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.
- infrared microspectroscopy
- natural substrate
Martin, I., Dopico, B., Munoz, F. J., Esteban, R., Oomen, R. J. F. J., Driouich, A., Vincken, J. P., Visser, R. G. F., & Labrador, E. (2005). In vivo expression of a Cicer arietinum B-galactosidase in potato tubers leads to a reduction of the Galactan side-chains in cell wall pectin. Plant and Cell Physiology, 46(10), 1613-1622. https://doi.org/10.1093/pcp/pci177