Temporal and spatial expression of hexose transporters in developing tomato (Lycopersicon esculentum) fruit.

Correlative physiological evidence suggests that membrane transport into storage parenchyma cells is a key step in determining hexose levels accumulated in tomato (Lycopersicon esculentum Mill.) fruit (Ruan et al. 1997). Expression of three previously identified hexose transporter genes (LeHT1, 2 and 3) demonstrated that LeHT3, and to a lesser extent LeHT1, are the predominant transporters expressed in young fruit (10 d after anthesis; DAA). Expression of both transporters dropped sharply until 24 DAA, after which only LeHT3 expression remained at detectable levels through to fruit ripening. LeHT2 was not expressed substantially until the onset of fruit ripening. For fruit at both 10 and 30 DAA, LeHT3 transcripts were detected in storage parenchyma cells of the outer pericarp tissue, but not in vascular bundles or the first layer of parenchyma cells surrounding these bundles. In contrast to LeHT gene expression, hexose transporter protein levels were maximal between 20 and 30 DAA, which corresponded to the period of highest hexose accumulation. The delayed appearance of transporter protein is consistent with some form of post-transcriptional regulation. Based on these analyses, LeHT3 appears to be responsible for the rapid hexose accumulation in developing tomato fruit.

Increased capacity for sucrose uptake leads to earlier onset of protein accumulation in developing pea seeds.

Pea (Pisum sativum L.) cotyledons, overexpressing a potato sucrose transporter (StSUT1), were used to explore the hypothesis that sucrose stimulates the onset of storage protein biosynthesis. The study focused on the transition between pre-storage and storage phases of seed development. During this period supply of sucrose and hexose to transgenic cotyledons was unaffected by StSUT1 expression. However, protoplasmic levels of sucrose but not hexoses were elevated in transgenic cotyledons. Total protein levels in cotyledons followed the same temporal trend as observed for sucrose and this was reflected in an earlier appearance of protein bodies. Protein levels in wild type and StSUT1 cotyledons were found to lie on the same sucrose dose-response curve and this could be reproduced in vitro when wild type cotyledons were cultured on media containing various sucrose concentrations. Rates of [14C]sucrose uptake and incorporation into polymeric forms were consistent with protoplasmic sucrose supplying a proportion of the carbon skeletons required for storage protein accumulation. In addition, vicilin gene expression was up-regulated earlier in StSUT1 cotyledons. We conclude that sucrose functions both as a signal and fuel to stimulate storage protein accumulation and assembly into protein bodies. An earlier stimulation of storage protein synthesis is considered to largely account for the 14% increase in protein levels of StSUT1 seeds at harvest.

Seed-specific overexpression of a potato sucrose transporter increases sucrose uptake and growth rates of developing pea cotyledons.

During the storage phase, cotyledons of developing pea seeds are nourished by nutrients released to the seed apoplasm by their maternal seed coats. Sucrose is transported into pea cotyledons by sucrose/H+ symport mediated by PsSUT1 and possibly other sucrose symporters. PsSUT1 is principally localised to plasma membranes of cotyledon epidermal and subepidermal transfer cells abutting the seed coat. We tested the hypothesis that endogenous sucrose/H+ symporter(s) regulate sucrose import into developing pea cotyledons. This was done by supplementing their transport activity with a potato sucrose symporter (StSUT1), selectively expressed in cotyledon storage parenchyma cells under control of a vicilin promoter. In segregating transgenic lines, enhanced [(14)C]sucrose influx into cotyledons above wild-type levels was found to be dependent on StSUT1 expression. The transgene significantly increased (approximately 2-fold) transport activity of cotyledon storage parenchyma tissues where it was selectively expressed. In contrast, sucrose influx into whole cotyledons through the endogenous epidermal transfer cell pathway was increased by only 23% in cotyledons expressing the transgene. A similar response was found for rates of biomass gain by intact cotyledons and by excised cotyledons cultured on a sucrose medium. These observations demonstrate that transport activities of sucrose symporters influence cotyledon growth rates. The attenuated effect of StSUT1 overexpression on sucrose and dry matter fluxes by whole cotyledons is consistent with a large proportion of sucrose being taken up at the cotyledonary surface. This indicates that the cellular location of sucrose transporter activity plays a key role in determining rates of sucrose import into cotyledons.