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Experiment: The trans-differentiation of cultured Arabidopsis cells
Experiment Description
NASCArrays Experiment Reference Number: NASCARRAYS-80
The formation of vascular tissue occurs when cellulose, hemicellulose, lignin and other wall components are deposited within the primary cell wall. These secondary thickened cells then undergo programmed cell death producing a network of empty cells with which water and ions can be transported throughout the plant. The hormones auxin and cytokinin are the principle signals for vascular tissue initiation. As a consequence cells cultured in-vitro can be converted into vascular tissue with the addition of exogenous auxin and cytokinin.
We have created an in-vitro cell system, using callus produced from leaves that can be induced to form vascular tissue. Leaves are callused on induction media for two weeks. The callus is then transferred to liquid media and incubated under optimum conditions resulting in an increase in vascular tissue formation. Approximately 20% of cells will differentiate during the incubation period. The alteration of cytokinin concentration affects the ability of the cultured cells to undergo differentiation. Consequently callus incubated in liquid media, containing lower cytokinin concentrations, will undertake relatively little differentiation. Samples have been isolated from cell cultures at different time points and different hormone concentrations during incubation. Quantitative PCR using the marker AtCesA7, which encodes a cellulose synthase subunit specific to secondary wall deposition, was used as a guide to determine periods of high and low vascular differentiation. This system provides an opportunity to compare gene expression between differentiating and non differentiating cells and allow the identification of genes up regulated during vascular tissue formation.
About the ExperimenterName: | Mr David Brown |
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Head of Lab Name: | Dr Simon Turner |
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Lab:
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Institute:
| University of Manchester |
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Address: | University of Manchester School of biological Sciences 3.614 Stopford Building Oxford Road Manchester
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Postcode:
| M13 3PT |
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Country:
| UK |
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| Telephone Number:
| 0161 2755109 |
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Fax Number:
| 0161 2753938 |
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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:
| time_series_design; compound_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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parameter | timepoint |
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Quality Control Measures Taken:
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References:
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| Other Information:
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ArrayExpressAccession | E-NASC-15 |
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Slides in this Experiment
Hybridisation Set: Brown_genome
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Tissue:
| Cell culture |
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Diseased:
| Normal |
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in vitro Treatment:
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treatment | Leaves from seedlings, grown aseptically (14-17 days) on MS agar, excised and transferred to callus induction medium (Gamborg's B5 Basil Medium with minimal organics, 2.0% glucose, 0.5mgl-1 MES, 0.5mgl-1 2,4-D, 0.05mgl-1 Kinetin. pH 5.7). Incubated in darkness, at 22oC for 2 weeks. Callus not transferred to liquid media. RNA collected therefore t=0. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: A-2-Brown-2day_high | | |
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Tissue:
| Cell culture |
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Diseased:
| Normal |
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in vitro Treatment:
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treatment | Leaves from seedlings, grown aseptically (14-17 days) on MS agar, excised and transferred to callus induction medium (Gamborg's B5 Basil Medium with minimal organics, 2.0% glucose, 0.5mgl-1 MES, 0.5mgl-1 2,4-D, 0.05mgl-1 Kinetin. pH 5.7). Incubated in darkness, at 22oC for 2 weeks. Callus transferred to high differentiating liquid media (MS 0.2% sucrose, 4% mannitol, 0.2mgl-1 auxins (2,4-D, NAA, IAA) 1mgl-1 BA, pH5.7). Incubated at 22oC on a rotary shaker (105rpm). Sample taken at day =2 |
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Other Information:
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Protocols for BioSource 1 |
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Slide: A-3-Brown-5day_high | | |
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Tissue:
| Cell culture |
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Diseased:
| Normal |
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in vitro Treatment:
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treatment | Leaves from seedlings, grown aseptically (14-17 days) on MS agar, excised and transferred to callus induction medium (Gamborg's B5 Basil Medium with minimal organics, 2.0% glucose, 0.5mgl-1 MES, 0.5mgl-1 2,4-D, 0.05mgl-1 Kinetin. pH 5.7). Incubated in darkness, at 22oC for 2 weeks. Callus transferred to high differentiating liquid media (MS 0.2% sucrose, 4% mannitol, 0.2mgl-1 auxins (2,4-D, NAA, IAA) 1mgl-1 BA, pH5.7). Incubated at 22oC on a rotary shaker (105rpm). Sample taken at day =5 |
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Other Information:
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Protocols for BioSource 1 |
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Slide: A-4-Brown-9day_high | | |
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Tissue:
| Cell culture |
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Diseased:
| Normal |
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in vitro Treatment:
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treatment | Leaves from seedlings, grown aseptically (14-17 days)on MS agar, excised and transferred to callus induction medium (Gamborg's B5 Basil Medium with minimal organics, 2.0% glucose, 0.5mgl-1 MES, 0.5mgl-1 2,4-D, 0.05mgl-1 Kinetin. pH 5.7). Incubated in darkness, at 22oC for 2 weeks. Callus transferred to high differentiating liquid media (MS 0.2% sucrose, 4% mannitol, 0.2mgl-1 auxins (2,4-D, NAA, IAA) 1mgl-1 BA, pH5.7). Incubated at 22oC on a rotary shaker (105rpm). Sample taken at day =9 |
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Other Information:
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Protocols for BioSource 1 |
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Slide: A-5-Brown-5day_low | | |
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Tissue:
| Cell culture |
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Diseased:
| Normal |
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in vitro Treatment:
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treatment | Leaves from seedlings, grown aseptically (14-17 days)on MS agar, excised and transferred to callus induction medium (Gamborg's B5 Basil Medium with minimal organics, 2.0% glucose, 0.5mgl-1 MES, 0.5mgl-1 2,4-D, 0.05mgl-1 Kinetin. pH 5.7). Incubated in darkness, at 22oC for 2 weeks. Callus transferred to low differentiating liquid media (MS 0.2% sucrose, 4% mannitol, 0.5mgl-1 2,4-D, 1.5ugl-1 Kinetin, pH5.7). Incubated at 22oC on a rotary shaker (105rpm). Sample taken at day =5 |
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Other Information:
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Protocols for BioSource 1 |
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Slide: A-6-Brown-9day_low | | |
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Tissue:
| Cell culture |
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Diseased:
| Normal |
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in vitro Treatment:
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treatment | Leaves from seedlings, grown aseptically (14-17 days)on MS agar, excised and transferred to callus induction medium (Gamborg's B5 Basil Medium with minimal organics, 2.0% glucose, 0.5mgl-1 MES, 0.5mgl-1 2,4-D, 0.05mgl-1 Kinetin. pH 5.7). Incubated in darkness, at 22oC for 2 weeks. Callus transferred to low differentiating liquid media (MS 0.2% sucrose, 4% mannitol, 0.5mgl-1 2,4-D, 1.5ugl-1 Kinetin, pH5.7). Incubated at 22oC on a rotary shaker (105rpm). Sample taken at day =9 |
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Other Information:
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Protocols for BioSource 1 |
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