| Property | Value |
|---|---|
| BioDare ID | 3648 |
| Author | Victoria Hibberd |
| Institution | University of Edinburgh |
| License | CC_BY |
If periods change during development, then our genetic screens for preiod-altering clock mutants might recover developmental mutants. Leaf movement assays suggestted that period might change. Ten-day data collections in the TopCount were intended to test for such period changes using luciferase. The lhy cca1 mutant was of particular interest, as it had a very short period to start with - was this perhaps rescued in older plants? In fact, ultradian rhythms were revealed.. Replicated in Long #2 experiment, Oct 2003.
Test for period changes during seedlling growth in the TopCount, in WT and cca lhy double mutants
Data collection by Victoria Hibberd ca. September 2003; files found by Andrew Millar in 2012, reanalysed on sabbatical in 2015. Note only 4 plants of line 213 in this replicate. Line 313 replaces it in #2 replicate.
Luciferin pipetted on to seedling, 24h before counting began. Plate layout: each genotype occupied two full rows of 12 wells per row (give or take a few dead seedlings).
Luciferase luminescence (Top count 1)
Each plate held 5 mins in dark before counting started. Counting with 2-detector TopCount took ca. 2.8 mins per plate. Each plate counted every 29.01 minutes (XL trendline is actually 29.8 mins, but no matter).
Growth on Agar 1.5% MS 1 3% Sucrose for 6.0 days (12L:12D WL). Standard Warwick procedure would be described in (e.g.) Hall et al. TIC paper Plant Cell 2003.
| Type | Duration (days) | Cycle (h) | Start | Duration | Spectrum | Source | Intensity |
|---|---|---|---|---|---|---|---|
| diurnal light | 6 | 24 | 0:00 | 12:00 | white | tube | 80 |
| Type | Duration (days) | Cycle (h) | Base (°C) | Warm (°C) | Warm start | Warm duration |
|---|---|---|---|---|---|---|
| constant temperature | 6 | 24 | 22 | -- | -- | -- |
Growth on Agar 1.5% MS 1 3% Sucrose for 11.0 days (constant R+B). again, standard Warwick procedure as described in Hall et al. Plant Cell 2003. However, light levels in the black wells in the TopCount were very low. 2uE each of R+B is the maximum. R and B LEDs were in alternating rows of colour, next to the stackers. LEDs at each end of the plate (i.e. next to well columns 1 and 12) and would be low (1-2 micromol/m2/sec) and uneven (ca. half in rows 6,7) despite the mirror plates.
| Type | Duration (days) | Cycle (h) | Start | Duration | Spectrum | Source | Intensity |
|---|---|---|---|---|---|---|---|
| constant light | 11 | 24 | 0:00 | 24:00 | red | LED | 2 |
| constant light | 11 | 24 | 0:00 | 24:00 | blue | LED | 2 |
| Type | Duration (days) | Cycle (h) | Base (°C) | Warm (°C) | Warm start | Warm duration |
|---|---|---|---|---|---|---|
| constant temperature | 11 | 24 | 22 | -- | -- | -- |