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Optimisation of high-value industrial and nutritional carotenoids in plant based renewable sources.


Project Description

Background and rationale
Carotenoids are high-value antioxidant compounds used across the food, feed, cosmetic, health and pharma sectors. They are also essential components of the human diet. Tomato fruit and its products are the major source of bioactive carotenoids in the diet and more recently tomato has been used as a renewable source for these compounds. The applicants have shown, by genetic intervention (transgenesis, TILLING and marker assisted selection), that the carotenoid content of tomato fruit can be increased to: (i), improve nutritional and aesthetic quality and (ii), provide new, renewable sources of high-value industrial carotenoids. In the latter case, in addition to the improved environmental credentials, the levels can compete economically with petrochemical derived synthesis.
Despite these advances in increasing or changing the carotenoids in fruit, biochemical characterisation of the new varieties has revealed a significant role for carotenoid degradation (e.g. the release of volatile and semi-volatile derived products) in modulating carotenoid accumulation. The extent to which catabolism regulates the carotenoid content in fruit awaits elucidation. This important aspect of regulation requires further research and will be the focus of the proposed project, as we attempt to further optimise nutritional and industrial carotenoid contents in crop plants by deducing the role of carotenoid degradation in the accumulation and maintenance of carotenoid homeostasis.
Aims and objectives
Aim: The HIGHCAR project will elucidate the role of the catabolism in modulating carotenoid content and apocarotenoid production in ripe tomato fruit. To achieve this goal, the following objectives will be performed; (i) quantitative determination of apocarotenoids and transcripts for Carotenoid Cleavage Oxygenases (CCO) present in varieties with with altered carotenoid content, (ii) generate transgenic tomato plants where carotenoid metabolising enzymes have been down-regulated, (iii) perform molecular and biochemical characterisation of the transgenic genotypes produced
Experimental approach. The group has developed a unique genetic resource of stable tomato genotypes with altered carotenoid contents. These varieties will be used to ascertain the relationship between isoprenoids, carotenoids and their derived apocarotenoid catabolic metabolites. Metabolite profiling using LC-MS/MS, GC-MS and HPLC/ULC-PDA platforms will be used to ascertain apocarotenoid contents. The platforms are established in the laboratory with experienced personnel capable of providing in-house training and access to customised metabolite libraries. QRTPCR and proteomics will be used to determine the changes in transcripts and proteins that resulted from altered carotenoid contents. The applicants laboratory has adopted Synthetic Biology high-throughput cloning approaches. The experimental approach proposed will provide the prospective student with the opportunity to develop generic skillsets in biochemistry (metabolite profiling) and advanced molecular (Synthetic ) biology. The outputs have direct industrial applications.
Environment. The applicants’ laboratory is a well-equipped and funded environment participating in COST ACTIONS, and three integrated EU networks, two coordinated by the applicants. These networks provide excellent training opportunities and access to specialist advice and facilities. The group has dedicated analytical apparatus; GC-MS (x 3), GC-FID, HPLC-PDA (x2), a HPLC-PDA-radiodetector, UPLC-PDA and real-time PCR machines. Growth facilitates include a glasshouse (600m2), tissue culture facilities and a polytunnel (300m2). The proposed project will be embed into the Horizon2020 funded TomGem project and BBSRC-OptiCAR projects. The student will work with the laboratory teams actively engaged in these funded projects.

Funding Notes

Applicants should already have or be expected to obtain a First or upper Second Class degree in a relevant discipline. This studentship is fully funded for three years. It covers tuition fees at the UK/EU rate and includes a stipend at the standard Research Council rate (currently £16,296 per annum). Funding is available for UK and EU students.

References

1. Fraser P.D., Enfissi .E.M.A., Halket J.M, Truesdale, M.R., Yu. D., Gerrish, C. and Bramley P.M. (2007). Manipulation of phytoene levels in tomato fruit: effects on isoprenoids, plastids and intermediary metabolism. Plant Cell, 19: 3194-3211.

2. Enfissi, E.M.A., Barneche, F., Ahmed, I., Lichtlé, C., Gerrish, C., McQuinn, R.P., Giovannoni, J.J., Lopez-Juez, E., Bowler, C., Bramley, P.M., and Fraser, P.D.** (2010). Integrative transcript and metabolite analysis of DE-ETIOLATED1 down regulated tomato fruit reveals the underlying metabolic and cellular events associated with their nutritionally enhanced chemotype. Plant Cell, 22, 1190-1215.

3. Nogueira, M., Mora, L., Enfissi, E.M.A., Bramley, P.M. and Fraser,P.D.** (2013). Subchromoplast Sequestration of Carotenoids Affects Regulatory Mechanisms in Tomato Lines Expressing Different Carotenoid Gene Combinations. Plant Cell, 25, 4560-4579.

4. Perez-Fons, L., Wells, T., Coral, D.I., Ward, J.L., Gerrish, C., Beale, M.H., Seymour, G.B., Bramley, P.M. and Fraser, P.D.**(2014). A genome-wide metabolomics resource for tomato fruit from Solanum pennellii. Scientific Reports 4, Article number 3859.

5. Uluisik, S., Chapman, M.H., Smith, R., Poole, M., Adams, G., Gillis, R.B., Besong, T.M.D., Sheldon,. J., Stiegemeyer, S., Perez-Fons, L., Samsulrizal, N., Wang, D., Fisk, I.D. Yang, Ni, Baxter, C., Rickett, D., Fray, R., Blanco-Ulate, B., Powell, A.L., Harding, S.E., Craigon, J., Rose, K.C. J., Fich, E.A., Sun, Li, Fraser, P.D., Tucker, G.A., Grierson, D., and Seymour, G.B. (2016). Genetic improvement of tomato by targeted control of fruit softening Nature Biotechnology. 34, 950–952.

6. Price, E.J., Wilkin, P., Sarasan, W., and Fraser, P.D. (2016). Metabolite profiling of Discorea (yam) species reveals underutilised biodiversity and renewable source for high-value compounds. Scientific Reports, 6: 29136.

7. Enfissi, E.M.A., Nogueira, M., Bramley, P.M. and Fraser, P.D. (2016). New insights into the regulation of carotenoid biosynthesis in tomato fruit. Plant J. In press.

How good is research at Royal Holloway, University of London in Biological Sciences?

FTE Category A staff submitted: 24.00

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