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Experiment: Identification of genes responsive to non-metabolised glucose analoges as an approach to hexokinase-independent glucose sensing in plants.
Experiment Description
NASCArrays Experiment Reference Number: NASCARRAYS-40
It has been strongly argued that plant cells should have a means of sensing sugars at the cell surface, so that extracellular and intracellular sugars can be sensed separately and their metabolism coordinated (Lalonde et al., Plant Cell, 11, 707-26, 2000). There is good evidence for an intracellular hexokinase-dependent pathway of hexose sensing in plants, but very little evidence for a hexokinase-independent signalling pathway, such as that provided by SNF3 or RGT2 in yeast. Many papers on sugar sensing in plants cite work from two laboratories as evidence for hexokinase-independent hexose signalling in plants. The first is that in which cell-wall invertase and sucrose synthase genes were induced by treatment of a Chenopodium suspension culture with 30 mM 6-Deoxyglucose (6DOG) for 24 h (Roitsch et al., Plant Physiol 108, 285-294, 1995; Godt et al., J. Plant Physiol 146, 231-238, 1995). The second is that in which a patatin transgene in Arabidopsis was shown to be weakly induced by growth over several days on a mixture of 30 mM glucose plus 30 mM 3-O-methylglucose (3OMG), but strongly induced by growth on 30 mM Glc plus 90 mM 3OMG (Martin et al., Plant J, 11, 53-62, 1997). We are not aware of any examples of Arabidopsis genes which respond to 6DOG or 3OMG yet this is an area of wide significance. Identification of such a gene would help to establish if a hexokinase-independent signalling system operates in plants, and would provide a basis for establishment of a genetic screen for mutants, using the gene promoter linked to a reporter such as luciferase. The aim of this proposal is to discover any genes which are either activated or repressed by glucose AND by 3OMG and/or 6DOG, but not by mannitol (an osmotic control). The use of both 3OMG and 6DOG will help to identify non-specific effects of either. All substrates will first be analysed by HPLC to confirm that they are pure. Arabidopsis Col-0 seedlings will be grown in vitro for 7 days in the absence of sugars, then treated with 30 mM glucose or glucose analogue for 8 h (these conditions are based on concentrations and time courses of Roitsch et al.). RNA will then be isolated from multiple independent plates to minimise biological variation.
About the ExperimenterName: | Dr Dorthe Villadsen |
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Head of Lab Name: | Dr Steve Smith |
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Lab:
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Institute:
| University of Edinburgh |
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Address: | Institute of Cell and Molecular Biology University of Edinburgh The King's Buildings Mayfield Road Edinburgh
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Postcode:
| EH9 3JH |
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Country:
| UK |
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| Telephone Number:
| 0131 650 5318 |
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Fax Number:
| 0131 650 5392 |
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All of the data available in this website/database is free, and you
are free to do whatever you please with it. If you intend to publish
work based on any of this data, please acknowledge us, contact the
experimenter above, and either acknowledge them or use them as
co-authors in the work.
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| About this ExperimentExperiment Type:
| compound_based_treatment_design |
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Number of Slides: | 6 |
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| Experimental Parameters:
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Parameter | compound_based_treatment |
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Quality Control Measures Taken:
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no-plants-pooled | 100s |
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other | Three separate plates pooled for each sample |
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References:
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| Other Information:
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ArrayExpress Accession | E-NASC-8 |
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Slides in this Experiment
Hybridisation Set: Villadsen_genome
Slide: Villadsen_A-1_zer_Rep1_ATH1 | | |
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Tissue:
| the entire seedlings |
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Plant Ontology terms for this tissue type: | |
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| | | Unknown PO Term! | Diseased:
| Normal |
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in vitro Treatment:
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treatment | The seedlings were incubated in horizontal position for 8 hours under 150 umol/m2/s, no other treatment |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Villadsen_A-2_wat_Rep1_ATH1 | | |
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Tissue:
| the entire seedlings |
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Plant Ontology terms for this tissue type: | |
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| | | Unknown PO Term! | Diseased:
| Normal |
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in vitro Treatment:
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treatment | The seedlings were incubated in horizontal position for 8 hours under 150 umol/m2/s, with 15 mL water.
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Other Information:
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Protocols for BioSource 1 |
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Slide: Villadsen_A-3_glc_Rep1_ATH1 | | |
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Tissue:
| the entire seedlings |
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Plant Ontology terms for this tissue type: | |
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| | | Unknown PO Term! | Diseased:
| Normal |
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in vitro Treatment:
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treatment | The seedlings were incubated in horizontal position for 8 hours under 150 umol/m2/s, with 15 mL 30 mM glucose |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Villadsen_A-4_OMG_Rep1_ATH1 | | |
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Tissue:
| the entire seedlings |
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Plant Ontology terms for this tissue type: | |
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| | | Unknown PO Term! | Diseased:
| Normal |
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in vitro Treatment:
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treatment | The seedlings were incubated in horizontal position for 8 hours under 150 umol/m2/s, with 15 mL 30 mM 3-O-methylglucose |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Villadsen_A-5_DOG_Rep1_ATH1 | | |
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Tissue:
| the entire seedlings |
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Plant Ontology terms for this tissue type: | |
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| | | Unknown PO Term! | Diseased:
| Normal |
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in vitro Treatment:
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treatment | The seedlings were incubated in horizontal position for 8 hours under 150 umol/m2/s, with 15 mL 30 mM 6-deoxyglucose |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Villadsen_A-6_man_Rep1_ATH1 | | |
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Tissue:
| the entire seedlings |
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Plant Ontology terms for this tissue type: | |
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| | | Unknown PO Term! | Diseased:
| Normal |
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in vitro Treatment:
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treatment | The seedlings were incubated in horizontal position for 8 hours under 150 umol/m2/s, with 15 mL 30 mM manitol |
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Other Information:
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Protocols for BioSource 1 |
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