Down-regulation of a ripening-related beta-galactosidase gene (TBG1) in transgenic tomato fruits.
Exo-galactanase/beta-galactosidase (EC 3.2.1.23) activity is thought to be responsible for the loss of galactosyl residues from the cell walls of ripening tomatoes. Transgenic tomato plants (Lycopersicon esculentum Mill cv. Ailsa Craig) with reduced exo-galactanase/beta-galactosidase mRNA were generated to test this hypothesis and to investigate the role of the enzyme in fruit softening. A previously identified tomato beta-galactosidase cDNA clone, TBG1, was used in the experiments. Heterologous expression of the clone in yeast demonstrated that TBG1 could release galactosyl residues from tomato cell wall galactans. Transgenic plants showed a reduction in TBG1 mRNA to 10% of normal levels in the ripening fruits. However, despite the reduction in message, total beta-galactosidase and exo-galactanase activities were unaffected. Furthermore, there was no apparent effect on levels of cell wall galactosyl residues when compared with the control. It was concluded that during the ripening of tomato fruits a family of beta-galactosidases capable of degrading cell wall galactans are active and down-regulation of TBG1 message to 10% was insufficient to alter the degree of galactan degradation.
Carey
A T
AT
Department of Plant Genetics and Biotechnology, Horticulture Research International, Wellesbourne, Warwick CV35 9EF, UK.
Smith
D L
DL
Harrison
E
E
Bird
C R
CR
Gross
K C
KC
Seymour
G B
GB
Tucker
G A
GA
D001483:Base Sequence; D017931:DNA Primers; D015536:Down-Regulation; D018551:Solanum lycopersicum / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D011994:Recombinant Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics*
D017931:DNA Primers; D011994:Recombinant Proteins; D001616:beta-Galactosidase