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Experiment: Genes Involved in Meristemoid Cell Fate
Experiment Description
NASCArrays Experiment Reference Number: NASCARRAYS-388
The objective of this experiment is to identify genes that are regulated by the LRR-receptor-like protein TOO MANY MOUTHS (TMM) to control stomatal precursor (meristemoid) cell fate and behavior. An interesting aspect of the tmm phenotype is that mutant plants overproduce stomata on leaves, but lack stomata entirely on inflorescence stems and in other locations on the plant. Despite the failure to produce mature stomata, mutant stems produce presumptive stomatal precursors through asymmetric divisions. This suggests that normal TMM signaling is required to maintain meristemoid cell fate but not to execute formative asymmetric divisions. Because stem stomata develop in a morphogenetic wave from tip to base, it is possible to harvest tissues that contain meristemoids but not guard mother cells or mature stomata. This provides an opportunity to compare the gene expression profiles of tissues with meristemoids that maintain their cellular identity and ultimately produce stomata (Col gl1), to tissues in which meristemoids form but rapidly lose their cellular identity due to a failure in TMM signaling (tmm-1). Because guard mother cells and stomata are not present in the tissue harvested, genes that change in abundance during these developmental stages will be filtered out. Ultimately we hope to reveal genes that operate downstream of TMM to confer meristemoid cell fate or to provide 'niche factors' suitable for their development into stomata.
For analysis, approximately 90 developing inflorescence stem tips were collected and flower buds and meristems removed prior to flash freezing for each replicate sample. The region harvested had been previously determined to contain developing meristemoids but not stomata or guard mother cells by microscopic observation. An identical region was harvested from tmm-1 mutant plants grown at the same time, in the same flat.
About the ExperimenterName: | Jeanette Nadeau |
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Head of Lab Name: | Jeanette Nadeau |
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Lab:
| Nadeau lab |
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Institute:
| University of Central Florida |
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Address: | Department of Biology 4000 Central Florida Blvd. Orlando, FL
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Postcode:
| 32816-2368 |
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Country:
| USA |
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| Telephone Number:
| 407-823-1663 |
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Fax Number:
| 407-823-5769 |
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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: | 4 |
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| Experimental Parameters:
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| gene_knock_out |
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Quality Control Measures Taken:
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References:
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| Other Information:
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Slides in this Experiment
Hybridisation Set: Nadeau: Genome set
Slide: Nadeau_1-1_tmm1-inflorescence-tips-pool1_Rep1_ATH1 | | |
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Tissue:
| inflorescence tip |
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Diseased:
| Normal |
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Additional Organism Information:
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separation-technique | The inflorescence stem containing immature epidermis was harvested as follows: open flowers and buds were rapidly removed from the developing inflorescence stem at the base of the pedicels using a dissecting microscope and fine point forceps. Meristems were then removed from the tip. Tips of the inflorescence stem were removed from the plant and flash frozen in microcentrifuge tubes. Each sample was composed of pooled tips from ~90 inflorescences. Prior to harvesting stems for RNA, approximately 20 inflorescence tips were examined with a microscope to identify the region of the stem that consistently contained developing stomatal meristemoids in the epidermis, but few guard mother cells or mature stomata. These stems were not included in the RNA sample |
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sampledescription | Bolting Col-3 gl1 plants |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Nadeau_1-2_Col1g11-inflorescence-tips-pool1_Rep1_ATH1 | | |
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Tissue:
| inflorescence tip |
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Diseased:
| Normal |
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Additional Organism Information:
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separation-technique | The inflorescence stem containing immature epidermis was harvested as follows: open flowers and buds were rapidly removed from the developing inflorescence stem at the base of the pedicels using a dissecting microscope and fine point forceps. Meristems were then removed from the tip. Tips of the inflorescence stem were removed from the plant and flash frozen in microcentrifuge tubes. Each sample was composed of pooled tips from ~90 inflorescences. Prior to harvesting stems for RNA, approximately 20 inflorescence tips were examined with a microscope to identify the region of the stem that consistently contained developing stomatal meristemoids in the epidermis, but few guard mother cells or mature stomata. These stems were not included in the RNA sample |
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sampledescription | Bolting Col-3 gl1 plants |
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Other Information:
| Lehle Seeds |
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Protocols for BioSource 1 |
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Slide: Nadeau_1-3_tmm1-inflorescence-tips-pool2_Rep1_ATH1 | | |
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Tissue:
| inflorescence tip |
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Diseased:
| Normal |
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Additional Organism Information:
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separation-technique | The inflorescence stem containing immature epidermis was harvested as follows: open flowers and buds were rapidly removed from the developing inflorescence stem at the base of the pedicels using a dissecting microscope and fine point forceps. Meristems were then removed from the tip. Tips of the inflorescence stem were removed from the plant and flash frozen in microcentrifuge tubes. Each sample was composed of pooled tips from ~90 inflorescences. Prior to harvesting stems for RNA, approximately 20 inflorescence tips were examined with a microscope to identify the region of the stem that consistently contained developing stomatal meristemoids in the epidermis, but few guard mother cells or mature stomata. These stems were not included in the RNA sample |
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sampledescription | Bolting Col-3 gl1 plants |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Nadeau_1-4_Col1g11-inflorescence-tips-pool2_Rep1_ATH1 | | |
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Tissue:
| inflorescence tip |
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Diseased:
| Normal |
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Additional Organism Information:
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separation-technique | The inflorescence stem containing immature epidermis was harvested as follows: open flowers and buds were rapidly removed from the developing inflorescence stem at the base of the pedicels using a dissecting microscope and fine point forceps. Meristems were then removed from the tip. Tips of the inflorescence stem were removed from the plant and flash frozen in microcentrifuge tubes. Each sample was composed of pooled tips from ~90 inflorescences. Prior to harvesting stems for RNA, approximately 20 inflorescence tips were examined with a microscope to identify the region of the stem that consistently contained developing stomatal meristemoids in the epidermis, but few guard mother cells or mature stomata. These stems were not included in the RNA sample |
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sampledescription | Bolting Col-3 gl1 plants |
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Other Information:
| Lehle Seeds |
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Protocols for BioSource 1 |
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