Cytosolic expression of the Bacillus amyloliquefaciens SacB protein inhibits tissue development in transgenic tobacco and potato

Abstract

Fructans are linear or brancbed polymers containing a single sucrose and repeating fructose residues. An early model for fructan biosyntbesis in bigber plants suggested that partial synthesis of tbe polymer occurred in tbe cell cytosol. Tbe current model suggests tbat 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 tbe present study, provided an opportunity to regulate and target fructan synthesis to tbe cytosol of transgenic plants. Induced expression of tbe gene led to rapid destruction of leaf tissue. Amino acid substitution at a bigbly conserved site (Arg^^^) in tbe SacB gene reduced tbe fructosyltransferase efficiency witbout reducing tbe invertase activity of tbe enzyme. Expression of tbe mutant gene in transgenic tobacco also resulted in leaf damage. Hov^^ever, the appearance of necrotic tissue was greatly delayed. The results suggest tbat tbe pbenotype is due to accumulation of fructan in tbe cytosol. Fructan metabolism in tbe cytosol of potato tubers was also detrimental to development. Tuber size and starcb synthesis was significantly reduced in lines containing tbe untargeted gene. Transgenic tobacco and potato containing tbe SacB gene offer an opportunity to study tbe metabolism of fructan and the effect of accumulation on plant cell development.

Fructan and its relationship to abiotic stress tolerance in plants

“…In many cases, adverse effects of transgene expression on host metabolism or development, such as growth retardation, necrotic lesions, or sterility, were reported (e.g., [ 164 ]). This, as well as low concentrations of bacterial fructans, raised concerns about the physiological significance of these effects on stress tolerance [ 162 , 165 ].…”

Herbicide safeners: a review

“…Subsequent treatment of transgenic plants with 2-CBSU enabled the regulation of fructan synthesis in the cytosol and, thus, the study of the effect of fructan accumulation on plant development. 170…”