Arabidopsis thaliana Col-0 seeds were sown in SunGrow Professional growing potting mix in 3.5-inch square pots, stratified for one day at 4°C in darkness, then moved to Conviron A1000 growth chamber with settings of 14 hour day, 23°C, 70 μmol light. After two weeks, the pots were thinned to 4 plants per pot. After four weeks, the pots were moved to a growth room with settings of 12 hour day, 23°C.
A1000 growth chamber
Manufactured by Conviron
Sourced in Canada
The A1000 growth chamber is a controlled environment chamber designed for plant growth and research applications. It provides a precisely regulated temperature, humidity, and lighting environment to support the cultivation of various plant species. The chamber's core function is to create and maintain a stable and consistent growth environment for scientific experiments and plant research.
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6 protocols using a1000 growth chamber
1
Cultivation of Arabidopsis thaliana
2
Arabidopsis Responses to Iron Status
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Arabidopsis thaliana ecotype Columbia (Col-0) was used as the wild type. The clf mutant line used was clf-29 (SALK_21003) from the Arabidopsis Biological Resource Center. Arabidopsis seeds were surface-sterilized and plated on B5 media. After stratification, seeds were grown at 22°C under a 16/8 h light/dark cycle in a Conviron A1000 growth chamber. For iron treatments, plants were transferred to iron deficient media (-Fe), Murashige and Skoog (MS) medium with no iron (Caisson MSP33) supplemented with 300 μM ferrozine, or Fe sufficient media (+Fe), MS medium with 100 μM FeNa-EDTA (Caisson MSP34) for 3 days for both gene expression analysis and chromatin immunoprecipitation.
3
X-ray CT Imaging of Plant Roots
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A17 WT or cra2-11 were grown in sieved (< 2 mm) sandy loam soil uniformly packed to a bulk density of 1.1 mg m À3 in a 68 mm (diameter) 9 160 mm (height) cylindrical column made from high density polyethylene (Mairhofer et al., 2017) for 21 d in a Conviron A1000 growth chamber at 22°C, 60% humidity with a 16 h photoperiod. After 14 d or 21 d growth, each column was scanned using a GE v|tome|x M 240 kV X-ray CT system at the Hounsfield Facility, University of Nottingham (UK). Scans were made in 'fast mode', collecting a single radiograph image for each of the 2400 angular projections over a 360°rotation of the sample at spatial resolution of 40 µm. Scans were made in three sections to obtain the full length of the column/soil depth. Data were reconstructed and subjected to manual root segmentation techniques to digitally separate the roots from the soil. Extracted RSAs were quantified using ROOTH software (Mairhofer et al., 2017) for total root length, branch structure and LR angle.
4
Characterizing fpn3 T-DNA Insertion Lines
5
Brachypodium distachyon Growth and Imaging
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Dry seed of B. distachyon accession Bd21-3 was imbibed and stratified in a wet paper towel at 6°C for sixteen days. Seeds were then sown towards one edge of 10 cm pots containing potting mix (#2; Conrad Fafard Inc., Agawa, MA, USA). Half of the soil was covered with 0.508 mm thick infrared absorbing paper followed by 0.254 mm infrared light absorbing paper (Edmund Optics; Barrington, NJ, USA). Plants were cultivated and imaged in a Percival model CU36L6 Growth Chamber (Percival Scientific, Perry, IA, USA). Light conditions were 62 to 74 µmol of photons⋅m−2⋅s−1 unless otherwise indicated as bright light, which was 300 to 340 µmol of photons⋅m−2⋅s−1 in a Conviron model A1000 Growth Chamber (Conviron, Winnipeg, MB, Canada). The hot cold temperature parameters correspond to 28±0.6°C and 12±0.6°C, respectively. Percival conditions were monitored and recorded using a HOBO U12 Data Logger (Onset Computer Corporation, Bourne, MA, USA).
6
Characterizing fpn3 T-DNA Insertion Lines
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Two independent alleles of fpn3 T-DNA insertion lines, fpn3-1 (SALK_034189) and fpn3-2 (SALK_009286), were obtained from the Salk collection (Alonso et al., 2003) . RT-qPCR verified that FPN3 expression was drastically reduced in these lines (Supplementary Figure 3). Arabidopsis plants were grown at 22 °C under a 16/8-hour light/dark cycle in the Conviron A1000 growth chamber.
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