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Inhibition of lycopene cyclase results in accumulation of chlorophyll precursors

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Abstract

Free porphyrins and their magnesium complexes, including chlorophylls, are potent photo-sensitizers. Plants usually accumulate these compounds bound to proteins together with protective compounds like carotenoids. Besides their protective role, carotenoids can play a structural role in these complexes. To analyze the effect of impaired carotenogenesis on plastid membranes we applied to barley seedlings the bleaching herbicide 2-(4-chlorophenylthio)triethylamine (CPTA) as a specific inhibitor for the cyclization of lycopene. To avoid interference with photo-oxidation, the essential experiments were performed on seedlings grown in darkness. While the amount of total carotenoids decreased, we found accumulation of more δ-carotene than lycopene in darkness clearly showing that CPTA inhibits the lycopene β-cyclase more effectively than the lycopene ε-cyclase. The CPTA treatment resulted in accumulation of non-photoactive protochlorophyllide a; the amount of photoactive protochlorophyllide and NADPH:protochlorophyllide oxidoreductase remained constant. Further, the level of Mg protophorphyrin and its monomethyl ester increased to an extent similar to that obtained by application of 5-aminolevulinic acid (ALA). The perturbation of the ultrastructure of etioplast inner membranes, observed after CPTA-treatment, was not found after ALA-treatment; this excluded the accumulated tetrapyrroles as responsible for the perturbation. By contrast, the down-regulation of Lhcb and RbcS genes found after CPTA-treatment was compatible with the presumed role of Mg protophorphyrin as “plastid signal” for regulation of nuclear gene expression. Possible mechanisms for enhancement of tetrapyrrole accumulation by non-cyclic carotenoids are discussed.

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Abbreviations

ALA:

5-Aminolevulinic acid

CPTA:

2-(4-chlorophenylthio) triethylamine

MgPPIX:

Magnesium protoporphyrin IX

MgPPIXMe:

Magnesium protoporphyrin IX monomethyl ester

Pchlide:

Protochlorophyllide a

POR:

NADPH:protochlorophyllide oxidoreductase

PPIX:

Protoporphyrin IX

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Acknowledgments

We thank Dr. A. Trebst (Universität Bochum, Germany for a gift of CPTA, Dr. G. Sandmann (Universität Frankfurt, Germany) for a sample of δ-carotene, Dr. S. Pancaldi (University of Ferrara, Italy) for a gift of samples of neoxanthin, lutein, violaxanthin and antheraxanthin and Dr. M. Ryberg (University of Göteborg, Sweden) for an antiserum raised against POR. The research was supported by the Deutsche Forschungsgemeinschaft, Bonn, and by the Ministero dell’Istruzione, dell’Università e della Ricerca (MIUR) Italiano.

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Authors and Affiliations

  1. Dipartimento di Biologia, Università di Padova, Via U. Bassi 58/B, 35131, Padova, Italy

    Nicoletta La Rocca & Nicoletta Rascio

  2. Department Biologie I (Botanik), Universität München, Menzinger Str. 67, 80638, Munich, Germany

    Ulrike Oster & Wolfhart Rüdiger

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  1. Nicoletta La Rocca
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  2. Nicoletta Rascio
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  3. Ulrike Oster
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  4. Wolfhart Rüdiger
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Correspondence to Wolfhart Rüdiger.

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La Rocca, N., Rascio, N., Oster, U. et al. Inhibition of lycopene cyclase results in accumulation of chlorophyll precursors. Planta 225, 1019–1029 (2007). https://doi.org/10.1007/s00425-006-0409-7

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