that the corresponding gene encodes a protein containing an ATP-binding cassette, characteristic of ABC transporters. This gene, designated as NtWBC1 (Nicotiana tabacum ABC transporter of the White-Brown Complex subfamily), encodes a protein that contains the typical structure of the ‘half-transporters’ of the White subfamily. To establish the spatial expression pattern of the NtWBC1 gene, northern blot and real-time RT-PCR analyses with total RNA from roots, stems, leaves, sepals, petals, stamens, stigmas/styles, ovaries, and seeds were performed. The result revealed a transcript of 2.5 kb present at high levels in stigmas and styles and a smaller transcript (2.3 kb) present at a lower level in stamens. NtWBC1 expression is developmentally regulated in stigmas/styles, with mRNA accumulation increasing toward anthesis. In situ hybridization experiments demonstrated that NtWBC1 is expressed in the stigmatic secretory zone and in anthers, at the stomium region and at the vascular bundle. NtWBC1 is the first ABC transporter gene with specific expression in plant reproductive organs to be identified and its expression pattern suggests important role(s) in the reproductive process.

Keywords: ABC transporter; developmentally regulated; stamen expression; stigma-predominant expression; stigmatic secretory zone; White subfamily.

The first and second authors have contributed equally to this work.


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