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Purification and partial characterisation of two abscisic-acid-responsive proteins induced in cultured embryos ofPisum sativum L.

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Abstract

When pea (Pisum sativum L.) embryos were cultured on low osmotica, with or without added abscisic acid (ABA), there was very little change in the total mRNA translation products resolved by one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The only marked alteration was an increase in production of two low-molecular-weight proteins. The purification and partial characterisation of these two ABA-responsive seed proteins (ABR17 and ABR18) is described. Both proteins were purified to homoeneity, as judged by SDS-PAGE, from embryos cultured in the presence of ABA. Antisera were raised against both proteins. Each serum cross-reacted with the other protein, indicating that the proteins are closely related. Their apparent molecular masses (Mrs) were estimated to be 17200 (ABR17) and 18100 (ABR18) by SDS-PAGE, and 26000 by gel filtration. Both proteins were heterogeneous on isoelectric focusing. Neither protein was detected (by immunoblotting or immunoprecipitation of cell-free translation products) in embryos grown in vivo at early to mid-development stages but both were present in embryos late in development. These proteins appear to be produced late in seed development but are capable of being induced early in development by culturing embryos in vitro and are markedly enhanced by ABA.

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Abbreviations

ABA:

(±)cis, trans-abscisic acid

ABR17:

Mr-17200 ABA-responsive protein

ABR18:

Mr-18100 ABA-responsive protein

FW:

fresh weight

IgG:

immuno-globulin G

Mr :

apparent molecular mass

pI:

isoelectric point

SDS-PAGE:

sodium dodecyl sulphate-polyacrylamide gel electrophoresis

Tricine:

N-[2-hydroxy-1,1-bis(hydroxymethy)ethyl]glycine

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Authors and Affiliations

  1. Department of Agricultural Sciences, University of Bristol, AFRC Institute of Arable Crops Research, Long Ashton Research Station, BS18 9AF, Bristol, UK

    D. H. P. Barratt

  2. John Innes Institute and AFRC Institute of Plant Science Research, Colney Lane, NR4 7UH, Norwich, UK

    C. Domoney & T. L. Wang

Authors
  1. D. H. P. Barratt
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  2. C. Domoney
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  3. T. L. Wang
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Barratt, D.H.P., Domoney, C. & Wang, T.L. Purification and partial characterisation of two abscisic-acid-responsive proteins induced in cultured embryos ofPisum sativum L.. Planta 180, 16–23 (1989). https://doi.org/10.1007/BF02411405

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  • DOI: https://doi.org/10.1007/BF02411405

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