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By Bill Davey, Clive Mathews

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Methods utilizing these properties were first developed four decades ago [8] and have since then been used by others with modifications [9–11]. With the use of commonly available programmable microplate readers, quantitative and automatic time course data of chloroplast movements can be obtained with ease. In this chapter, we describe this method, including the setup of the measuring equipment, preparation of samples and data analysis. 2 Material 1. Plate reader: Should be able to be programmed to eject and insert a 96-well plate at any given time to measure absorbance of 660 nm and 800 nm.

B) The plant growth parameters can be measured in the original photos by hand using ImageJ, Adobe Photoshop or other standard image analysis software. (c) Custom image enhancement scripts can digitize the images making it easier to analyze the whole time-lapse series using specific root tip tracking software 70 μmol m−2 s−1 of light. The room was kept between 22 and 24 °C and had a humidity between 50 and 60 % in front of the camera (see Note 7). 6. Rotate plates 90° and set the camera to photograph every 30 min for 12 h (see Note 8 and Fig.

Nature 482(7383):103–106 11. Brooks T, Miller N, Spalding E (2010) Plasticity of Arabidopsis root Gravitropism throughout a multidimensional condition space quantified by automated image analysis. Plant Physiol 152(1):206–216 12. Wells D, French A, Naeem A, Ishaq O, Traini R, Hijazi H, Bennett M, Pridmore T (2012) Recovering the dynamics of root growth and development using novel image acquisition and analysis methods. Philos Trans R Soc B: Biol Sci 367(1595):1517–1524 13. Sangari A, Ohme-Takagi M, Mitsuda N, Assadi A (2012) Automated imaging & highthroughput phenotyping, massively parallel image acquisition, analysis and modeling: mapping plant genotype-phenotype relation.

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