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JXB Advance Access originally published online on July 16, 2004
Journal of Experimental Botany 2004 55(403):1643-1654; doi:10.1093/jxb/erh195
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Journal of Experimental Botany, Vol. 55, No. 403, © Society for Experimental Biology 2004; all rights reserved

RESEARCH PAPER

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

Claudia Tomiko Otsu1,2 *, Idalete daSilva1,3 *, Jeanne Blanco de Molfetta1, Lara Regina da Silva1, Janice de Almeida-Engler4 {dagger}, Gilbert Engler4 {dagger}, Paulo Cesar Torraca1, Gustavo Henrique Goldman5 and Maria Helena S. Goldman1,{ddagger}

1Depto. Biologia FFCLRP/Universidade de São Paulo, Av. Bandeirantes, 3900 Ribeirão Preto, SP 14040-901, Brazil
2Depto. Genética FMRP/ Universidade de São Paulo, Av. Bandeirantes, 3900 Ribeirão Preto, SP 14040-900, Brazil
3FCAV/Universidade Estadual Paulista, Rod. Prof. Paulo Donato Castellane, s/no. Jaboticabal, SP 14884-900, Brazil
4Department of Molecular Genetics, Flanders Interuniversity Institute for Biotechnology (VIB), Ghent University, K.L. Ledeganckstraat 35, Gent B-9000, Belgium
5Depto. C. Farmacêuticas FCFRP/Universidade de São Paulo, Av. do Café s/no. Ribeirão Preto, SP 14040-903, Brazil

{ddagger} To whom correspondence should be addressed. Fax: +55 16 633 1758. E-mail: mgoldman{at}ffclrp.usp.br

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.

Key words: ABC transporter, developmentally regulated, stamen expression, stigma-predominant expression, stigmatic secretory zone, White subfamily


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