Download Production of bright-colour poinsettia using Agrobacterium

Addition of novel-colour to the commercial poinsettia cultivar using
Agrobacterium tumefaciens-mediated genetic transformation
Introduction:
Biotechnology offers new means of improving ornamental plants by the addition of desirable
traits to existing cultivars. Poinsettia (Euphorbia pulcherrima Willd. Ex Klotzsch), is a
contemporary symbol of Christmas in most parts of the world. In Norway, poinsettia is one of
the most important pot plants with a yearly production close to 6 million plants. The annual
production in the USA and the EU is respectively 50 mill and 100 mill plants. Its ornamental
value and innovation potential has laid the basis for an extensive research in Norway. One of
the main focuses is to initiate the creation of novel-coloured poinsettia, a highly desirable trait
for the poinsettia industry through the engineering of aurone synthase pathway by
Agrobacterium-mediated transformation method.
In 1907, Smith and Townsend demonstrated that the Gram-negative soil bacterium
Agrobacterium tumefaciens, can cause crown gall tumors in plants as a result of bacterial
infection, usually at the wound sites (Herrera-Estrella et al. 2004). Later in the 1980s,
scientists in Europe and USA successfully used the T-DNA region to insert foreign genes into
tobacco and showed that foreign genes became integrated and functionally expressed in the
plant cells. Since then, T-DNA region has been largely modified so that the disarmed Ti
plasmids, which contain a T-DNA lacking genes involved in tumor formation, can be
purchased and used for genetic modification of plants. The disarmed plasmid will allow the
Agrobacterium to insert foreign DNA into plant cells without causing crown gall tumours. A
simplied schematic demonstration of Agrobacterium tumefaciens method is shown below
(Adapt from Dr Micheal Parkinson 2002).
The Agrobacterium-mediated transformation method developed at Bioforsk. Application for
patenting the method is filed and the method has been used to generate Poinsettia mosaic
virus (PnMV) resistant poinsettia via somatic embryogenesis (se pictures below).
Project: Research Council of Norway (Norges forskningsråd) project.
Objectives: To generate transgenic poinsettia with yellow colour. Two genes (AmAs1 and
4’CGT) involving in the aurone biosynthesis conferring bright yellow colour are inserted into
T-DNA vectors for transformation.
Milestones:
1. Agrobacterium-mediated transformation of poinsettia (julestjerne) will start at the
beginning of November, 2007.
2. Tissue culture and selection of transformants (November 2007-June 2008).
3. Analysis of transformants (February 2008-June 2008)
4. Results and publishing (Winter 2008).
What a Master student can gain/learn:
1. Tissue culture technique
2. Get an insight into the gene technology and its potentials.
3. Agrobacterium-mediated transformation method
4. Molecular methods such as PCR, Southern and Northen blot analyses.
5. Writing skills and co-author for publication on international-peer-reviewed
journals if the contribution is adequate.
Supervisors: Dr Trine Hoslef-Eide (UMB), Dr Jihong Liu Clarke (Bioforsk) and Dr Dag
Ragnar Blystad (Bioforsk).
Literatures relating t the project and detailed information about the project can obtained by
contacting Dr Trine Hoslef_Eide (XXXX) or Dr Jihong Liu Clarke by address shown below.
Jihong Liu Clarke, PhD
Senior Research Scientist & Coordinator for China collaboration
Department of Genetics and Biotechnology,
Plant Health and Protection Division,
Norwegian Inst for Agricultural & Environmental Research,
Hoegskoleveien 7, N-1432 Aas, Norway
Tel: +47-92609264 (O); +47-99594790 (M)
Fax: +47-64946110; email: [email protected]
www.bioforsk.no