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Effect of water stress on lupin stem protein analysed by two-dimensional gel electrophoresis

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Abstract

Lupinus albus plants can withstand severe drought stress and show signs of recovery 24 h after rewatering (RW). Two-dimensional gel electrophoresis was used to evaluate the effect of water deficit (WD) on the protein composition of the two components of the lupin stem (stele and cortex). This was performed at three distinct stress levels: an early stage, a severe WD, and early recovery. Protein characterisation was performed through mass spectrometric partial sequencing. Modifications in the protein expression were first noticed at 3 days of withholding water, when the plant water status was still unaffected but some decrease in the relative soil water content had already occurred. An increase in serine proteases, possibly associated with WD sensing, was an early alteration induced by WD. When the stress severity increased, a larger number of stem proteins were affected. Immunophilin, serine protease and cysteine protease (well-known components of animal sensing pathways) were some of these proteins. The simultaneous expression of proteases and protease inhibitors that reacted differently to the stress level and to RW was found. Although the level of protease inhibitors was significantly raised, RW did not cause de novo expression of proteins. Many amino acid sequences did not match known sequences of either protein or expressed sequence tag databases. This emphasises the largely unknown nature of stem proteins. Nevertheless, some important clues regarding the way the lupin plant copes with WD were revealed.

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Abbreviations

DAW:

Days of withholding water

MS:

Mass spectrometry

RWC:

Relative water content

RSWC:

Relative soil water content

2DE:

Two-dimensional electrophoresis

RW:

Rewatering

WD:

Water deficit

Ψ pd:

Leaf water potential at pre-dawn

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Acknowledgements

We thank Profs. C. Arruda Pacheco and R. Pinto Ricardo (ISA) for their help in soil characterisation and Prof. M. Manuela Chaves for helpful discussions. C. Pinheiro acknowledges a grant from FCT (PRAXIS BD/16137/98).

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Authors and Affiliations

  1. Instituto de Tecnologia Química e Biológica, Apartado127, 2781901, Oeiras, Portugal

    C. Pinheiro & C. P. Ricardo

  2. Max-Planck-Institut für Molekulare Pflanzenphysiologie, 14424, Potsdam, Germany

    J. Kehr

  3. Instituto Superior de Agronomia, Tapada da Ajuda, 1349017, Lisbon, Portugal

    C. P. Ricardo

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  1. C. Pinheiro
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  2. J. Kehr
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  3. C. P. Ricardo
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Correspondence to C. P. Ricardo.

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Pinheiro, C., Kehr, J. & Ricardo, C.P. Effect of water stress on lupin stem protein analysed by two-dimensional gel electrophoresis. Planta 221, 716–728 (2005). https://doi.org/10.1007/s00425-004-1478-0

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