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Sequence of the structural gene for granule-bound starch synthase of potato (Solarium tuberosum L.) and evidence for a single point deletion in the amf allele

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Summary

The genomic sequence of the potato gene for starch granule-bound starch synthase (GBSS; “waxy protein”) has been determined for the wild-type allele of a monoploid genotype from which an amylose-free (amf) mutant was derived, and for the mutant part of the amf allele. Comparison of the wild-type sequence with a cDNA sequence from the literature and a newly isolated cDNA revealed the presence of 13 introns, the first of which is located in the untranslated leader. The promoter contains a G-box-like sequence. The deduced amino acid sequence of the precursor of GBSS shows a high degree of identity with monocot waxy protein sequences in the region corresponding to the mature form of the enzyme. The transit peptide of 77 amino acids, required for routing of the precursor to the plastids, shows much less identity with the transit peptides of the other waxy preproteins, but resembles the hydropathic distributions of these peptides. Alignment of the amino acid sequences of the four mature starch synthases with the Escherichia coli glgA gene product revealed the presence of at least three conserved boxes; there is no homology with previously proposed starch binding domains of other enzymes involved in starch metabolism. We report the use of chimeric constructs with wild-type and amf sequences to localize, via complementation experiments, the region of the amf allele in which the mutation resides. Direct sequencing of polymerase chain reaction products confirmed that the amf mutation is a deletion of a single AT basepair in the region coding for the transit peptide. Premature termination of translation as a result of this frameshift mutation results in a small peptide. However, a protein reacting with anti-GBSS serum, slightly larger than the wild-type mature GBSS, can be detected in a membrane fraction from amylose-free tubers. A possible explanation for this phenomenon will be discussed.

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Authors and Affiliations

  1. Department of Genetics, University of Groningen, Kerklaan 30, NL-9751 NN, Haren, The Netherlands

    Feike R. van der Leij & Will J. Feenstra

  2. Department of Plant Breeding, University of Wageningen, P.O. Box 386, NL-6700 AJ, Wageningen, The Netherlands

    Richard G. F. Visser & Evert Jacobsen

  3. Biochemistry Department, University of Groningen, Nijenborgh 16, NL-9747 AG, Groningen, The Netherlands

    Anne S. Ponstein

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  1. Feike R. van der Leij
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  2. Richard G. F. Visser
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  3. Anne S. Ponstein
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  4. Evert Jacobsen
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  5. Will J. Feenstra
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Communicated by H. Saedler

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van der Leij, F.R., Visser, R.G.F., Ponstein, A.S. et al. Sequence of the structural gene for granule-bound starch synthase of potato (Solarium tuberosum L.) and evidence for a single point deletion in the amf allele. Molec. Gen. Genet. 228, 240–248 (1991). https://doi.org/10.1007/BF00282472

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  • DOI: https://doi.org/10.1007/BF00282472

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