Molecular Breeding

, Volume 10, Issue 1–2, pp 31–44 | Cite as

Stable expression of Phytase (phyA) in canola (Brassica napus) seeds: towards a commercial product

  • Anne S. Ponstein
  • Jacob B. Bade
  • Theo C. Verwoerd
  • Lucy Molendijk
  • Joost Storms
  • Rob F. Beudeker
  • Jan Pen


TheAspergillus niger gene encoding phytase(phyA) was expressed in canola (Brassicanapus). Phytase expression is controlled by the seed-specificcruciferin (CruA) promoter. Secretion of the enzyme was aimed for byincorporating the cruciferin signal peptide in the expression construct.Transgenic canola lines were generated by Agrobacteriummediated transformation using nptII as the selectable marker. Ninety-fiveindependent transgenic events were generated. Phytase expression in the T1seedsranged from 0 to 600 U/g seed. Single-copy lines were selected(based on segregation for kanamycin resistance, phytase expression and Southernanalyses) from originally multi-copy transgenic lines. Phytase was expressed inthese sub-lines up to 103 U/g. Expression levels were monitoredthrough an additional 3–4 generations (in the greenhouse and in thefield)and the accumulation of phytase appeared to be fairly stable. In the expressionrange studied, phytase expression was gene-dosage dependent.

Brassica napus Cruciferin promoter Phytase Seed-specific expression 


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Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Anne S. Ponstein
    • 1
  • Jacob B. Bade
    • 1
  • Theo C. Verwoerd
    • 1
  • Lucy Molendijk
    • 1
  • Joost Storms
    • 1
  • Rob F. Beudeker
    • 2
  • Jan Pen
    • 3
  1. 1.Syngenta MogenLeidenThe Netherlands
  2. 2.DelftThe Netherlands
  3. 3.PlantZymeLeidenThe Netherlands

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