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Senescence in wheat leaves: is a cysteine endopeptidase involved in the degradation of the large subunit of Rubisco?

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Abstract

In wheat (Triticum aestivum L.), leaf senescence can be initiated by different factors. Depending on the plant system (intact plants or detached leaves) or the environmental conditions (light, nutrient availability), the symptoms of senescence differ. The aim of this work was to elucidate the catabolism of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco, EC. 4.1.1.39) under various senescence-inducing conditions. Leaf senescence was initiated in intact plants by darkness or by N-deprivation and in leaf segments by exposure to light or darkness. Depending on the treatment, a 50 kDa fragment of Rubisco was observed. The formation of this fragment was enhanced by leaf detachment and low light. In segments exposed to high light and in intact plants induced to senesce by N-deprivation, the fragment was essentially absent. Since an antibody against the N-terminus of a large subunit of Rubisco (LSU) did not cross-react with the fragment, it appears likely that a smaller fragment was removed from the N-terminus of LSU. Inhibitor studies suggest that a cysteine endopeptidase was involved in the formation of the 50 kDa fragment. Non-denaturing-PAGE followed by SDS-PAGE revealed that the fragment was produced while LSU was integrated in the holoenzyme complex, and that it remained there after being produced. It remains open how the putative endopeptidase reaches the stromal protein Rubisco. The results indicate that depending on the senescence-inducing conditions, different proteolytic enzymes may be involved. The involvement of vacuolar proteases must be considered as occurring during LSU degradation, which takes place in darkness, low light or under carbon limitation.

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Abbreviations

E-64:

trans-epoxysuccinyl-l-leucylamido-(4-guanidino)butane

GO:

glycolate oxidase

GOGAT:

glutamate synthase

LHCII:

light-harvesting chlorophyll a,b binding complex

LSU:

large subunit of Rubisco

N-LSU:

N-terminal part of LSU

PAR:

photosynthetically active radiation

PEPC:

phosphoenolpyruvate carboxylase

SSU:

small subunit of Rubisco

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Acknowledgments

We thank S.J. Crafts-Brandner, S. Gepstein, R. Houtz, T. Sugiyama, and R.M. Wallsgrove for their kind gifts of the primary antibodies used for the immunological detection of proteins and S.J. Crafts-Brandner for improving the English of the manuscript. This work was supported by the Swiss National Science Foundation (Project 31-55289.98).

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Authors and Affiliations

  1. Institute of Plant Sciences, University of Bern, Altenbergrain 21, 3013, Bern, Switzerland

    Melanie Thoenen, Beat Herrmann & Urs Feller

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  1. Melanie Thoenen
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  2. Beat Herrmann
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  3. Urs Feller
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Correspondence to Urs Feller.

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Communicated by K. Strzalka.

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Thoenen, M., Herrmann, B. & Feller, U. Senescence in wheat leaves: is a cysteine endopeptidase involved in the degradation of the large subunit of Rubisco?. Acta Physiol Plant 29, 339–350 (2007). https://doi.org/10.1007/s11738-007-0043-4

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