Skip to main content
NCBI home page
Plant Physiology logoLink to Plant Physiology
. 1988 Apr;86(4):1131–1135. doi: 10.1104/pp.86.4.1131

Isolation and Characterization of a Starchless Mutant of Arabidopsis thaliana (L.) Heynh Lacking ADPglucose Pyrophosphorylase Activity 1

Tsan-Piao Lin 1,2,2, Timothy Caspar 1,2, Chris Somerville 1,2, Jack Preiss 1,2
PMCID: PMC1054640  PMID: 16666044

Abstract

A mutant of Arabidopsis thaliana lacking ADPglucose pyrophosphorylase activity (EC 2.7.7.27) was isolated (from a mutagenized population of plants) by screening for the absence of leaf starch. The mutant grows as vigorously as the wild type in continuous light but more slowly than the wild type in a 12 hours light/12 hours dark photoperiod. Genetic analysis showed that the deficiency of both starch and ADPglucose pyrophosphorylase activity were attributable to a single, nuclear, recessive mutation at a locus designated adg1. The absence of starch in the mutant demonstrates that starch synthesis in the chloroplast is entirely dependent on a pathway involving ADPglucose pyrophosphorylase. Analysis of leaf extracts by two-dimensional polyacrylamide gel electrophoresis followed by Western blotting experiments using antibodies specific for spinach ADPglucose pyrophosphorylase showed that two proteins, present in the wild type, were absent from the mutant. The heterozygous F1 progeny of a cross between the mutant and wild type had a specific activity of ADPglucose pyrophosphorylase indistinguishable from the wild type. These observations suggest that the mutation in the adg1 gene in TL25 might affect a regulatory locus.

Full text

PDF
1131

Images in this article

1134Fig. 3
on p.1134

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  2. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  3. Dickinson D. B., Preiss J. Presence of ADP-Glucose Pyrophosphorylase in Shrunken-2 and Brittle-2 Mutants of Maize Endosperm. Plant Physiol. 1969 Jul;44(7):1058–1062. doi: 10.1104/pp.44.7.1058. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hannah L. C., Nelson O. E. Characterization of adenosine diphosphate glucose pyrophosphorylases from developing maize seeds. Plant Physiol. 1975 Feb;55(2):297–302. doi: 10.1104/pp.55.2.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hawker J. S., Ozbun J. L., Ozaki H., Greenberg E., Preiss J. Interaction of spinach leaf adenosine diphosphate glucose alpha-1,4-glucan alpha-4-glucosyl transferase and alpha-1,4-glucan, alpha-1,4-glucan-6-glycosyl transferase in synthesis of branched alpha-glucan. Arch Biochem Biophys. 1974 Feb;160(2):530–551. doi: 10.1016/0003-9861(74)90430-5. [DOI] [PubMed] [Google Scholar]
  6. Hawkes R. Identification of concanavalin A-binding proteins after sodium dodecyl sulfate--gel electrophoresis and protein blotting. Anal Biochem. 1982 Jun;123(1):143–146. doi: 10.1016/0003-2697(82)90634-0. [DOI] [PubMed] [Google Scholar]
  7. Jones M. G., Outlaw W. H., Lowry O. H. Enzymic assay of 10 to 10 moles of sucrose in plant tissues. Plant Physiol. 1977 Sep;60(3):379–383. doi: 10.1104/pp.60.3.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jones T. W., Gottlieb L. D., Pichersky E. Reduced enzyme activity and starch level in an induced mutant of chloroplast phosphoglucose isomerase. Plant Physiol. 1986 Jun;81(2):367–371. doi: 10.1104/pp.81.2.367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Morell M. K., Bloom M., Knowles V., Preiss J. Subunit Structure of Spinach Leaf ADPglucose Pyrophosphorylase. Plant Physiol. 1987 Sep;85(1):182–187. doi: 10.1104/pp.85.1.182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Okita T. W., Greenberg E., Kuhn D. N., Preiss J. Subcellular localization of the starch degradative and biosynthetic enzymes of spinach leaves. Plant Physiol. 1979 Aug;64(2):187–192. doi: 10.1104/pp.64.2.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Preiss J., Okita T. W., Greenberg E. Characterization of the spinach leaf phosphorylases. Plant Physiol. 1980 Nov;66(5):864–869. doi: 10.1104/pp.66.5.864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Preiss J., Sabraw A., Greenberg E. An ADP-glucsoe pyrophosphorylase with lower apparent affinities for substract and effector molecules in an Escherichia coli B mutant deficient in glycogen synthesis. Biochem Biophys Res Commun. 1971 Jan 22;42(2):180–186. doi: 10.1016/0006-291x(71)90085-4. [DOI] [PubMed] [Google Scholar]
  14. Rufty T. W., Huber S. C. Changes in Starch Formation and Activities of Sucrose Phosphate Synthase and Cytoplasmic Fructose-1,6-bisphosphatase in Response to Source-Sink Alterations. Plant Physiol. 1983 Jun;72(2):474–480. doi: 10.1104/pp.72.2.474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  16. Tsai C. Y., Nelson O. E. Starch-deficient maize mutant lacking adenosine dephosphate glucose pyrophosphorylase activity. Science. 1966 Jan 21;151(3708):341–343. doi: 10.1126/science.151.3708.341. [DOI] [PubMed] [Google Scholar]
  17. Wallsgrove R. M., Turner J. C., Hall N. P., Kendall A. C., Bright S. W. Barley mutants lacking chloroplast glutamine synthetase-biochemical and genetic analysis. Plant Physiol. 1987 Jan;83(1):155–158. doi: 10.1104/pp.83.1.155. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES