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The small subunit ADP-glucose pyrophosphorylase (ApS) promoter mediates okadaic acid-sensitive uidA expression in starch-synthesizing tissues and cells in Arabidopsis

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Abstract

Transgenic plants of Arabidopsis thaliana Heynh., transformed with a bacterial β-glucuronidase (GUS) gene under the control of the promoter of the small subunit (ApS) of ADP-glucose pyrophosphorylase (AGPase), exhibited GUS staining in leaves (including stomata), stems, roots and flowers. Cross-sections of stems revealed GUS staining in protoxylem parenchyma, primary phloem and cortex. In young roots, the staining was found in the root tips, including the root cap, and in vascular tissue, while the older root–hypocotyl axis showed prominent staining in the secondary phloem and paratracheary parenchyma of secondary xylem. The GUS staining co-localized with ApS protein, as found by tissue printing using antibodies against ApS. Starch was found only in cell and tissue types exhibiting GUS staining and ApS labelling, but not in all of them. For example, starch was lacking in the xylem parenchyma and secondary phloem of the root–hypocotyl axis. Sucrose potently activated ApS gene expression in leaves of wild-type (wt) plants, and in transgenic seedlings grown on sucrose medium where GUS activity was quantified with 4-methylumbelliferyl-β-glucuronide as substrate. Okadaic acid, an inhibitor of protein phosphatases 1 and 2A, completely blocked expression of ApS in mature leaves of wt plants and prevented GUS staining in root tips and flowers of the transgenic plants, suggesting a similar signal transduction mechanism for ApS expression in various tissues. The data support the key role of AGPase in starch synthesis, but they also underlie the ubiquitous importance of the ApS gene for AGPase function in all organs/tissues of Arabidopsis.

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Fig. 1a–f.
Fig. 2a–i.
Fig. 3a, b.
Fig. 4.
Fig. 5a–g.

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Abbreviations

AGPase:

ADP-glucose pyrophosphorylase

ApL:

large subunit of AGPase

ApS:

small subunit of AGPase

GUS:

β-glucuronidase

MUG:

4-methylumbelliferyl-β-glucuronide

OKA:

okadaic acid

Pi :

inorganic phosphate

PGA:

3-phosphoglyceric acid

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Acknowledgements

This work was supported, in part, by grants from the Swedish Natural Science Research Council, the Swedish Foundation For Strategic Research, and the Umeå biotechnology fund (to L.A.K.), as well as from the Foundation for Polish Science (supporting a postdoctoral scholarship for A.S.) and grant no. 6 P04C 01920 from the State Committee for Scientific Research (KBN, Poland, to I.C.).

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Authors and Affiliations

  1. Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, 901 87, Umeå, Sweden

    Françoise Martz & Leszek A. Kleczkowski

  2. Department of Plant Physiology, Maria Curie-Sklodowska University, 20-033, Lublin, Poland

    Anna Siedlecka

  3. Institute of Biology, University of Bialystok, 15-950, Bialystok, Poland

    Iwona Ciereszko

  4. Department of Forest Genetics and Plant Physiology, SLU, 901 83 , Umeå, Sweden

    Ewa Mellerowicz

  5. Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan

    Jychian Chen

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  1. Anna Siedlecka
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Correspondence to Leszek A. Kleczkowski.

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Siedlecka, A., Ciereszko, I., Mellerowicz, E. et al. The small subunit ADP-glucose pyrophosphorylase (ApS) promoter mediates okadaic acid-sensitive uidA expression in starch-synthesizing tissues and cells in Arabidopsis . Planta 217, 184–192 (2003). https://doi.org/10.1007/s00425-003-0982-y

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