SummaryTo analyse carbohydrate metabolism and its role during early seed development of barley we characterised genes encoding two cell wall-bound invertases (HvCWINV1 and HvCWINV2) and two putative hexose transporter-like genes (HvSTP1 and HvSTP2). No typical vacuolar invertase gene could be identified. Instead, a gene encoding sucrose:fructan 6-fructosyltransferase (HvSF6FT1), an enzyme with soluble acid invertase activity, was isolated and characterised. Furthermore, enzyme activities and sugar levels were measured. HvSF6FT1-mRNA levels and acid soluble invertase activity are highest in the maternal pericarp 1-2 days after flowering (DAF). HvSF6FT1 is strongly expressed in regions flanking the main vascular bundle and to a lower extent in filial endospermal transfer cells, which persist until maturity and never accumulate starch. In contrast, cell wall-bound invertase HvCWINV2 is expressed early in development mainly in the style region and later on in pericarp areas which transiently accumulate starch and undergo degradation later in development. The hexose transporter HvSTP2 shows a temporal and spatial expression pattern similar to HvCWINV2. Transcripts of HvCWINV1 have been localised within the first row of endospermal cells and in the outermost area of the nucellar projection as well as in endospermal transfer cells before starch filling; the same regions of the endosperm are labelled with a hexose transporter HvSTP1-probe. HvSTP1 is expressed at very low levels within the pericarp but much higher in the syncytial endosperm at 3 DAF and in endospermal transfer cells 7 DAF. The temporal and spatial association of HvCWINV1 and HvSTP1 expression indicates that hexoses liberated by the invertase within the endospermal cavity are taken up by the transporter to be delivered into the central uncellularised space of the endosperm to supply mitotically active endosperm cells with hexoses. The results are discussed and compared with published data on the role of soluble sugars as signal molecules in seed developmental processes.

Summary Different aspects of barley grain development have been studied in detail, but a more global analysis of gene expression patterns is still missing. We have employed macro arrays, containing 1184 unique sequences from 1421 barley cDNA fragments, to study gene expression profiles in maternal and filial tissues of developing barley caryopses from fertilization to early storage phase. Principle component analysis (PCA) defined distinct expression networks in the pre‐storage (0, 2, and 4 days after flowering (DAF)) and early storage phase (10 and 12 DAF). During an intermediate phase (6 and 8 DAF), PCA visualizes a dramatic re‐programming of the transcriptional machinery. In maternal tissues, a large set of protein‐mobilizing enzyme mRNAs, together with upregulated lipid‐mobilizing enzyme and downregulated reactive oxygen species (ROS)‐scavenging enzyme genes, suggests mobilization of stored compounds and programmed cell death (PCD). In the filial tissue fraction, a set of genes highly expressed during the pre‐storage phase is involved in growth processes, including cell wall biosynthesis. The data suggest that the necessary UDP‐glucose is provided both by sucrose synthase (isoform 3) and an invertase‐driven pathway. Further, major developmental changes in pathways producing energy are predicted. A bell‐shaped expression profile with a peak during the intermediate phase is characteristic for genes associated with photosynthesis and ATP production. The photosynthesis‐determined increase of ATP concentration could be a prerequisite for the initiation of grain filling, dominated by starch and storage protein synthesis. Storage product accumulation is accompanied by high transcriptional activity of genes involved in glycolysis and fermentation, as well as in the citric acid cycle.

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SummaryThe small subunit of a Vicia faba ADP-glucose pyrophosphorylase (AGP) cDNA was expressed in antisense orientation in Vicia narbonensis under the control of the seed-speci®c legumin B4 promoter. From several independent transgenic lines both ADP-glucose pyrophosphorylase AGP-mRNA and AGP enzyme activity were reduced by up to 95% in the cotyledons during the mid-to late-maturation phase. Starch was moderately decreased and sucrose was increased. In two of three lines, transcripts encoding the large subunit of AGP and the storage protein vicilin were increased, whereas legumin B-mRNA was decreased. Transcripts of other storage-associated genes were not altered. The cotyledons contained more protein and total nitrogen. Despite the reduction in starch, total carbon was not decreased and dry weight was unchanged. Compared to the wild type, transgenic seeds contained more water and accumulated dry weight during a longer period, and therefore had a prolonged seed-®lling period. Transgenic cotyledon cells of comparable age to the wild type were more highly vacuolated and contained smaller starch grains, indicating a delay in cellular differentiation. We conclude that a speci®c alteration in carbon metabolism can have pleiotropic effects on water and nitrogen content and induces temporal changes in seed development.

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