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Tissue and plastid targeted transgene expression in a perennial plant, strawberry - HH1031SSF

This proposal focuses on developing the ways and means of targeting transgene expression in strawberry. Two technologies will be used: 1) plastid transformation utilising a novel, non-antibiotic based approach for selection based on carbohydrate metabolism and 2) nuclear transformation with strawberry tissue-specific promoters fused to a marker gene and determination of precision of promoter activity. Plastid transformation should exclude transgene transmission through pollen. Plastid inheritance will be investigated within cultivated strawberry and also between cultivated and wild strawberry. This strategic research underpins development of genetically modified (GM) strawberry and focuses on safety. It is directly relevant to MAFF’s science strategy for research using genetic modification technology. It aims to reduce potential and perceived risks in relation to the consumer and non-target species, through targeting of transgene expression to only those tissues where the expression is required, and in relation to the environment, through containment of transgenes through non-release via pollen. It also addresses elimination of antibiotic resistance markers through development of a novel non-antibiotic-utilising system for plastid transformation. Data obtained from this work will be published in the relevant scientific journals and communicated to the soft fruit industry and consumers to increase awareness of the benefits of such targeted transgene expression.
This work will provide the molecular tools for application of GM technology in plants aimed at overcoming public concerns surrounding the use of antibiotic resistance genes, pollen transfer and expression of transgenes throughout the whole plant. The scientific objectives for this research are:
1. Development of the xylose isomerase/xylose selection system for plastid transformation in strawberry (Nov. 2000 – Oct. 2002).
2. Tissue culture of plastid transformed lines towards obtaining stable, homoplasmic lines for future inheritance studies (Nov. 2001 – Oct. 2002).
3. Production of strawberry nuclear transformed transgenic plants containing plasmid constructs with floral organ and root-specific promoters fused to the uidA (GUS) gene (Nov. 2000 – April 2002).
4. Analysis of promoter activity in flowering populations of the transgenic plants transformed with tissue-specific promoter constructs (April 2002 – Oct. 2002).
5. Identification of plastid genome sequence markers in strawberry parental genotypes using PCR-RFLP and PCR-SSCP (Nov. 2002 – July 2003).
6. Determination of whether plastids are transmitted through pollen in strawberry using plastid genome sequence markers (August 2003 – March 2004)

Project Documents
• Final Report : Tissue and plastid targeted transgene expression in a perennial plant, strawberry   (3251k)
Time-Scale and Cost
From: 2000

To: 2004

Cost: £393,734
Contractor / Funded Organisations
Horticulture Research International
GM Food              
Fields of Study