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Ecotype dependent expression and alternative splicing of epithiospecifier protein (ESP) in Arabidopsis thaliana

Plant Molecular Biology volume 78, pages 361–375 (2012)Cite this article

Abstract

Epithiospecifier protein (ESP) is responsible for diverting glucosinolate hydrolysis from the generation of isothiocyanates to that of epithionitriles or nitriles, and thereby negatively affects the ability of the plant to defend itself against certain insects. Despite this important role of ESP, little is known about its expression in plant tissues and the regulation thereof. We therefore investigated ESP expression by qPCR and Western blot in different organs during the growth cycle of the two Arabidopsis thaliana ecotypes Col-0 and Mt-0. Besides the fact that ESP transcript and protein levels were revealed to be much higher in Mt-0 than in Col-0 in all cases, our qPCR results also indicated that ESP expression is regulated differently in the two A. thaliana ecotypes. No ESP protein was detected by Western blot in any organ or developmental stage for Col-0. During the assays an alternative splice variant of ESP was identified in Col-0, but not Mt-0, leading to a mis-spliced transcript which could explain the low expression levels of ESP in the former ecotype. Analysis of genomic sequences containing the ESP splice sites, of ESP protein level and ESP activity from seven A. thaliana ecotypes showed a positive correlation between the presence of a non-canonical 5′ splice site for ESP and the absence of detectable ESP protein levels and ESP activity. When analysing the expression of both transcript variants in Col-0 after treatment with methyl jasmonate, a condition known to “induce ESP”, it was indeed the alternative splice variant that was preferentially induced.

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Acknowledgments

We thank Bente U. Halvorsen and Torfinn Sparstad for excellent technical assistance. The work was supported by the Norwegian Research Council and the Nordic Joint Committee for Agricultural Research through grants 175691/NKJ126, 185173/V40 and 184146/S10.

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Affiliations

  1. Department of Biology, Norwegian University of Science and Technology (NTNU), Realfagbygget, Høgskoleringen 5, 7491, Trondheim, Norway

    R. Kissen, E. Hyldbakk, C.-W. V. Wang, C. G. Sørmo & A. M. Bones

  2. Division of Biology, Imperial College London, Sir Alexander Fleming Building, Exhibition Road, South Kensington, London, SW7 2AZ, UK

    J. T. Rossiter

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  1. R. Kissen
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  2. E. Hyldbakk
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  3. C.-W. V. Wang
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  4. C. G. Sørmo
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  5. J. T. Rossiter
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  6. A. M. Bones
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Correspondence to A. M. Bones.

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Kissen, R., Hyldbakk, E., Wang, CW.V. et al. Ecotype dependent expression and alternative splicing of epithiospecifier protein (ESP) in Arabidopsis thaliana . Plant Mol Biol 78, 361–375 (2012). https://doi.org/10.1007/s11103-011-9869-7

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  • DOI: https://doi.org/10.1007/s11103-011-9869-7

Keywords

  • Glucosinolates
  • Epithiospecifier protein
  • Arabidopsis thaliana
  • Ecotypes
  • Alternative splice variant
  • qPCR