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Plant and Cell Physiology Advance Access originally published online on August 2, 2005
Plant and Cell Physiology 2005 46(10):1613-1622; doi:10.1093/pcp/pci177
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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 Martín1, Berta Dopico1, Francisco J. Muñoz1, Rocío Esteban1, Ronald J. F. J. Oomen2, Azeddine Driouich3, Jean-Paul Vincken2, Richard Visser2 and Emilia Labrador1,*

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 Aignan, Cedex, France

* Corresponding author: E-mail, labrador{at}usal.es; Fax, +34-923-294682.

We report the generation of Solanum tuberosum transformants expressing Cicer arietinum ßIII-Gal. ßIII-Gal is a ß-galactosidase able to degrade cell wall pectins during cell wall loosening that occurs prior to cell elongation. cDNA corresponding to the gene encoding this protein was identified among several chickpea ß-galactosidase cDNAs, and named CanBGal-3. 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 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% 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 analysis with LM5 specific antibodies. Our results confirm the notion that the pectin-degrading activity of chickpea ßIII-Gal reported in vitro also occurs in vivo and in other plants, and confirm the involvement of ßIII-Gal in the cell wall autolysis process. An increase in the homogalacturonan content of transgenic tuber cell walls was also observed by Fourier transform infrared spectroscopy (FTIR) analysis.

(Received December 21, 2004; Accepted July 22, 2005)
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