Award details

Metabolic detoxification of herbicides and other xenobiotics by plants; the role of transport

Reference	BBS/E/C/00011527
Principal Investigator / Supervisor	Professor David Burt
Co-Investigators / Co-Supervisors
Institution	Rothamsted Research
Department	Rothamsted Research Department
Funding type	Research
Value (£)	269,766
Status	Completed
Type	Institute Project
Start date	01/04/1997
End date	31/03/1999
Duration	24 months

Abstract

Overall, the project will identify and isolate key proteins involved in the formation and transport of glutathione- xenobiotic conjugates. This will lead to isolation of genes for these proteins, and an understanding of factors which regulate the expression of the pathway. Initially, cereals and the weed blackgrass (A. myosuroides) will be studied, since these plants (or biotypes herbicides and other xenobiotics by conjugation to glutathione and the exclusion of these conjugates from the cytoplasm, by transport to the vacuole, will be examined. The overall pathway will be delineated at the cellular level in cultured cells and protoplasts using fluorescent and radioactive tracers. The transport mechanisms at the tonoplast will be studied in isolated vacuoles with radiolabelled substrates. Attempts will be made to identify and isolate the proteins responsible for conjugation and transport and to clone the genes involved. The biochemical and physiological information generated by this project will provide a foundation for other work on the molecular biology of plant transport systems being conducted within Programme 215 (projects 011569 and 011531). To use isolated protoplasts and vacuoles to determine the capacities of herbicide susceptible and herbicide-resistant black-grass biotypes, for glutathione conjugate formation and compartmentation. To purify and prepare homoglutathione and hydroxy-methylglutathione conjugates and to test these as substrates for the conjugate transporter. To develop a transport assay that allows the purification and identification of the proteins involved in ATP- dependent conjugate transport. To determine the range of constitutive glutathione-S-transferase activities in barley and black-grass seedlings using a range of herbicides and non- herbicide substrates. To carry out an expression analysis of the glutathione conjugate transporter genes (MRP) in barley. To optimise methods for isolation of mesophyll vacuoles from wheat leaves and to measure transport in these vacuoles with a range of herbicide glutathione conjugates. To develop a fluorescent method based on monochlorobimane to assay the glutathione mediated metabolism of herbicides, and to use these procedures to study metabolism-based herbicide resistance in blackgrass and wild oats.

Summary

unavailable

Committee	Closed Committee - Agri-food (AF)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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