The green fluorescent protein as an efficient selection marker for Agrobacterium tumefaciens-mediated transformation in Hevea brasiliensis (Müll. Arg)
Leclercq J., Lardet L., Martin F., Chapuset T., Oliver G., Montoro P.. 2010. Plant Cell Reports, 29 (5) : p. 513-522.
An efficient genetic transformation procedure using a recombinant green fluorescent protein (GFP) has been developed in Hevea brasiliensis clone PB260. Transformation experiments have been performed using an Agrobacterium tumefaciens binary vector harbouring both uidA and S65T-GFP reporter genes in order to compare selection methods using glucuronidase assay (GUS activity) and paromomycin resistance, GFP activity and paromomycin resistance, or GFP activity only. At transient level, the number of spots showing GUS or GFP activities was similar for 4 and 5 days after coculture. After selection, stable transformation events were observed and led to the establishment of transgenic callus lines. A higher number of lines were generated with GFP selection compared to the GUS one. GFP selection is less time-consuming in terms of callus subculturing, and offers the possibility of producing antibiotic resistance marker-free transgenic plants.
Mots-clés : hevea brasiliensis; agrobacterium tumefaciens; transformation génétique; marqueur coloré; fluorescence; gène; cytotoxicité; embryon somatique; marqueur génétique; résistance aux antibiotiques; languedoc-roussillon; france; gus
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Agents Cirad, auteurs de cette publication :
- Leclercq Julie — Bios / UMR AGAP
- Martin Florence — Bios / UMR AGAP
- Montoro Pascal — Bios / UMR AGAP
- Oliver Gérald — Bios / UMR AGAP