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Transcriptome analysis of pho3

ABSTRACT: The aim of this proposal is to understand the impact of the pho3 mutation on the transcriptome.The mutant pho3 was isolated on the basis of a failure to induce acid phosphatase activity in roots in response to low Pi. The pho3 mutant had a P deficient phenotype in vitro and in the leaves of soil grown pho3 plants the total P content was again reduced. pho3 exhibited a number of characteristics normally associated with low-P stress, including severely reduced growth, increased anthocyanin content and starch accumulation. pho3 was shown to have an absolute requirement for exogenous sucrose as pho3 seedlings were arrested in early seedling development on sucrose-free medium and was unaffected by Pi-availability. pho3 was found to be insensitive to high concentrations of exogenous sucrose (10%) but was sensitive to glucose. pho3 also exhibited reduced sensitivity of seed germination to exogenous ABA (abi-phenotype), a response which has been linked to sucrose insensitivity in both sugar-sensing and ABA mutants. Two developmental stages have been selected on the basis of the physiological characterisation of pho3. Seedlings (1.02; 2 rosette leaves) grown in agar and compost-grown plants (3.90; rosette growth complete). Agar will be supplemented with 1% sucrose and 1/2 strength MS; plants will be grown under a 16 h light/8 h dark cycle. Comparisons will be made between pho3 and Wassilewskija (the genetic background of the mutant) at each stage. Experimenter name: Julie Lloyd Experimenter phone: 01206 873307 Experimenter institute: Department of Biological Sciences, University of Essex Experimenter address: Colchester!Series_summary = Experimenter zip/postal_code: CO4 3SQ Experimenter country: UK Keywords: genetic_modification_design

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE10326 | GEO | 2008/01/31

SECONDARY ACCESSION(S): PRJNA108565

REPOSITORIES: GEO

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Project description:The aim of this study is to study gene expression in Brassica oleracea in shoot tissues of plants grown under contrasting P supplies (see Hammond JP et al., 2003, Plant Physiology, 132, 578-596 for background). Seeds of B. oleracea (var. alboglabra, A12dH) were first washed in 70% (v/v) ethanol/water, rinsed in distilled water and surface sterilised using 50% (v/v) domestic bleach/water. Seeds were rinsed and imbibed for 3 to 5 days in sterile distilled water at 4°C to break dormancy. Following imbibition, B. oleracea seeds were sown in un-vented, polycarbonate culture boxes (Sigma-Aldrich Company Ltd., Dorset UK). Seedlings were grown for 21 days on perforated polycarbonate discs (diameter 91 mm by 5 mm) placed on 75 ml of 0.8% (w/v) agar containing 1% (w/v) sucrose and a basal salt mix. Roots grew into the agar, but shoots remained on the opposite side of the disc. After 21 days, seedlings were transferred, still on polycarbonate discs, to a hydroponics system situated in a Saxcil growth cabinet (16 h daylength, set to 22°C and 80% humidity). Each polycarbonate disc was placed on a light-proof 500 ml beaker over 450 ml of nutrient solution. After 7 days, half the plants were transferred to nutrient solution containing no phosphate and the other half remained on full nutrient solution (control plants). Shoots were harvested 100 h after the withdrawal of P. !Samples were snap frozen in liquid nitrogen. RNA was extracted using the TRIzol extraction method and cleaned through a Qiagen RNeasy column. Experimenter name = Martin Broadley Experimenter phone = 0115 951 6382 Experimenter fax = 0115 951 6334 Experimenter institute = University of Nottingham Experimenter address = Plant Sciences Division Experimenter address = School of Biosciences Experimenter address = University of Nottingham Experimenter address = Sutton Bonington Experimenter address = Loughborough Experimenter zip/postal_code = LE12 5RD Experimenter country = UK Keywords: growth_condition_design

Project description:The aim of this study is to study gene expression in Brassica oleracea in shoot tissues of plants grown under contrasting P supplies (see Hammond JP et al., 2003, Plant Physiology, 132, 578-596 for background). Seeds of B. oleracea (var. alboglabra, A12dH) were first washed in 70% (v/v) ethanol/water, rinsed in distilled water and surface sterilised using 50% (v/v) domestic bleach/water. Seeds were rinsed and imbibed for 3 to 5 days in sterile distilled water at 4Â°C to break dormancy. Following imbibition, B. oleracea seeds were sown in un-vented, polycarbonate culture boxes (Sigma-Aldrich Company Ltd., Dorset UK). Seedlings were grown for 21 days on perforated polycarbonate discs (diameter 91 mm by 5 mm) placed on 75 ml of 0.8% (w/v) agar containing 1% (w/v) sucrose and a basal salt mix. Roots grew into the agar, but shoots remained on the opposite side of the disc. After 21 days, seedlings were transferred, still on polycarbonate discs, to a hydroponics system situated in a Saxcil growth cabinet (16 h daylength, set to 22Â°C and 80% humidity). Each polycarbonate disc was placed on a light-proof 500 ml beaker over 450 ml of nutrient solution. After 7 days, half the plants were transferred to nutrient solution containing no phosphate and the other half remained on full nutrient solution (control plants). Shoots were harvested 100 h after the withdrawal of P. !Samples were snap frozen in liquid nitrogen. RNA was extracted using the TRIzol extraction method and cleaned through a Qiagen RNeasy column. Experimenter name = Martin Broadley; Experimenter phone = 0115 951 6382; Experimenter fax = 0115 951 6334; Experimenter institute = University of Nottingham; Experimenter address = Plant Sciences Division; Experimenter address = School of Biosciences; Experimenter address = University of Nottingham; Experimenter address = Sutton Bonington; Experimenter address = Loughborough; Experimenter zip/postal_code = LE12 5RD; Experimenter country = UK Experiment Overall Design: 6 samples were used in this experiment

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Transcriptome analysis of pho3

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