| Experiment: | 29 |
| Title: | Identification of genes controlling stomatal number in response to rising atmospheric carbon dioxide |
| Date: | 2002-11-08 |
| Description: | As part of a GUS promoter trap screen a gene exhibiting a guard cell-specific expression pattern was identified. When these plants are grown at elevated (1000ppm) CO2 they display a massive increase in stomatal density compared with the parent ecotype (C24) a response markedly different to the majority of plant species which show a decrease in stomatal density in response to elevated CO2. The disrupted gene has been identified and named HIC (for High CO2; Gray et al.2000 Nature 408: 713-716). Using the GARNet transcriptome analysis facility we shall be able to rapidly identify genes whose expression is altered between guard cells of hic and C24 (parental ecotype) plants in response to elevated CO2 levels. Control plants will be grown under identical light (240 mM m-2 s-1; 18hr light/6hr dark) and temperature (22oC light; 18oC dark) regimes but at ambient (360ppm) CO2 and guard cell enriched extracts produced from developing leaves. We will then be able to compare the expression profiles of C24 and hic guard cells grown under ambient and elevated atmospheric CO2 to identify additional components of this environmental signalling pathway. This approach should identify both genes involved in responses to rising atmospheric CO2 levels and genes involved in the control of stomatal development. This is a BBSRC collaborative project with Prof. A. Hetherington's laboratory, Institute of Environmental and Natural Sciences, University of Lancaster and Prof. F. I. Woodward, APS, University of Sheffield. Plants will be grown at Lancaster in controlled environment growth chambers (Percival Scientific, IA, USA) providing 240 mM m-2 s-1 18hr light/6hr dark regime at 22oC (light) 18oC (dark) with RH at 60 % +. Seeds will be surface sterilised using 10 % domestic bleach (<0.5 % sodium hyochlorite solution), washed with sterile RO water and sowed onto half-strength (2.2 g/l) Murashigee and Skoog basal media (Sigma chemical M5519), 10 g/l (1 %) sucrose, solidified with 6 g/l (0.6 %) plant cell tissue grade agar (Sigma chemical A1296). Media will be contained in 10 cm 3-vent petri dishes that are sealed with microporous surgical tape after sowing. All sown plates will be given a 48-hour stratification treatment at 0-4oC before going into growth chambers.After 10 days, the seedlings will be transferred to a peat based compost mixture (4 parts SHL multipurpose peat based compost (William Sinclair Horticulture, Lincoln, UK): 1 pat washed horticultural silver sand (GEM Horticulture, Accrington, Lancs., UK). After a further seven days plants are repotted (4:1 SHL: GEM silver sand) into 5 cm Arabaskets, of the Arasystem, Arabidopsis growing kit (Beta Tech, Gent, Belgium). At 5 weeks old, before inflorescence bolting had occurred, all leaf material from 10 plants (whole leaf) and 20 plants (guard cell enriched) will be harvested and cryogenically stored. RNA will be extracted in Sheffield using TRIzol reagent (GIBCo BRL) and cleaned using Qiagen minicolumns. |
| ftp Link: | ftp Link |
| Slide ID | Slide Name | Genetic Background | Tissue | Stock Code | Cel File |
|---|
| Heggie_1-10_C24-ambientCO2-leaf-(CWF)_Rep3_ATH1 | 782 | | whole leaves | N906 | LH001_ATH1_A2-HEGGI-CWF.CEL |
| Heggie_1-11_C24-ambientCO2-guard-cell-(CGC)_Rep2_ATH1 | 783 | | Guard Cell enriched leaves | N906 | LH001_ATH1_A3-HEGGI-CGC.CEL |
| Heggie_1-12_C24-ambientCO2-guard-cell-(CGL)_Rep3_ATH1 | 784 | | Guard Cell enriched leaves | N906 | LH001_ATH1_A4-HEGGI-CGL.CEL |
| Heggie_1-1_C24-ambientCO2-guard-cell-(CAG)_Rep1_ATH1 | 206 | | Guard Cell enriched leaves | N906 | LH001_ATH1_A1-Heggi-CAG.CEL |
| Heggie_1-2_C24-elevatedCO2-guardcell_(CEG)_Rep1_ATH1 | 208 | | Guard Cell enriched leaves | N906 | LH001_ATH1_A2-Heggi-CEG.CEL |
| Treatment: This sample was grown at elevated CO2 (1000ppm) |
| Heggie_1-3_HIC-ambientCO2-guard-cell-(HAG)_Rep1_ATH1 | 210 | C24, N906 | Guard Cell enriched leaves | | LH001_ATH1_A3-Heggi-HAG.CEL |
| Heggie_1-4_HIC-elevatedCO2-guard-cell-(HEG)_Rep1_ATH1 | 212 | C24, N906 | Guard Cell enriched leaves | | LH001_ATH1_A4-Heggi-HEG.CEL |
| Treatment: This sample was grown at elevated CO2 (1000ppm) |
| Heggie_1-5_C24-ambientCO2-leaf-(CAW)_Rep1_ATH1 | 207 | | whole leaves | N906 | LH001_ATH1_A5-Heggi-CAW.CEL |
| Heggie_1-6_C24-elevatedCO2-leaf-(CEW)_Rep1_ATH1 | 209 | | whole leaves | N906 | LH001_ATH1_A6-Heggi-CEW.CEL |
| Treatment: This sample was grown at elevated CO2 (1000ppm) |
| Heggie_1-7_HIC-ambientCO2-leaf-(HAW)_Rep1_ATH1 | 211 | C24, N906 | whole leaves | | LH001_ATH1_A7-HEGGIE-HAW_repeat.CEL |
| Heggie_1-8_HIC-elevatedCO2-leaf-(HEW)_Rep1_ATH1 | 213 | C24, N906 | whole leaves | | LH001_ATH1_A8-HEGGIE-HEW_repeat.CEL |
| Treatment: This sample was grown at elevated CO2 (1000ppm) |
| Heggie_1-9_C24-ambientCO2-leaf-(CWL)_Rep2_ATH1 | 781 | | whole leaves | N906 | LH001_ATH1_A1-HEGGIE-CWL_repeat.CEL |
NASCArrays Information at The BAR
Welcome to NASCArrays information at the BAR. This page hosts meta-information from the NASCArrays service (2002-2013). This information was parsed
from text files available on the NASCArrays site. NASCArrays data is on iPlant server. To download experiment data from iPlant, please click on the
experiment number. To download the CEL files, please click on the ftp link.