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Protein changes and proteolytic degradation in red and white clover plants subjected to waterlogging

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Abstract

Red (Trifolium pratense L., cv. “Start”) and white clover varieties (Trifolium repens L., cv. “Debut” and cv. “Haifa”) were waterlogged for 14 days and subsequently recovered for the period of 21 days. Physiological and biochemical responses of the clover varieties were distinctive, which suggested different sensitivity toward flooding. The comparative study of morphological and biochemical parameters such as stem length, leaflet area, dry weight, protein content, protein pattern and proteolytic degradation revealed prominent changes under waterlogging conditions. Protease activity in the stressed plants increased significantly, especially in red clover cv. “Start”, which exhibited eightfold higher azocaseinolytic activity compared to the control. Changes in the protein profiles were detected by SDS-PAGE electrophoresis. The specific response of some proteins (Rubisco, Rubisco-binding protein, Rubisco activase, ClpA and ClpP protease subunits) toward the applied stress was assessed by immunoblotting. The results characterized the red clover cultivar “Start” as the most sensitive toward waterlogging, expressing reduced levels of Rubisco large and small subunits, high content of ClpP protease subunits and increased activity of protease isoforms.

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Abbreviations

Clp:

ATP-dependent protease belonging to serine protease

DW:

Dry weight

EDTA:

Ethylenediaminetetraacetic acid

FW:

Fresh weight

M:

Protein marker

MW:

Molecular weight

P:

Band with proteolytic activity

PHMB:

Para-chloro-mercuribenzoate

PMSF:

Phenylmethylsulfonyl fluoride

Rubisco:

Ribulose-1,5-bisphosphate carboxylase/oxygenase

RA:

Rubisco activase

RBP:

Rubisco-binding protein

RLS:

Rubisco large subunit

RSS:

Rubisco small subunit

RT:

Room temperature

RuBP:

Ribulose-1,5-bisphosphate

SDS-PAGE:

SDS polyacrylamide gel electrophoresis

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Acknowledgments

The authors thank Prof. Ts. Mihovski from the Institute of Mountain Stockbreeding and Agriculture, Troyan, Bulgaria for kindly providing cv. “Debut” seeds. This study was supported by Bulgarian National Science Foundation (project No. BG051PO001-3.3.06-0025).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Plant Stress Molecular Biology Department, Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bldg 21, 1113, Sofia, Bulgaria

    Veselin Stoychev, Lyudmila Simova-Stoilova, Irina Vaseva, Anelia Kostadinova, Rosa Nenkova & K. Demirevska

  2. Institute of Plant Sciences and Oeschger Centre for Climate Changes Research (OCCR), University of Bern, Altenbergrain 21, 3013, Bern, Switzerland

    Urs Feller

Authors
  1. Veselin Stoychev
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  2. Lyudmila Simova-Stoilova
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  3. Irina Vaseva
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  4. Anelia Kostadinova
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  5. Rosa Nenkova
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  6. Urs Feller
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  7. K. Demirevska
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Correspondence to Veselin Stoychev.

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Communicated by Z.-L. Zhang.

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Stoychev, V., Simova-Stoilova, L., Vaseva, I. et al. Protein changes and proteolytic degradation in red and white clover plants subjected to waterlogging. Acta Physiol Plant 35, 1925–1932 (2013). https://doi.org/10.1007/s11738-013-1231-z

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  • DOI: https://doi.org/10.1007/s11738-013-1231-z

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