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A nuclear-encoded ClpP subunit of the chloroplast ATP-dependent Clp protease is essential for early development in Arabidopsis thaliana

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Abstract

ClpP4 is a nuclear-encoded plastid protein that functions as a proteolytic subunit of the ATP-dependent Clp protease of higher plants. Given the lack of viable clpP4 knockout mutants, antisense clpP4 repression lines were prepared to study the functional importance of ClpP4 in Arabidopsis thaliana. Screening of transformants revealed viable lines with up to 90% loss of wild type levels of ClpP4 protein, while those with > 90% were severely bleached and strongly retarded in vegetative growth, failing to reach reproductive maturity. Of the viable antisense plants, repression of clpP4 expression produced a pleiotropic phenotype, of which slow growth and leaf variegation were most prominent. Chlorosis was most severe in younger leaves, with the affected regions localized around the mid-vein and exhibiting impaired chloroplast development and mesophyll cell differentiation. Chlorosis lessened during leaf expansion until all had regained the wild type appearance upon maturity. This change in phenotype correlated with the developmental expression of ClpP4 in the wild type, in which ClpP4 was less abundant in mature leaves due to post-transcriptional/translational regulation. Repression of ClpP4 caused a concomitant down-regulation of other nuclear-encoded ClpP paralogs in the antisense lines, but no change in other chloroplast-localized Clp proteins. Greening of the young chlorotic antisense plants upon maturation was accelerated by increased light, either by longer photoperiod or by higher growth irradiance; conditions that both raised levels of ClpP4 in wild type leaves. In contrast, shift to low growth irradiance decreased the relative amount of ClpP4 in wild type leaves, and caused newly developed leaves of fully greened antisense lines to regain the chlorotic phenotype.

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Abbreviations

CaMV:

Cauliflower mosaic virus

HSP:

Heat shock protein

J PSII :

PSII electron transport

Kan:

Kanamycin

NPQ:

Nonphotochemical quenching

q P :

Photochemical quenching

ΦPSII :

Photosynthetic yield of PSII

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Acknowledgements

This work was supported by grants to A.K.C. from The Swedish Research Council for Environment, Agricultural Science and Spatial Planning (Formas) and the Swedish Foundation for International Cooperation in Research and Higher Education (STINT). T.M.M. would like to acknowledge NSERC for an oversea postgraduate scholarship.

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Authors and Affiliations

  1. Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, 901 87, Umea, Sweden

    Bo Zheng & Vaughan Hurry

  2. Plant Molecular Biology, Department of Plant and Environmental Sciences, Göteborg University, PO Box 461, 405 30, Goteborg, Sweden

    Tara M. MacDonald & Adrian K. Clarke

  3. Finnish Forest Research Institute, Joensuu Research Center, 801 01 , Joensuu, Finland

    Sirkka Sutinen

Authors
  1. Bo Zheng
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  2. Tara M. MacDonald
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  3. Sirkka Sutinen
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  4. Vaughan Hurry
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  5. Adrian K. Clarke
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Correspondence to Adrian K. Clarke.

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Zheng, B., MacDonald, T.M., Sutinen, S. et al. A nuclear-encoded ClpP subunit of the chloroplast ATP-dependent Clp protease is essential for early development in Arabidopsis thaliana . Planta 224, 1103–1115 (2006). https://doi.org/10.1007/s00425-006-0292-2

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