140515H1

Property Value
BioDare ID 3187
Author Sarah Hodge
Institution MPIMP - UEDIN
License

Description

At ZT0 dawn is dim light (6umolm-2s-1) for 3 hours. At ZT3 light steps up to 90 umolm-2s-1. Dusk is lights off at ZT 12. Double dawn cycle is for 4 days then continuous light (first day with double dawn) for 3 days. ie 12L:12D x4 LLx3.

Purpose

Double dawn experiment based on Virginie and Thiago's design (Stitt Lab).

Comments

Pedro is not able to record changes in intensity during an experiment, so please refer to the description & experimental conditions comment for true experimental conditions. Partner experiment: 140515H2

Sample preparation

Plants sprayed with luciferin on day 6 of growth conditions, returned to entrainment conditions for a further 24hours, they were transferred to experimental conditions at ZT4 on d7. Imaging was started on day 8.

Measurement

Protocol

Luciferase luminescence (HAM 1)

Seedlings were treated on 6th day with Luciferin. They were then returned to entrainment condition for a further 24h (due to imaging set up problems). Seedlings were transferred to Experimental conditions at ZT4 on d7. Imaging was started on d8.

Imaging script started 5 minutes before ZT0 55mins wait, 5mins lights off, 30mins capture

Experimental conditions

12L:12D

Growth on Agar 1.2% MS 0.5 for 7.0 days (12L:12D). Plants were entrained at light intensity 90 umolm-2s-1 to correspond to the light intensity of the experiment. The plants were entrained for 1 day more than standard conditions due problems with imaging set up.

Light

Type Duration (days) Cycle (h) Start Duration Spectrum Source Intensity
diurnal light 7 24 0:00 12:00 white tube 90

Temperature

Type Duration (days) Cycle (h) Base (°C) Warm (°C) Warm start Warm duration
constant temperature 7 24 22 -- -- --

3ShL:9L:12Dx4 LLx3

Growth on Agar 1.2% MS 0.5 for 7.0 days (3ShL:9L:12Dx4 LLx3). 4 days of 3 hours dim light (6 umolm-2s-1), 9 hours of bright light (90 umolm-2-s), 12 hours dark followed by 3 days constant light, with shady conditions on first dawn. The first dawn starts at ZT0 The light intensities are controlled by the arduino. the plates were transferred from entrainment conditions to experimental conditions at ZT4 on day 7.

Light

Type Duration (days) Cycle (h) Start Duration Spectrum Source Intensity
diurnal light 4 24 0:00 12:00 blue LED 35
constant light 3 24 0:00 24:00 blue LED 35
diurnal light 4 24 0:00 12:00 red LED 55
constant light 3 24 0:00 24:00 red LED 55

Temperature

Type Duration (days) Cycle (h) Base (°C) Warm (°C) Warm start Warm duration
constant temperature 7 24 22 -- -- --

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