The aim of this proposal is to investigate factors controlling the chemical composition of tomato fruit, a crop of major economic importance worldwide. Both the flavour and nutritional quality of tomatoes are determined by the chemicals that accumulate during fruit ripening, yet we have only a limited understanding of how this process is controlled. In mature fruit, the cells are dominated by a compartment called the central vacuole, which contains most of the sap in fleshy fruit. This compartment can occupy as much as 95 % of the cell's volume, the remaining 5 % being taken up by the cell cytoplasm and outlying cell wall. As the tomato fruit grows, chemicals such as sugars, organic acids and amino acids are produced in the cytoplasm. They are then removed from their site of synthesis by transport into the central vacuole across the bounding membrane surrounding this compartment, called the tonoplast. But this traffic is not all one-way. As the fruit ripens, some solutes leave the vacuole to be re-metabolized in the cytoplasm, with other solutes moving back into the vacuole to compensate. Thus, the composition of the mature fruit is a complex outcome of metabolic events in the cytoplasm combined with transport of solutes across the tonoplast membrane. Whereas the pathways of basic metabolism in fruit cells are well understood, we have much less knowledge of the transport proteins that reside in the tonoplast membrane. In fact, we have indirect evidence that these proteins may play a much more important role in determining fruit composition than previously suspected. As the first part of this project, therefore, we shall isolate the tonoplast membrane from tomato fruit at defined stages during their development and analyse its protein content by mass spectrometry. This will provide a valuable inventory of proteins residing in the tonoplast membrane, and of their changes in abundance during the ripening process. By correlating these changes with the chemical composition of the fruit, we should obtain the first clues as to which tonoplast proteins are important in regulating transport across the vacuolar membrane. In another strand of the project, we will use a genetic approach to obtain independent information on factors controlling fruit composition. A powerful resource for this purpose is provided by the natural genetic variation found between cultivated tomatoes and their close relatives in the wild. Indeed, several of these species are sufficiently closely related that they can be hybridized. By analysing the characteristics of the progeny of such crosses (e.g. with respect to fruit composition), it is possible to make deductions about which genes may be contributing to particular traits. Using this approach, we will investigate whether any of the genes correlated with differences in fruit composition encode likely tonoplast membrane proteins. If they do, we will cross-reference this list against the information on tonoplast proteins obtained by mass spectrometry. This will allow us to focus on a limited number of the most promising candidates for more detailed characterization. In the final part of the project, we will test the function of the selected candidate proteins directly to determine, first, whether they indeed reside in the tonoplast membrane in intact cells, and second, what solutes they are capable of transporting into and out of the vacuole. We will focus on candidate transporters of organic acids and amino acids, as these are important determinants of fruit flavour and acidity that have been little investigated to date. The combination of the protein identification and genetic approaches promises to yield important new information on the factors determining fruit composition. This will also be valuable for directing future breeding strategies towards the selection of new elite lines with improved fruit traits, without the need for intervention using genetic modification techniques.

Research on particle shape is extremely important to many industrial applications such as pharmaceuticals, biopharmaceuticals, human health products and speciality chemicals. For example, for pharmaceuticals, the morphology can affect important properties such as dry powder density, cohesion, and flowability, that can have major impact on a company's ability to formulate drug particles into finished products. Moreover, crystal morphology can affect drug dissolution, potentially affecting formulated product bioavailability and, in extreme, resulting in a companies loss of the license to making the drug product. However, despite the availability of various Process Analytical Technology (PAT) instruments for measuring other properties of particulate systems, there have been no effective on-line instruments capable of providing real-time information on particle shape during the processing of particles in unit operations such as crystallisation, precipitation, granulation and milling. In the past few years, on-line high speed imaging has shown to be a very promising PAT instrument for real-time measurement of particle shape on-line which has resulted in the development of some new instrumentation products just released to the market such as the PVM (Process Vision system) of Lasentec (uk.mt.com), the PIA (Process Image Analyser) of MessTechnik Schwartz GmbH (www.mts-duesseldorf.de), the ISPV (In-Situ Particle Viewer) of Perdix (www.perdix.nl) in Netherlands, and the On-line Microscopy systems of GlaxoSmithKline, some of which incorporate a probe design which allows easy access to a processing reactor vessel. However, all these techniques are essentially limited in that they can only provide 2D information of the particle shape. Hence, this proposed research aims to develop a new instrument Stereo Vision Probe which can directly image the full 3D shape of particles within a practical processing reactor. This basic mode of operation is based on the mathematical principle that if the 2D images of an object are obtained from two different angles, the full 3D particle shape can be recovered. The potential impact on research capability and industrial applications is predicted to be major but the proposed research will focus on the development of the Stereo Vision Probe and the 3D construction method from the two 2D images obtained from two different angles. The testing of the system will be mainly via the use of a variable temperature crystallisation cell.

Doctoral Training Partnerships: a range of postgraduate training is funded by the Research Councils. For information on current funding routes, see the common terminology at www.rcuk.ac.uk/StudentshipTerminology. Training grants may be to one organisation or to a consortia of research organisations. This portal will show the lead organisation only.

An undeniable and expensive problem of any agricultural practice is the adaptation of weeds to these agricultural systems. This problem is further amplified by climate change, weeds adapt very quickly to environmental change including heat stress. The sustainable intensification of food production necessary to feed the world's growing population will only be achievable if crop harvest losses can be minimised. About 10% of crop production is currently lost to weeds and this loss would be far greater without the use of herbicides. However, the continued effectiveness of herbicide technology is threatened by the rapid advance of weed biotypes that are resistant to herbicides. Resistant weeds are now a problem across most chemical classes of herbicide and also across all the major cropping regions. Thus weed control represents a major concern for global food security, especially given that no new herbicide modes of action have been commercialized since the 1980s. New weed control tools are urgently required especially in the UK where EU/UK regulations have caused shrinkage in our 'crop protection toolbox' ("Healthy Harvest" initiative, UK National Farmers Union). Investment in crop protection in Europe has fallen from 33.3% of worldwide investment in the 1980s to 7.7% today. There is an urgent need for the development of new active substances which are effective and environmentally safe for the production of healthy food. This is also important for food quality and for preventing further food price rises which have recently affected UK consumers more than those in the mainland Europe. The problem of effective weed control is most severe in annual field crop systems and with annual weeds which emerge at the same time as the crop seedlings. These problem weeds owe their success, at least in part, to the formation of large and persistent soil seed banks. The premise of the current application is that novel and effective weed control tools might be found in compounds that either 1) promote the coordinated germination of weed seeds in the soil bank and/or 2) in 'seedicides' that kill weed seeds at the ungerminated or a very early germination stage. In the former case, germination-promoting compounds might be applied prior to conventional herbicides (or alternative non-chemical strategies) to achieve more effective control of the total weed seed population while 'seedicides' might be applied after crops have emerged in order to limit late weed emergence. Currently, while some germination stimulants are known, none are commercial and neither do any commercial herbicides deliberately target seed-specific processes. All classical herbicides target processes of growing weed seedlings. Thus there is considerable potential for novel weed control solutions through engaging a deeper understanding of the processes of weed seed germination, survival and persistence. This project establishes a collaboration between the Seed Biology Group of Prof G Leubner at Royal Holloway University of London and Weed Control Research Biology at Syngenta's International Research Centre Jealott's Hill. We plan to investigate the described issues with several representative problem weeds by first modelling their germination responses to a wide range in ambient temperatures, and then by choosing key temperatures to investigate the underlying hormonal changes and molecular mechanisms without and with the application of compounds known to break dormancy and induce germination of seeds. The obtained knowledge will be used to select compound libraries and screen for novel chemicals which have the potential to either promote the coordinated germination of weed seeds and/or to act as 'seedicides'. This will potentially identify novel modes of action instrumental for downstream research to fill our 'crop protection toolbox' with novel chemicals. Our project will generate knowledge to guide the development of novel strategies to control weeds by depleting the seed bank.

Doctoral Training Partnerships: a range of postgraduate training is funded by the Research Councils. For information on current funding routes, see the common terminology at www.rcuk.ac.uk/StudentshipTerminology. Training grants may be to one organisation or to a consortia of research organisations. This portal will show the lead organisation only.