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Cyanamide hydratase as selectable marker in potato

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Abstract

The selectable marker cyanamide hydratase/cyanamide was successfully used to generate transgenic potato plants from the cultivars Russet Burbank, Ranger Russet, Bintje, Desiree, Kardal and Pentland Dell. Up to 3,000 transgenics per person per year were produced. The efficiency of transgenic production varied among cultivars, and was in general 2–3 times lower than the transformation efficiency (TE) using the selectable marker kanamycin. Differences between cultivars in sensitivity to cyanamide selection were observed, but in general a concentration of 30 mg/l was applied for selection of transgenic shoots. A stepwise increase of cyanamide concentration during the transformation procedure for Russet Burbank resulted in an improved TE via a reduction of the escape efficiency from 69 to 29 %. The cultivars differed in the hormone concentration and duration (2,4-D and Zeatin riboside) required for the production of transgenics, predominantly during the phase of shoot initiation. Only for cultivars Bintje and Pentland Dell, adaptations in the hormone scheme are required during the transformation procedure with cyanamide selection compared to selection on kanamycin. Upon application of a transformation protocol for Russet Burbank, only in 1 % of the plants (in a population of 241) Agrobacterium could be detected, but these bacteria did not contain the vector for transgene transfer anymore.

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Abbreviations

TE:

Transformation efficiency

EE:

Escape efficiency

D:

2,4-Dichlorophenoxy acetic acid

Z:

Zeatin riboside

G:

Gibberilic acid

I:

Indolacetic acid

IBA:

Indolbutyric acid

NAA:

Naphtylacetic acid

BAP:

Benzylaminopurin

GA:

Gibberillic acid

OD600 :

Optical density at 600 nm

GUS:

Beta-glucuronidase

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Acknowledgments

D. Ingham for the Taqman analysis, D. Palmer for the field evaluation, S. Tito for the PCR analysis, H. Mudjiwarti and A. Kramer for technical support. Furthermore, it should be mentioned that the work presented was done at Syngenta Mogen, which has been disbanded in the year 2002; this manuscript is an homage to all these people involved and especially our colleague Stephan Ohl, who deceased far too early.

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Authors and Affiliations

  1. Syngenta Mogen, Einsteinweg, Leiden, The Netherlands

    Hella van Driel, Marina Keppel, Edwin van den Berg, Walter Schoo, Margaretha van der Lans, Marion Notkamp, Ellis Jansen, Eugene Roebroeck, Stephan Ohl & Sietske Stuiver-Hoekstra

  2. Anthura BV, Anthuriumweg 14, 2665 kV, Bleiswijk, The Netherlands

    Hella van Driel, Walter Schoo & Marion Notkamp

  3. Rijk Zwaan Breeding BV, Eerste kruisweg 9, 4793 RS, Fijnaart, The Netherlands

    Marina Keppel & Edwin van den Berg

  4. Monsanto Vegetable Seeds Division, P.O. Box 1050, 2660 BB, Bergschenhoek, The Netherlands

    Margaretha van der Lans

  5. Lgem, Madepolderweg 45C, 2553 EG, The Hague, The Netherlands

    Eugene Roebroeck

  6. Sigash, An der Althart 31, 67433, Neustadt Weinstrasse, Germany

    Sietske Stuiver-Hoekstra

Authors
  1. Hella van Driel
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  2. Marina Keppel
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  3. Edwin van den Berg
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  4. Walter Schoo
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  5. Margaretha van der Lans
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  7. Ellis Jansen
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  8. Eugene Roebroeck
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  9. Stephan Ohl
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  10. Sietske Stuiver-Hoekstra
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Corresponding author

Correspondence to Sietske Stuiver-Hoekstra.

Additional information

Stephan Ohl: Deceased.

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van Driel, H., Keppel, M., van den Berg, E. et al. Cyanamide hydratase as selectable marker in potato. Plant Cell Tiss Organ Cult 118, 125–135 (2014). https://doi.org/10.1007/s11240-014-0468-6

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  • DOI: https://doi.org/10.1007/s11240-014-0468-6

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