FP6Individual fellowship2004–2006

GSH - GROWTH · Identifying genes involved in environmental sensing and plant growth

FP6 — Marie Curie Actions (Human Resources and Mobility)

Duration
2004-08-01 → 2006-07-31
EU contribution
€170,500
Participants
1
Scheme
IIF

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Results in brief

Final Activity Report Summary - GSH - GROWTH (Identifying genes involved in environmental sensing and plant growth)

The link between the key cellular antioxidant, glutathione (GSH), and growth of the primary root in plants was investigated using molecular, genomic and genetic techniques. A series of studies were devised and performed on cell cultures using a chemical inhibitor to deplete GSH in living cells to examine the effects of low GSH on cell division. An exciting outcome of these experiments was the demonstration that for the thale cress, Arabidopsis, it appears that GSH is required only when cells are dividing within a developmental context indicating that perhaps it is acting as a signalling component rather than as a detoxifying agent. By analysing microarray data from root tips and data provided by an industry-linked collaboration this project broke new ground on the GSH growth pathway. This revealed novel genes which may mitigate root growth in response to GSH levels, and these predictions were tested experimentally to delineate a heretofore unknown element in GSH sensing.

Data: CORDIS, © European Union

Project objective

Glutathione (GSH) is an important and abundant reducing agent in plants. GSH is involved in tolerance and adaptation toenvironmental stresses, including drought, pollution and temperature extremes.The rate-limiting step in GSH production is catalysed by ga mma-glutamylcysteine synthetase (GCS). In Arabidopsis, twoGSH-deficient mutants define a single locus, ROOT MERISTEMLESSI (RML1)/CADMIUM SENSITIVE2 (CAD2),encoding GCS. The rmll mutant contains 2% of wild-type GSH levels and although embryonic development is normal,on germination, rmll root cells fail to divide and consequently do not maintain the root meristem. In contrast, shootdevelopment proceeds relatively normally. The rmll phenotype is rescued by exogenous GSH (but not other reductants)indicating the specific importance of GSH in root cell division. Moreover, wild-type plants phenocopy rmll whendepleted of GSH by a specific inhibitor of GCS, buthionine sulfoximine (BSO). In tobacco cell culture, BSO treatmentcauses a specific arrest in cell division a t a Gl checkpoint during the cell cycle. These results show a clear link betweencell cycle progression and GSH levels in cell cultures and in the root meristem. Here we propose to analyse its nature andmechanism by (a) establishing which known cell cycle g enes respond to GSH levels, (b) identifying novel genes that areaffected by GSH levels by using microarrays and (c) analysing the novel genes to find which are upstream regulators ofcell cycle targets. To achieve these aims we will use the specific GCS inh ibitor, buthionine sulfoximine, to deplete GSHlevels in the novel synchronizable MM1 and MM2d Arabidopsis cell lines in combination with the rmll and cad2mutants and examine gene expression. During this project the candidate will gain valuable scientific t raining (ingenomics, bioinformatics and cell biology) and professional training (through presentations and the host institution'spersonal development #'

Original text from CORDIS.

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Data: CORDIS, © European Union