Fructans are linear or branched polymers containing a single sucrose
and repeating fructose residues. An early
model for fructan biosynthesis in higher plants suggested that partial
synthesis of the polymer occurred in the cell
cytosol. The current model suggests that synthesis requires the interaction
of two separate fructosyltransferases
located in the vacuole. Tobacco lines containing a chemically induced promoter,
directing expression of the
Bacillus amyloliquefaciens SacB gene in the present study, provided
an opportunity to regulate and target fructan
synthesis to the cytosol of transgenic plants. Induced expression of the gene
led to rapid destruction of leaf tissue.
Amino acid substitution at a highly conserved site (Arg331)
in the SacB gene reduced the fructosyltransferase
efficiency without reducing the invertase activity of the enzyme. Expression
of the mutant gene in transgenic
tobacco also resulted in leaf damage. However, the appearance of necrotic
tissue was greatly delayed. The results
suggest that the phenotype is due to accumulation of fructan in the cytosol.
Fructan metabolism in the cytosol of
potato tubers was also detrimental to development. Tuber size and starch
synthesis was significantly reduced in
lines containing the untargeted gene. Transgenic tobacco and potato
containing the SacB gene offer an
opportunity to study the metabolism of fructan and the effect of
accumulation on plant cell development.