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We have used top-down metabolic control analysis to investigate the control of carbon flux through potato (Solanum tuberosum) plants during tuberisation. The metabolism of the potato plant was divided into two blocks of reactions (the source and sink blocks) that communicate through the leaf apoplastic sucrose pool. Flux was measured as the transfer of 14C from CO2 to the tuber. Flux and apoplastic sucrose concentration were varied either by changing the light intensity or using transgenic manipulations that specifically affect the source or sink blocks, and elasticity coefficients were measured. We have provided evidence in support of our assumption that apoplastic sucrose is the only communicating metabolite between the source and sink blocks. The elasticity coefficients were used to calculate the flux control coefficients of the source and sink blocks, which were 0.8 and 0.2, respectively. This work suggests that the best strategy for the manipulation of tuber yield in potato will involve increases in photosynthetic capacity, rather than sink metabolism.

Potato tubers are modified stems that have differentiated into storage organs. Factors such as day-length, nitrogen supply, and levels of the phytohormones cytokinin and gibberellic acid, are known to control tuberization. Morphological changes during tuber initiation are accompanied by the accumulation of a characteristic set of proteins, thought to be involved in N-storage (i.e. patatin) or defense against microbial or insect attack (i.e. proteinase inhibitor II). Additionally, deposition of large amounts of starch occurs during tuber formation, which is paralleled by an increase in sucrose synthase and other enzymes involved in starch biosynthesis (i.e. ADP-glucose pyrophosphorylase, starch synthases, and branching enzyme). Potential controlling mechanisms for genes expressed during tuberization are discussed.

A chimaeric antisense construct was used to reduce the activities of the two major starch-synthase isoforms in potato tubers simultaneously. A range of reductions in total starch-synthase activities were found in the resulting transgenic plants, up to a maximum of 90% inhibition. The reduction in starch-synthase activity had a profound effect on the starch granules, which became extremely distorted in appearance compared with the control lines. Analysis of the starch indicated that the amounts produced in the tubers, and the amylose content of the starch, were not affected by the reduction in activity. In order to understand why the starch granules were distorted, amylopectin was isolated and the constituent chain lengths analysed. This indicated that the amylopectin was very different to that of the control. It contained more chains of fewer than 15 glucose units in length, and fewer of between 15 and 80 glucose units. In addition, the amylopectin contained more very long chains. Amylopectin from plants repressed in just one of the activities of the two starch-synthase isoforms, which we have reported upon previously, were also analysed. Using a technique different to that used previously we show that both isoforms also affect the amylopectin, but in a way that is different to when both isoforms are repressed together.

The Plant Research Department integrates modern post-genomic tools to improve our understanding of plants. The aim is to develop crops with improved agronomic traits and to engineer high-value plants, which are able to meet the growth conditions of thefuture environment. The department is divided into six research programmes that are linked through their individual expertise delivered to the rest of the department. Three programmes are engaged in improving the agronomic performance of plants. Genetictools are being developed to enhance the nutrient efficiency of plants, to strengthen the ability of plants to resist fungal attack, and to optimise flowering time. Two programmes are devoted to improving the market value of plant products. Plants withenhanced nutritional value, or that contain novel renewable resources, are designed to add value to the European Agro-Industries. A sixth programme studies the effects of the future climate on plant growth, and the performance of newly designed crops andtheir interaction with the environment. Several activities in the area of Functional Genomics integrate the department. Each programme contains special expertise in the fields of genome, transcriptome, proteome, and metabolome analyses, which aredelivered throughout the department. The Plant Research Department is unique in that these activities are supplemented with a broad expertise in environmental analysis, allowing the interpretation of large biological data sets in the context of factorsaffecting plant growth.

Während der Entwicklung von Früchten der Tomate (Sorte Micro-Tom) wurde der Kohlenhydrat-Stoffwechsel untersucht. Es wurde ein Unterschied zwischen dem Metabolismus im Perikarp und dem des Plazenta-Gewebes gefunden. Stärke wurde in der Plazenta langsamer abgebaut als im Perikarp, während lösliche Zucker im Perikarp stärker akkumulierten. Die Aktivitäten der glykolytischen Enzyme tendierten zu einem Maximum 40 Tage nach der Blüte. Weiterhin wurde die Expression einiger plastidärer Transporter untersucht. Sowohl der Triosephosphat-Tranporter (TPT) als auch der Glucose-6-phosphat-Transporter wurden am stärksten in grünen Früchten exprimiert, während der Reife nahm die Expression ab. Der ATP/ADP-Transporter wurde während der Fruchtentwicklung nur schwach exprimiert.Es besteht die Hypothese, daß die Rolle der drei Enzyme plastidäre Fructose-1,6-Bisphosphatase (cp-FBPase), ADP-Glucose Pyrophosphorylase (AGPase) und Glucan Wasser Dikinase (GWD) darin besteht, die Stärke-Akkumulation in der frühen Entwicklung der Tomaten-Frucht zu beeinflussen. Diese Hypothese wurde unter Verwendung der Antisense-Technik für die plastidären FBPase (unter der Kontrolle des B33 Promoters), sowie für die AGPase und die GWD (beide unter der Kontrolle des CaMV 35S-Promoters) in der Tomaten-Kultivar Moneymaker untersucht. Die Repression von plastidärer FBPase oder AGPase in der Frucht der Tomate scheint die Metaboliten-Konzentrationen nicht so stark wie in den Blättern zu beeinflussen. Der Grund hierfür ist wahrscheinlich, daß jede Veränderung durch die Fähigkeit der Frucht, Zucker zu importieren, abgepuffert wird. Auf der anderen Seite hatte die Repression des GWD Proteins in der Frucht der Tomate starke Effekte auf die Metaboliten-Konzentrationen. Carbohydrate metabolism was studied during the development of fruits of the tomato cultivar Micro-Tom. The metabolism of the pericarp and placental tissues was found to be different. Starch being degraded more slowly in the placenta than in the pericarp, while soluble sugars accumulated to a greater extent in the pericarp. The activities of glycolytic enzymes tended to peak at 40 days after flowering. The expression of some plastidial transporters was also studied. Both the triose phosphate transporter (TPT) and Glucose-6-Phosphate (Glc-6-P) transporter were expressed greatest in green fruits, before declining. The expression of the triose phosphate transporter (TPT) was greater than that of Glc-6-P transporter. The ATP/ADP transporter was expressed to a low level throughout fruit development. The role of three enzymes Chloroplastic Fructose-1,6-bisphosphatase (cp-FBPase), ADP-glucose Pyrophosphorylase (AGPase) and Glucan Water Dikinase (GWD) protein are thought to influence the accumulation of starch in early development in tomato fruit were studied in normal sized tomatoes of the cultivar Moneymaker using antisense technique under the control of the patatin B33 promoter in the case of cp-FBPase, and the CaMV 35S promoter in the case of AGPase and GWD protein. It appears that repression of cp-FBPase and AGPase in tomato fruits does not influence metabolite levels as greatly as it does in leaves, possibly because any alterations are buffered by the ability of the fruit to import sugars. On the other hand, the repression of GWD protein in tomato fruits has a strong effect on metabolite levels.