Faculty of Biology, University of Latvia
EEB
Hard copy: ISSN 1691–8088
On-line: ISSN 2255–9582
Acta Univ Latv (2004) 676: 223–226
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Environmental and
Experimental
Biology

Acta Univ Latv (2004) 676: 223–226

Orginal Article

A transformation method for obtaining marker-free plants based on phosphomannose isomerase

Karin Sonntag1*, Youping Wang1, Michael Wallbraun2
1Federal Centre for Breeding Research on Cultivated Plants, Institute of Agricultural Crops, Rudolf-Schick-Platz 3a, Gross Luesewitz D-18190, Germany
2Centrum Grüne Gentechnik, DLR-Rheinpfalz, Breitenweg 71, Neustadt an der Weinstraße 67435, Germany
*Corresponding author, E-mail: k.sonntag@bafz.de

Abstract

A selectable marker system for plant transformation of rape plants that does not require the use of antibiotics or herbicides was developed. The selectable marker consists of the phosphomannose isomerase (PMI) gene from E. coli under the control of a plant promoter. The PMI gene was transferred to Brassica napus by Agrobacterium- mediated transformation, which allows the selection of transgenic plants with mannose and sucrose as selective agents. The highest transformation frequency of 7.9 % was obtained when a combination of 4.5 g l-1 mannose and 10 g l-1 sucrose was used. For early identification of transgenic events, histochemical staining with 5-bromo-4-chloro-3-indolyl-ß-D-glucuronide (X-Gluc) was used. Stable integration of the transgene was confirmed by PCR and Southern blot analysis. These preliminary results indicate that the mannose selection system can be used for Agrobacterium-mediated transformation of rape plants.

Key words: Agrobacterium tumefaciens, mannose selection marker, rape, transformation

 
Acta Univ Latv (2004) 676: 223–226
 DOI: http://doi.org/10.22364/eeb
EEB

Editor-in-Chief
Prof. Gederts Ievinsh



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University of Latvia

 
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