140515H2

Property Value
BioDare ID 3188
Author Sarah Hodge
Institution MPIMP - UEDIN
License

Description

ZT0 lights remain switched off until ZT3. At ZT3 light comes on at 90 umolm-2s-1. Dusk is lights off at ZT 12. Dark dawn cycle is for 4 days then continuous light (first day with dark dawn) for 3 days. ie 9L:15D x4 LLx3.

Purpose

Control "dark" dawn experiment for double dawn experiment based on Virginie and Thiago's design (Stitt Lab).

Comments

Partner experiment: 140515H1

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 2)

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 -- -- --

3D:9L:12Dx3 LLx3

Growth on Agar 1.2% MS 0.5 for 7.0 days (3D:9L:12Dx3 LLx3). 4 days of 3 hours dark dawn, 9 hours of bright light 90 umolm-2-s-1, 12 hours dark followed by 3 days constant light, with dark dawn conditions on first dawn. 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 3:00 9:00 red LED 55
diurnal light 1 24 3:00 21:00 red LED 55
constant light 2 24 0:00 24:00 red LED 55
diurnal light 4 24 3:00 9:00 blue LED 35
diurnal light 1 24 3:00 21:00 blue LED 35
constant light 2 24 0:00 24:00 blue LED 35

Temperature

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

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