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Experiment: Analysis of anther development by identifiying downstream genes controlled by MS35/MYB26
Experiment Description
NASCArrays Experiment Reference Number: NASCARRAYS-442
MALE STERILITY 35 (MS35/ MYB26) is a R2R3-type MYB transcription factor (Steiner-Large et al., 2003). MS35 is suggested to be a key regulator in the process of lignified secondary thickening in the anther endothecium which provides mechanical force for anther dehiscence (Dawson et al., 1999). The ms35 mutant bears non-dehiscent anthers and thus exhibits a male sterile phenotype. This microarray experiment aims to identify genes controlled by MS35 and thus establish the precise role MS35 plays during anther development.We have demonstrated that MS35 is expressed in the anther during pollen mitotic divisions. Comparisons of gene expression will be carried out between anthers from the Ler.gl (wild-type control) and ms35.gl (mutant), to identify genes with altered expression due to the ms35 mutation. The use of the gl marker will enable a prompt identification of the ms35 mutant amongst the segregating population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation.Plants were grown in growth chambers at 22h of daylight (167 umol.m2.sec-1) at 24 C and 2h of night at 24 C. Developmental stages of anthers were defined by bud position and DAPI staining. As soon as the plants showed three fertile or sterile siliques, anther and filaments were collected from both lines at Pollen Mitosis I (PM I), bicellular, and PM II stage, separately. RNA was extracted using RNeasy Mini Kit (Qiagen, USA) and digested with DNase to eliminate genomic DNA contamination. The identified genes will be further analysed using bioinformatics and genomic (knockout) resources that are available for Arabidopsis.
About the ExperimenterName: | Miss Jie Song |
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Head of Lab Name: | Dr Zoe Wilson |
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Lab:
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Address: | Plant Sciences University of Nottingham Sutton Bonington Campus Loughborough Leics
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Postcode:
| LE12 5RD |
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Country:
| UK |
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| Telephone Number:
| +44 (0)115 9516377 |
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Fax Number:
| +44 (0)115 9516334 |
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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:
| genetic_modification_design |
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Number of Slides: | 12 |
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| Experimental Parameters:
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parameter | gene_knock_out |
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Quality Control Measures Taken:
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References:
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reference | Dawson J, Sozen E, Vizir I, Van Waeyenberge S, Wilson ZA and Muligan BJ (1999): Characterization and genetic mapping of a mutation (ms35) which prevents anther dehiscence in Arabidopsis thaliana by affecting secondary wall thickening in endothecium. New Phytologist, 144: 213-222 |
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Slides in this Experiment
Hybridisation Set: Song: Analysis of anther development by identifiying downstream genes controlled by MS35/MYB26_genome
Slide: Song_1-1_Ler_3.4.5_Rep1_ATH1 | | |
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Tissue:
| anther and filament |
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Other Information:
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Sample description | Ler.gl and ms35.gl anther and filaments were collected from three developmental stages separately, PM I, bicellular and PM II. The use of the gl marker enabled a prompt identification of the ms35 mutant amongst the segregation population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation. |
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Protocols for BioSource 1 |
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Slide: Song_1-2_Ler_6.7_Rep1_ATH1 | | |
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Tissue:
| anther and filament |
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Other Information:
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Sample description | Ler.gl and ms35.gl anther and filaments were collected from three developmental stages separately, PM I, bicellular and PM II. The use of the gl marker enabled a prompt identification of the ms35 mutant amongst the segregation population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation. |
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Protocols for BioSource 1 |
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Slide: Song_1-3_Ler_8.9.10_Rep1_ATH1 | | |
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Tissue:
| anther and filament |
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Other Information:
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Sample description | Ler.gl and ms35.gl anther and filaments were collected from three developmental stages separately, PM I, bicellular and PM II. The use of the gl marker enabled a prompt identification of the ms35 mutant amongst the segregation population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation. |
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Protocols for BioSource 1 |
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Slide: Song_1-4_MS35_3.4.5_Rep1_ATH1 | | |
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Tissue:
| anther and filament |
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Other Information:
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Sample description | Ler.gl and ms35.gl anther and filaments were collected from three developmental stages separately, PM I, bicellular and PM II. The use of the gl marker enabled a prompt identification of the ms35 mutant amongst the segregation population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation. |
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Protocols for BioSource 1 |
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Slide: Song_1-5_MS35_6.7_Rep1_ATH1 | | |
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Tissue:
| anther and filament |
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Other Information:
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Sample description | Ler.gl and ms35.gl anther and filaments were collected from three developmental stages separately, PM I, bicellular and PM II. The use of the gl marker enabled a prompt identification of the ms35 mutant amongst the segregation population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation. |
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Protocols for BioSource 1 |
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Slide: Song_1-6_MS35_8.9.10_Rep1_ATH1 | | |
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Tissue:
| anther and filament |
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Other Information:
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Sample description | Ler.gl and ms35.gl anther and filaments were collected from three developmental stages separately, PM I, bicellular and PM II. The use of the gl marker enabled a prompt identification of the ms35 mutant amongst the segregation population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation. |
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Protocols for BioSource 1 |
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Slide: Song_1-7_Ler_3.4.5_Rep2_ATH1 | | |
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Tissue:
| anther and filament |
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Other Information:
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Sample description | Ler.gl and ms35.gl anther and filaments were collected from three developmental stages separately, PM I, bicellular and PM II. The use of the gl marker enabled a prompt identification of the ms35 mutant amongst the segregation population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation. |
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Protocols for BioSource 1 |
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Slide: Song_1-8_Ler_6.7_Rep2_ATH1 | | |
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Tissue:
| anther and filament |
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Other Information:
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Sample description | Ler.gl and ms35.gl anther and filaments were collected from three developmental stages separately, PM I, bicellular and PM II. The use of the gl marker enabled a prompt identification of the ms35 mutant amongst the segregation population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation. |
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Protocols for BioSource 1 |
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Slide: Song_1-9_Ler_8.9.10_Rep2_ATH1 | | |
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Tissue:
| anther and filament |
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Other Information:
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Sample description | Ler.gl and ms35.gl anther and filaments were collected from three developmental stages separately, PM I, bicellular and PM II. The use of the gl marker enabled a prompt identification of the ms35 mutant amongst the segregation population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation. |
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Protocols for BioSource 1 |
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Slide: Song_1-10_MS35_3.4.5_Rep2_ATH1 | | |
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Tissue:
| anther and filament |
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Other Information:
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Sample description | Ler.gl and ms35.gl anther and filaments were collected from three developmental stages separately, PM I, bicellular and PM II. The use of the gl marker enabled a prompt identification of the ms35 mutant amongst the segregation population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation. |
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Protocols for BioSource 1 |
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Slide: Song_1-11_MS35_6.7_Rep2_ATH1 | | |
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Tissue:
| anther and filament |
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Other Information:
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Sample description | Ler.gl and ms35.gl anther and filaments were collected from three developmental stages separately, PM I, bicellular and PM II. The use of the gl marker enabled a prompt identification of the ms35 mutant amongst the segregation population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation. |
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Protocols for BioSource 1 |
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Slide: Song_1-12_MS35_8.9.10_Rep2_ATH1 | | |
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Tissue:
| anther and filament |
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Other Information:
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Sample description | Ler.gl and ms35.gl anther and filaments were collected from three developmental stages separately, PM I, bicellular and PM II. The use of the gl marker enabled a prompt identification of the ms35 mutant amongst the segregation population. The wild-type control line was chosen to be Ler.gl which ensures that the changes of gene expression are not due to the gl mutation. |
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Protocols for BioSource 1 |
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Problems? Comments? Suggestions? Contact the Affymetrix Team