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Analysis of the expression of potato uridinediphosphate-glucose pyrophosphorylase and its inhibition by antisense RNA

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Abstract

The expression of the enzyme UDP-glucose pyrophosphorylase (UGPase; EC 2.7.7.9) from potato (Solanum tuberosum L.) was analysed with respect to sink-source interactions and potato tuber storage. The highest level of expression was found in developing tubers, the strongest sink tissue. Storage of mature tubers at low temperatures led to an increase of the steady-state level of UGPase mRNA, implicating a role of this enzyme in the process of “cold-sweetening”. Transgenic plants were created expressing UGPase antisensee RNA under the control of the 35S promoter of the Cauliflower Mosaic Virus with the polyadenylation signal of the octopine-synthase gene. Regenerated plants were tested for reduction of UGPase at the RNA, protein and activity levels. Plants with a 95%–96% reduction of UGPase activity in growing tubers showed no change in growth and development. Also, carbohydrate metabolism in tubers of these plants was not substantially affected, indicating that only 4% of the wild-type UGPase activity is sufficient for the enzyme to function in plant growth and development.

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Abbreviations

cDNA:

copy DNA

CaMV:

Cauliflower Mosaic Virus

Glc1P:

glucose-1-phosphate

UDPGlc:

UDP-glucose

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

UGPase:

UDP-glucose pyrophosphorylase

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Author notes

  1. R. Zrenner & U. Sonnewald

    Present address: Institut für Pflanzengenetik und Kulturpflanzenforschung, Corrensstrasse 3, O-4325, Gatersleben, Germany

Authors and Affiliations

  1. Institut für Genbiologische Forschung Berlin GmbH, Ihnestrasse 63, 33, Berlin, Germany

    R. Zrenner, L. Willmitzer & U. Sonnewald

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  1. R. Zrenner
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  2. L. Willmitzer
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  3. U. Sonnewald
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Additional information

We are grateful to Dr. J.P. Spychalla (Cambridge Laboratory, Norwich, Norfolk, UK) for providing antiserum directed against the potato tuber UGPase protein. We thank J. Bergstein and B. Schäfer for photographic work, J. Dietze for plant transformation and R. Breitfeld and B. Burose for taking care of the greenhouse plants.

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Zrenner, R., Willmitzer, L. & Sonnewald, U. Analysis of the expression of potato uridinediphosphate-glucose pyrophosphorylase and its inhibition by antisense RNA. Planta 190, 247–252 (1993). https://doi.org/10.1007/BF00196618

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  • DOI: https://doi.org/10.1007/BF00196618

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