Molecular Biology and Genetics

Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 7 No. 1, Issue of April 15, 2004
© 2004 by Pontificia Universidad Católica de Valparaíso -- Chile Received September 28, 2003 / Accepted February 25, 2004
RESEARCH ARTICLE

Isolation and study of a ubiquitously expressed tomato pectin methylesterase regulatory region

Martín-Ernesto Tiznado-Hernández*
Departamento de Tecnología de Alimentos de Origen Vegetal
Centro de Investigación en Alimentación y Desarrollo, A.C.
Carretera a la Victoria km. 0.6
Apartado Postal 1735
Hermosillo, Sonora, 83000, México
Tel: 52 662 80 00 55
Fax: 52 662 280 04 22
E-mail: tiznado@cascabel.ciad.mx

Joel Gaffe
Genetique Moleculaire des Plantes
Universite Joseph Fourier
Cermo BP 53. 38041 Grenoble Cedex 9, France
Fax: 33 4 76 51 43 36
Tel: 33 4 76 51 44 41
E-mail: joel.gaffe@ujf-grenoble.fr

Avtar K. Handa
Department of Horticulture and Landscape Architecture
PURDUE University
1165 Horticulture Building
West Lafayette, IN, 47907-1165, USA
Tel: 765 494-1339
Fax: 765 494-0391
E-mail: handa@hort.purdue.edu

*Corresponding author


Keywords: pectin Methylesterase, promoter analysis, tobacco transgenic plants, tomato.

Abstract
Full Text

Pectin methylesterase (PME) is an enzyme located in the plant cell wall of higher plants whose physiological role is largely unknown. We had isolated a PME gene from a tomato genomic library, including 2.59 kb of 5´ flanking region and the coding region. Both coding and promoter region were sequenced and computer analyzed. Tobacco transgenic plants were created harboring constructs in which 2.596 Kb, 1.306 Kb and 0.267 Kb sizes of the promoter were driving the expression of β-Glucuronidase gene (GUS). GUS activity was studied by histochemical and fluorometric assays. Two introns of 106 and 1039 bp were found in the coding region and phylogenetic analysis placed this PME gene closer to genes from Citrus sinensis and Arabidopsis thaliana than tomato fruit-specific PME genes. In the promoter, it was found direct repeats, perfect inverted repeats and light responsive elements. GUS histochemical analysis showed activity in all plant tissues with the exception of pollen. The reduction in the promoter size induced a reduction in GUS activity in root, stem and leaf. Furthermore, root and leaf showed the highest and lowest activity, respectively. We had isolated a tomato PME gene with novel characteristics as compared with other known PME genes from tomato.

 
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