In vivo expression of a Cicer arietinum beta-galactosidase in potato tubers leads to a reduction of the galactan side-chains in cell wall pectin.
We report the generation of Solanum tuberosum transformants expressing Cicer arietinum betaIII-Gal. betaIII-Gal is a beta-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 beta-galactosidase cDNAs, and named CanBGal-3. CanBGal-3 cDNA was expressed in potato under the control of the granule-bound starch synthase promoter. Three betaIII-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, beta-galactanase and beta-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-beta-galactan side-chains, as revealed by analysis with LM5 specific antibodies. Our results confirm the notion that the pectin-degrading activity of chickpea betaIII-Gal reported in vitro also occurs in vivo and in other plants, and confirm the involvement of betaIII-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.
Martín
Ignacio
I
Departamento de Fisiología Vegetal, Centro Hispano Luso de Investigaciones Agrarias, Universidad de Salamanca, Plaza Doctores de la Reina s/n, Campus Miguel Unamuno, Spain.
Dopico
Berta
B
Muñoz
Francisco J
FJ
Esteban
Rocío
R
Oomen
Ronald J F J
RJ
Driouich
Azeddine
A
Vincken
Jean-Paul
JP
Visser
Richard
R
Labrador
Emilia
E
D015152:Blotting, Northern; D002473:Cell Wall / Q000378:metabolism*; D029863:Cicer / Q000201:enzymology*; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D005685:Galactans / Q000737:chemistry*; D010368:Pectins / Q000737:chemistry / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D011198:Solanum tuberosum / Q000235:genetics*; D017550:Spectroscopy, Fourier Transform Infrared; D001616:beta-Galactosidase / Q000235:genetics*
D018076:DNA, Complementary; D005685:Galactans; D012333:RNA, Messenger; D010368:Pectins; D001616:beta-Galactosidase