Photosynthesis Research

, Volume 18, Issue 1–2, pp 33–59 | Cite as

Transposon mutagenesis of nuclear photosynthetic genes in Zea mays

  • William B. Cook
  • Donald Miles
Genes Minireview


The discovery of a new maize (Zea mays L.) transposon system, Mutator, and the cloning of the 1.4 kilobase transposon, Mul, have made feasible the isolation of nuclear photosynthetic genes which are recognized only by their mutant phenotype. Mutant maize plants which express a high chlorophyll fluorescent (hcf) phenotype due to a defect in the electron transport or photophosphorylation apparatus have been isolated following mutagenesis with an active Mutator stock. The affected genes and their products in these mutants are inaccessible to classical methods of analysis. However, mutagenesis with the Mutator transposon makes it possible to isolate these genes.

Although the PSII-deficient mutant hcf3 has been thoroughly studied by classical photo-biological methods, the nature of the lesion which results in the observed phenotype has not been established. A Mutator-induced allele of hcf3 has been isolated. A fragment of genomic DNA has been identified which is homologous to Mul and co-segregates with the mutant phenotype. This fragment is expected to contain a portion of the hcf3 locus which will be used to clone the normal gene. Direct study of the gene can provide insight into the nature and function of its polypeptide product.

This approach can be used to study any photosynthetic gene which has been interrupted by a transposon. The isolation of more than 100 different chemically-induced hcf mutants, most of which can not be fully characterized using classical means, indicates the wealth of information which can be obtained using a transposon tagging technique.

Key words

cloning mutation photosynthesis mutant transposon 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alleman M and Freeding M (1986) The Mu transposable elements of maize: Evidence for transposition and copy number regulation during development. Genetics 112: 107–119PubMedGoogle Scholar
  2. Baker B, Schell J, Lorz H and Federoff NV (1986) Transposition of the maize controlling element ‘Activator’ in tobacco. Proc Natl Acad Sci USA 83: 4844–4848Google Scholar
  3. Barkan A, Miles D and Taylor WC (1986) Chloroplast gene expression in nuclear, photosynthetic mutants in maize. EMBO J 5: 1421–1427PubMedGoogle Scholar
  4. Barker RF, Thompson DV, Talbot DR, Swanson J and Bennetzen JL (1984) Nucleotide sequence of the maize transposable element Mul. Nucleic Acids Res 12: 5955–5967PubMedGoogle Scholar
  5. Beckett JB (1978) B-A translocation in maize. I. Use in locating genes by chromosome arms. J Heredity 69: 27–36.Google Scholar
  6. Bennetzen JL, Swanson J, Taylor WC and Freeling WC (1984) DNA insertion in the first intron of maize Adhl affects message levels: Cloning of progenitor and mutant alleles. Proc Natl Acad Sci USA 81: 4125–4128PubMedGoogle Scholar
  7. Bennetzen JL (1987) Covalent DNA modification and the regulation of Mutator element transposition in maize. Mol Gen Genet 208: 45–51CrossRefGoogle Scholar
  8. Bennetzen JL, Fracasso RP, Morris DW, Robertson DS and Skogen-Hagenson MJ (1987) Concomitant regulation of Mu1 transposition and Mutator activity in maize. Mol Gen Genet 208: 57–62CrossRefGoogle Scholar
  9. Bonas U, Sommer H, Harrison BJ and Saedler H (1984) The transposable element Taml of Antirrhinum majus is 17 kb long. Mol Gen Genet 194: 138–143Google Scholar
  10. Calos MP and Miller JH (1980) Transposable elements. Cell 20: 579–595CrossRefPubMedGoogle Scholar
  11. Campbell A (1981) Evolutionary significance of accessory DNA elements in bacteria. Ann Rev Microbiol 35: 55–83CrossRefGoogle Scholar
  12. Carpenter R, Martin C and Coen ES (1987) Comparison of genetic behavior of the transposable element Tam3 at two unliked pigment loci in Antirrhinum majus. Mol Gen Genet 207: 82–89CrossRefGoogle Scholar
  13. Chandler VL and Walbot V (1986) DNA modification of a maize transposable element correlates with loss of activity. Proc Natl Acad Sci USA 83: 1767–1771PubMedGoogle Scholar
  14. Chandler VL, Rivin C and Walbot V (1986) Stable non-Mutator stocks of maize have sequences homologous to the Mu1 transposable element. Genetics 114: 1007–1021PubMedGoogle Scholar
  15. Chen J (1986) Plant genomic DNA preparation. In: Plant Gene Cloning Manual. Cold Spring Harbor: Maize molecular genetics group, Cold Spring Harbor LaboratoryGoogle Scholar
  16. Cone KC, Burr FA and Burr B (1986) Molecular analysis of the maize anthocyanin regulatory locus C1. Proc Natl Acad Sci USA 83: 9631–9635PubMedGoogle Scholar
  17. Cook B and Miles D (1987) Mutator-induced PSII photosynthesis mutant is allelic to hcf3. Maize Genet Coop Newslett 61: 44Google Scholar
  18. Cook B, Hunt M and Miles D (1987) Mutator-induced mutation on 8L affects the chloroplast cytochrome b6/f complex. Maize Genet Coop Newslett 61: 44Google Scholar
  19. Federoff NV (1983) Controlling elements in maize. In: Shapiro JA (ed) Mobile Genetic Elements, pp 1–65. New York: Academic Press/Harcourt Brace JovanovichGoogle Scholar
  20. Federoff NV, Wessler S and Shure M (1983) Isolation of the transposable maize controlling elements Ac and Ds, Cell 35: 235–242CrossRefPubMedGoogle Scholar
  21. Federoff NV, Furtek DB and Nelson OE (1984) Cloning of the bronze locus in maize by a simple and generalizable procedure using the transposable controlling element Activator (Ac). Proc Natl Acad Sci USA 81: 3825–3829Google Scholar
  22. Feinberg AP and Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Bioch 132: 6–13 and Addendum Anal Bioch 137: 266–267PubMedGoogle Scholar
  23. Freeling M (1984) Plant transposable elements and insertion sequences. Ann Rev Plant Physiol 35: 277–298Google Scholar
  24. Goldberg RB, Hoschek G and Vodkin LO (1983) An insertion sequence blocks the expression of a soybean lectin gene. Cell 33: 465–475CrossRefPubMedGoogle Scholar
  25. Grindley NDF and Reed RR (1985) Transpositional recombination in prokaryotes. Ann Rev Biochem 54: 863–896CrossRefPubMedGoogle Scholar
  26. Guikema JA and Sherman LA (1980) Electrophoretic profiles of cyanobacterial membrane polypeptides showing heme-dependent peroxidase activity. Biochem Biophys Acta 637: 189–201Google Scholar
  27. Heffron F (1983) Tn3 and its relatives. In: Shapiro JA (ed) Mobile Genetic Elements, pp 223–260. New York: Academic Press/Harcourt Brace JovanovichGoogle Scholar
  28. Hehl R, Sommer H and Saedler H (1987) Interaction between Tam1 and Tam2 transposable elements of Antirrhinum majus. Mol Gen Genet 207: 47–53CrossRefGoogle Scholar
  29. Helentjaris T, King G, Slocum M, Siedenstrang C and Wegman S (1985) Restriction fragment polymorphisms as probes for plant diversity and their development as tools for applied plant breeding. Plant Mol Biol 5: 109–118Google Scholar
  30. Hudson A, Carpenter R and Coen ES (1987) De novo activation of the transposable element Tam2 of Antirrhinum majus. Mol Gen Genet 207: 54–59CrossRefGoogle Scholar
  31. Iida S, Meyer J and Arber W (1983) Prokaryotic IS elements. In: Shapiro JA (ed) Mobile Genetic Elements, pp 159–222. New York: Academic Press/Harcourt Brace JovanovichGoogle Scholar
  32. Kleckner N (1981) Transposable elements in prokaryotes. Ann Rev Genet 15: 341–404CrossRefPubMedGoogle Scholar
  33. Krueger RW and Miles D (1981) Photosynthesis in tall fescue. I. High rates of electron transport and photophosphorylation in chloroplasts of hexaploid plants. Plant Physiol 67: 763–767Google Scholar
  34. Leto KJ and Miles D (1980) Characterization of three Photosystem II mutants in Zea mays L. lacking a 32,000 Dalton lamellar polypeptide. Plant Physiol 66: 18–24Google Scholar
  35. Lillis M and Freeling M (1986) Mu transposons in maize. Trends Genet 2: 183–188CrossRefGoogle Scholar
  36. Martienssen RA, Barkan A, Scriven A and Taylor WC (1987) Identification of a nuclear gene involved in thylakoid structure. In: Leaver C and Sze H (Eds) Plant Membranes: Structures, Function and biogenesis (UCLA Symposia on Molecular and Cellular Biology 63: 118–194). New York: Alan R LissGoogle Scholar
  37. McClintock B (1984) the significance of responses of the genome to challenge. Science 226: 792–801Google Scholar
  38. Metz JG and Miles D (1982) Use of a nuclear mutant of maize to identify components of photosystem II. Biochem Biophys Acta 681: 95–102Google Scholar
  39. Metz JG, Miles D and Rutherford AW (1983) Characterization of nuclear mutants of maize which lack the cytochrome f/b-563 complex. Plant Physiol 73: 452–459Google Scholar
  40. Miles D (1980) Mutants of higher plants: Maize. In: San Pietro A (ed) Photosynthesis, Part C, Methods in Enzymology 69: 3–22. New York: Academic PressGoogle Scholar
  41. Miles D (1982) the use of mutations to probe photosynthesis in higher plants. In: Edelman M, Hallick RB and Chua N-H (eds) Methods in Chloroplast Molecular Biology, pp 75–107. Amsterdam: Elsevier Biomedical PressGoogle Scholar
  42. Miles D and Randall D (1983) Nuclear mutants of maize altering the large subunit of ribulose-1,5-bisphosphate carboxylase. In: Randall DD, Blevins DG and Larson R (eds) Current Topics in Plant Biochemistry and Physiology 1: 231. Columbia: Interdisciplinary Plant Biochemistry and Physiology ProgramGoogle Scholar
  43. Miles D, Leto KJ, Neuffer MG, Polacco M, Hanks JF and Hunt MA (1985) Chromosome arm location of photosynthesis mutants in Zea mays L. using B-A translocations. In: Steinbeck KE, Arntzen CJ and Bogorad J (Eds) Molecular biology of the photosynthetic apparatus, pp 361–365. Cold spring Harbor: Cold Spring Harbor LaboratoryGoogle Scholar
  44. Nevers P, Shepherd NS and Saedler H (1986) Plant transposable elements. Adv Bot Res 12: 103–203Google Scholar
  45. O'Reilly C, Shepherd NS, Pereira A, Schwarz-Sommer Zs, Bertram I, Robertson DS, Peterson PA and Saedler H. (1985) Molecular cloning of the a1 locus of Zea mays using the transposable elements En and Mu1. EMBO J 4: 877–882Google Scholar
  46. Paz-Ares J, Wienand U, Peterson PA and Saedler H (1986) Molecular cloning of the c locus of Zea mays: a locus regulating the anthocyanin pathway. EMBO J 5: 829–833Google Scholar
  47. Pereira A, Schwarz-Sommer Zs, Gierl A, Bertram I, Peterson PA and Saedler H (1985) Genetic and molecular analysis of the Enhancer (En) transposable element system of Zea maus. EMBO J 4: 17–23Google Scholar
  48. Pohlman RF, Federoff NV and Mesing J (1984) the nucleotide sequence of the maize controlling element Activator. Cell 37: 635–643CrossRefPubMedGoogle Scholar
  49. Robertson GS (1978) Characterization of a mutator system in maize. Mutation Res 51: 21–28Google Scholar
  50. Roeder GS and Fink GR (1983) Transposable elements in yeast. In: Shapiro JA (ed) Mobile Genetic Elements, pp 300–328. New York: Academic Press/Harcourt Brace JovanovichGoogle Scholar
  51. Rubin GM (1983) Dispersed repetitive DNAs in Drosophila. In: Shapiro JA (ed) Mobile Genetic Elements, pp 329–362. New York: Academic Press/Harcourt Brace JovanovichGoogle Scholar
  52. Sachs MM, Peacock WJ, Dennis ES and Gerlack WL (1983) Maize Ac/Ds controlling elements. A molecular viewpoint. Maydica 28: 289–303Google Scholar
  53. Schwartz D and Dennis E (1986) Transposase activity of the Ac controlling element in maize is regulated by its degree of methylation. Mol Gen Genet 206: 476–482CrossRefGoogle Scholar
  54. Schwarz-Sommer Zs, Gierl A, Klosgen RB, Wienand U, Peterson PA and Saedler H (1984) The Spm (En) transposable element controls the excision of a 2-kb DNA insert at the wxm−8 allele of Zea mays. EMBO J 3: 1021–1028Google Scholar
  55. Shapiro JA (1983) Genetic reorganization in cell lineages. In: Shapiro JA, (ed) Mobile Genetic Elements, pp xi-xvi. New York: Academic Press/Harcourt Brace JovanovichGoogle Scholar
  56. Shure M, Wessler S and Federoff N (1983) Molecular identification and isolation of the Waxy locus in maize. Cell 35: 225–233PubMedGoogle Scholar
  57. Somerville CR (1986) Analysis of photosynthesis with mutants of higher plants and algae. Ann Rev Plant Physiol 37: 467–507Google Scholar
  58. Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517PubMedGoogle Scholar
  59. Taylor LP, Chandler VL and Walbot V (1986) Insertion of 1.4 kb and 1.7 kb Mu elements into the Bronze-1 gene of Zea mays L. Maydica 31: 31–45Google Scholar
  60. Taylor WC, Barkan A and Martienssen RA (1988) The use of nuclear mutants in the analysis of chloroplast development. Developmental Genet 8: 305–320Google Scholar
  61. Thomas PE, Rayn D and Levin W (1976) An improved staining procedure for the detection of the peroxidase activity of cytochrome P-450 on sodium dodecyl sulfate polyacrylamide gels. Anal Biochem 75: 168–176PubMedGoogle Scholar
  62. Tittgen J, Hermans J, Steppuhn J, Jansen T, Jannson C, Anderson B, Nechushtai R, Nelson N and Herrmann RG (1986) Isolation of cDNA clones for fourteen nuclear-encoded thylakoid membrane proteins. Mol Gen Genet 204: 258–265Google Scholar
  63. Walbot V, Chandler VL, Taylor LP and McLaughlin P (1987) Regulation of transposable element activities during the development and evolution of Zea mays L. In: Development as an Evolutionary Process, pp 265–284. New York: Alan R LissGoogle Scholar
  64. Webber DF (1986) The production and utilisation of monosomic Zea mays in cytogenetic studies. In: Reddy GM and Co EH (eds) Gene Structure and Function in Higher Plants, pp 191–204. Dehli: Oxford and IBH PublishingGoogle Scholar
  65. Young RA and Davis RW (1983) Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci USA 80: 1194–1198PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1988

Authors and Affiliations

  • William B. Cook
    • 1
  • Donald Miles
    • 1
  1. 1.Department of Biological SciencesUniversity of MissouriColumbiaUSA

Personalised recommendations