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Arabidopsis light-dependent protochlorophyllide oxidoreductase A (PORA) is essential for normal plant growth and development

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Abstract

During skotomorphogenesis in angiosperms, NADPH:protochlorophyllide oxidoreductase (POR) forms an aggregate of photolabile NADPH-POR-protochlorophyllide (Pchlide) ternary complexes localized to the prolamellar bodies within etioplasts. During photomorphogenesis, POR catalyzes the light-dependent reduction of Pchlide a to chlorophyllide (Chlide) a, which is subsequently converted to chlorophyll (Chl). In Arabidopsis there are three structurally related POR genes, denoted PORA, PORB and PORC. The PORA and PORB proteins accumulate during skotomorphogenesis. During illumination, PORA is only transiently expressed, whereas PORB and PORC persist and are responsible for bulk Chl synthesis throughout plant development. Here we have tested whether PORA is important for skotomorphogenesis by assisting in etioplast development, and normal photomorphogenic development. Using reverse genetic approaches, we have identified the porA-1 null mutant, which contains an insertion of the maize Dissociation transposable element in the PORA gene. Additionally, we have characterized PORA RNAi lines. The porA-1 and PORA RNAi lines display severe photoautotrophic growth defects, which can be partially rescued on sucrose-supplemented growth media. Elimination of PORA during skotomorphogenesis results in reductions in the volume and frequency of prolamellar bodies, and in photoactive Pchlide conversion. The porA-1 mutant characterization thus establishes a quantitative requirement for PORA in etioplast development by demonstrating significant membrane ultrastructural and biochemical defects, in addition to suggesting PORA-specific functions in photomorphogenesis and plant development.

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Acknowledgments

We thank Rosario Barbieri for advice, Rebecca Lamb for her comments, Kathy Wolken at The Ohio State University Campus Microscopy and Imaging Facility for assistance with the electron microscopy, Dick Sayre for providing access to the fluorescence spectrophotometer, and the AGRIKOLA consortium (http://www.agrikola.org), the Nottingham Arabidopsis Stock Centre (Nottingham, UK) and the Arabidopsis Biological Resource Center (Columbus, OH, USA) for the generous donation of seed lines. This work was financially supported by The Ohio State University and the National Science Foundation (IOB #0450114).

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Authors and Affiliations

  1. Department of Plant Cellular and Molecular Biology, The Ohio State University, 500 Aronoff Laboratory, 318 West 12th Ave., Columbus, OH, 43210-1293, USA

    Troy Paddock, Daniel Lima & Gregory A. Armstrong

  2. Department of Entomology, Ohio Agricultural Research and Development Center, 1060 Madison Ave., Wooster, OH, 44691, USA

    Mary E. Mason

  3. Boyce Thompson Institute for Plant Research, Tower Road, Ithaca, NY, 14853-1801, USA

    Klaus Apel

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  1. Troy Paddock
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Correspondence to Troy Paddock.

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Paddock, T., Lima, D., Mason, M.E. et al. Arabidopsis light-dependent protochlorophyllide oxidoreductase A (PORA) is essential for normal plant growth and development. Plant Mol Biol 78, 447–460 (2012). https://doi.org/10.1007/s11103-012-9873-6

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