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Chloroplast Phenomics: Systematic Phenotypic Screening of Chloroplast Protein Mutants in Arabidopsis

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Chloroplast Research in Arabidopsis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 775))

Abstract

As part of a project to analyze chloroplast functional networks systematically, we have subjected mutants in >3,200 nuclear genes predicted to encode chloroplast-targeted proteins in Arabidopsis thaliana (http://www.plastid.msu.edu) to parallel phenotypic assays. Detailed methods are presented for the various assays being used in this project to study chloroplast biology. These include morphological analysis of plants, chloroplasts, and seeds using controlled vocabulary. Metabolites synthesized in the chloroplast such as starch, amino acids, and fatty acids are analyzed in groups according to their chemical properties. As an indicator for the relative composition of seed storage oil and proteins, the carbon and nitrogen contents are determined by an elemental analyzer. The methods in this chapter describe how the assays are configured to run in relatively high throughput, maximizing data quality.

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Acknowledgments

The authors thank Kathleen M. Imre for configuring the chlorophyll fluorescence assay and Imad Ajjawi for configuring the fatty acid assay. We are grateful to the many project members who contributed to the establishment and refinement of these protocols, including the many undergraduate students involved in the project. This work was supported by the US National Science Foundation 2010 Project Grants MCB-0519740 and DBI-0619489 for LC-MS equipment.

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Authors and Affiliations

  1. Department of Biological Sciences, Western Michigan University, Kalamazoo, MI, USA

    Yan Lu

  2. Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, USA

    Linda J. Savage

  3. Department of Biochemistry and Molecular Biology, and Department of Plant Biology, Michigan State University, East Lansing, MI, USA

    Robert L. Last

Authors
  1. Yan Lu
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  2. Linda J. Savage
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Correspondence to Yan Lu .

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Editors and Affiliations

  1. , Department of Biology, University of Leicester, University Road, Leicester, LE1 7RH, United Kingdom

    R. Paul Jarvis

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Lu, Y., Savage, L.J., Last, R.L. (2011). Chloroplast Phenomics: Systematic Phenotypic Screening of Chloroplast Protein Mutants in Arabidopsis. In: Jarvis, R. (eds) Chloroplast Research in Arabidopsis. Methods in Molecular Biology, vol 775. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-237-3_9

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  • DOI: https://doi.org/10.1007/978-1-61779-237-3_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-236-6

  • Online ISBN: 978-1-61779-237-3

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