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Plant and Cell Physiology Advance Access published online on August 2, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci177
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received December 21, 2005
Accepted July 22, 2005

Regular Paper

In Vivo Expression of a Cicer arietinum ß-Galactosidase in Potato Tubers Leads to a Reduction of the Galactan Side-Chains in Cell Wall Pectin

Ignacio Martín 1, Berta Dopico 1, Francisco J. Muñoz 1, Rocío Esteban 1, Ronald J. F. J. Oomen 2, Azeddine Driouich 3, Jean-Paul Vincken 2, Richard Visser 2, and Emilia Labrador 1*

1 Departamento de Fisiología Vegetal, Centro Hispano Luso de Investigaciones Agrarias. Universidad de Salamanca. Plaza Doctores de la Reina s/n. Campus Miguel Unamuno. 37007 Salamanca, Spain
2 Wageningen University, Laboratory of Plant Breeding, Binnenhaven 5, 6709 PD Wageningen, The Netherlands
3 UMR CNRS 6037. IFRMP23. CCME. Université de Rouen. 76821 Mont Saint

* To whom correspondence should be addressed.
Emilia Labrador, E-mail: labrador{at}usal.es


  Abstract

We report the generation of Solanum tuberosum transformants expressing the Cicer arietinum ßIII-Gal. The ßIII-Gal is a ß-galactosidase able to degrade cell wall pectins during cell wall loosening that occurs prior to cell elongation. The cDNA corresponding to the gene encoding this protein was identified among several chickpea ß-galactosidase cDNAs, and was named CanBGal-3. The CanBGal-3 cDNA was expressed in potato under the control of the granule-bound starch synthase promoter. Three ßIII-Gal transformants with varying levels of expression were chosen for further analysis. The transgenic plants displayed no significant altered phenotype as compared to the wild type. However, ß-galactanase and ß-galactosidase activities were increased in the transgenic tuber cell walls and this affected the potato tuber pectins. A reduction in the galactosyl content of up to 50% as compared to the wild type was observed in the most extreme transformant, indicating a reduction of 1,4-ß-galactan side-chains, as revealed by the analysis with LM5 specific antibodies. Our results confirm the notion that the pectin-degrading activity of the chickpea ßIII-Gal reported in vitro also occurs in vivo and in other plants, and confirm the involvement of ßIII-Gal in the autolysis process. An increase in the homogalacturonan content of transgenic tuber cell walls was also observed by Fourier transform infrared spectroscopy (FTIR) analysis.

Keywords: Cell wall; chickpea; ß-galactosidase; pectin; potato; transgenic plant.
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