Workpackage 2: Breeding Systems Specific objectives transformation research Development of a reliable transformation protocol for garlic using Agrobacterium tumefaciens as a vector The production of transgenic garlic with an altered S metabolism Persons involved Si-Jun Zheng, Betty Henken, Frans Krens, Chris Kik (Plant RI, Wageningen, the Netherlands)
Short overview of research 2002-2003 In vitro: callus induction and plant regeneration on non-apical parts of roots. Genetic transformation: two transformation systems via GUS and GFP; molecular analysis on transgenic plants. Genetic modification of sulphur metabolism: gene isolation and fusion, transform APS1 gene into garlic.
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Workpackage 2: Breeding Systems
Specific objectives transformation research Development of a reliable transformation protocol for
garlic using Agrobacterium tumefaciens as a vector The production of transgenic garlic with an altered S
metabolism Persons involved
Si-Jun Zheng, Betty Henken, Frans Krens, Chris Kik(Plant RI, Wageningen, the Netherlands)
Short overview of research 2002-2003
In vitro: callus induction and plant regeneration on non-apical parts of roots.
Genetic transformation: two transformation systems via GUS and GFP; molecular analysis on transgenic plants.
Genetic modification of sulphur metabolism: gene isolation and fusion, transform APS1 gene into garlic.
Development of garlic regeneration system
(Zheng et al., 2003. In Vitro Cell. Dev. Biol. Plant 39: 288-292)
Development of a garlic genetic transformation system
GUS transformation system destructive
GFP transformation system non-destructive
Genetic transformation: analysis of transgenic garlic via GUS
Genetic transformation: analysis of transgenic garlic via GFP
Garlic plants with GFP expression after selection for 4 months and regeneration for another 3 months (cv. Printanor)
Analysis of transgenic garlic: standard PCR uid A primers
resulting in a 710 bp fragment
lane 1-3: transgenic garlic
lane 4: negative control
lane 5: positive control lane 6: 1kb DNA
ladder marker 1 2 3 4 5 6
Genomic DNA blot : 802 bp fragment of Cry1Ca PCR
product as a probe lane 1-13: individual transgenic garlic plant transformed with with pCAMBIA1301- Cry1Ca
lane M: DNA digested with Hind III
lane N: untransformed garlic DNA as negative control