Plant Cell Reports

, Volume 19, Issue 11, pp 1041–1048 | Cite as

PIG-mediated cassava transformation using positive and negative selection

  • P. Zhang
  • J. Puonti-Kaerlas
CELL BIOLOGY AND MORPHOGENESIS

Abstract

 In order to develop new selection systems for production of transgenic cassava (Manihot esculenta Crantz), two different selection regimes were assessed for their efficiency on regeneration of transgenic cassava plants: positive selection using mannose and negative selection using hygromycin. Explants from somatic cotyledons and embryogenic suspensions were used as target tissues in the transformation experiments and bombarded using the particle inflow gun. Different culture and selection strategies were assessed to optimise the selection protocols. For the first time transgenic plants could be obtained using positive, and in the case of embryogenic suspensions, hygromycin-based negative selection. The stably transformed nature of the regenerated cassava plant lines and the expression of the transgenes were verified with PCR, RT-PCR, Southern and northern analyses. A rooting test for transgenic plants on a medium supplemented with mannose was developed to further improve the efficacy of the positive selection system. Our results demonstrate that it is possible to obtain transgenic cassava plants using non-antibiotic positive selection.

Key words Cassava (Manihot esculenta Crantz) Transformation Biolistics Positive selection Negative selection 
Abbreviations2,4-D 2,4-Dichlorophenoxyacetic acid BA 6-Benzylaminopurine CBM Basic medium CEM Elongation medium CIM Embryo induction medium CMM Maturation medium COM Organogenesis medium CTM Transformation medium GD Gresshoff and Doy medium IBA Indole-3-butyric acid FEC Friable embryogenic callus NAAα-Naphthalene acetic acid MSM Solid mannose selection medium MSN FEC Formation and embryo conversion medium SCV Settled cell volume SH Schenk and Hildebrandt medium SHM Liquid mannose selection medium 

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Copyright information

© Springer-Verlag Berlin Heidelberg 2000

Authors and Affiliations

  • P. Zhang
    • 1
  • J. Puonti-Kaerlas
    • 1
  1. 1.Institute for Plant Sciences, ETH-Zentrum / LFW E 17, CH-8092 Zürich, Switzerland e-mail: johanna.puonti-kaerlas@ipw.biol.ethz.ch Fax: +41-1-6321044CH

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