Summary of Study ST002571
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001658. The data can be accessed directly via it's Project DOI: 10.21228/M8CX2C This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
Study ID | ST002571 |
Study Title | Quantification of cytokinins in ap1 cal AP1-GR and drmy1 ap1 cal AP1-GR inflorescences using LC-MS |
Study Type | Quantification using mass spectrometry |
Study Summary | Robustness is the invariant development of phenotype despite environmental changes and genetic perturbations. In the Arabidopsis flower bud, four sepals initiate at robust positions and times and grow to equal size to enclose and protect the inner floral organs. We previously characterized the mutant development related myb-like1 (drmy1), where 3-5 sepals initiate at irregular positions and variable times and grow to different sizes, compromising their protective function. This loss of robustness was caused by a uniform increase in cytokinin signaling, as revealed by the TCS::GFP reporter, in the floral meristem before sepal initiation. We hypothesized that the increase in cytokinin signaling in drmy1 was due to an increase in the level of cytokinins. To test this idea, we extracted cytokinins from induced inflorescences of wild-type (5 bio-reps) and drmy1 (6 bio-reps) in ap1 cal AP1-GR background. We measured the level of three cytokinin bases, trans-Zeatin (tZ), cis-Zeatin (cZ), and isopentenyladenine (iP), and their corresponding nucleosides (tZR, cZR, and iPR), using liquid chromatography-mass spectrometry. We found that there was no statistically significant differences in cytokinin levels between these genotypes, indicating that the increase in cytokinin signaling in the drmy1 floral meristem is not due to increased cytokinin levels. |
Institute | Cornell University |
Department | Plant Biology Section |
Laboratory | Roeder Lab |
Last Name | Kong |
First Name | Shuyao |
Address | 239 Weill Hall, 526 Campus Road, Ithaca, NY 14853 |
sk3245@cornell.edu | |
Phone | 6072629684 |
Submit Date | 2023-04-20 |
Num Groups | 2 |
Total Subjects | 11 |
Raw Data Available | Yes |
Raw Data File Type(s) | raw(Thermo) |
Analysis Type Detail | LC-MS |
Release Date | 2023-05-11 |
Release Version | 1 |
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Sample Preparation:
Sampleprep ID: | SP002678 |
Sampleprep Summary: | Samples were ground in liquid nitrogen and twice extracted in methanol : water : formic acid (15:4:1). 200 pg of BAP per sample was added as an internal control. Extracts were centrifuged at 14,650 rpm in -4°C for 30 min, and supernatant was evaporated of methanol and reconstituted in 1% (v/v) acetic acid. Samples were passed through an Oasis MCX SPE column (Waters 186000252), washed with 1% acetic acid, washed with methanol, and eluted with 0.35 M ammonia in 70% methanol. Eluents were evaporated to complete dryness, reconstituted in 5% acetonitrile, and sent for LC-MS. |
Processing Storage Conditions: | -80℃ |
Extraction Method: | Modified Bieleski buffer, methanol : water : formic acid (15:4:1) |
Extract Enrichment: | An Oasis MCX SPE column (Waters 186000252) was used to enrich for cytokinins |
Extract Storage: | -80℃ |
Sample Resuspension: | 5% acetonitrile |
Sample Spiking: | 200 pg of BAP per sample was added as an internal control |